Review

Received: 27 March 2010; Accepted: 27 March 2010; Published online in Wiley Online Library: 12 May 2010

(wileyonlinelibrary.com) DOI 10.1002/ffj.1996

Biotechnology for the production of essential

oils, flavours and volatile isolates. A review.†,‡
Y. Gounaris*

ABSTRACT: Various applications of biotechnological methods for the production of volatile compounds useful to the food and
pharmaceutical industries are discussed. The yields obtained from intact or genetically modified plants are compared to those
achieved by microbial methods. Plant yields are too low for the products to compete commercially to those synthesized
chemically. Still lower yields are obtained with in vitro-cultured plant tissues. Trangenic plants with altered methylerythritol
path gave 50% more essential oil in the best case. The 100-fold increases in shikimate-derived volatiles, obtained with over-
expressed alcohol dehydrogenase and five-fold more C6 volatle aldehydes and 2-phenylethanol, were produced with overex-
pressed lipoxygenase and 2-phenylethanol dehydrogenase, respectively. However, the most spectacular yields were observed
with biotransformations catalysed by microorganisms. Kluyveromyces marxianus, produces over 26 g/l 2-phenylethanol from
phenyalanine, whereas Candida sorbophila, Mucor circillenoides or Yarrowia lipolytica can produce 5–40 g/l g-decalactone from
ricinoleic acid. Vanillin production from ferulic acid is in the range 12–60 g/l with Amycolatopsis and Streptomyces species.
Vanillin can be produced at 5 g/l by Escherichia coli and amorphadiene yields of 37 g/l have been observed with Saccharomyces
cerevisiae, both with the genetically overexpressed methyl–erythritol path. Genetically engineered b-oxidation genes result in
yields of 10 g/l g-decalactone byYarrowia lipolytica and up to 80 g/l dicaboxylic acids by various yeasts. These results far exceed
the theoretical limit of about 1 g/l required for consideration of a procedure as a commercially interesting process, alternative
to chemical sythesis. Copyright © 2010 John Wiley & Sons, Ltd.

Keywords: bioreactor; biotechnology; essential oil; terpenoids; volatiles

Introduction plant tissues in limited quantities. Plant seeds, flowers, stems

There are hundreds of thousands of different secondary metabo-
lites produced by plants, four times more than the number pro-
duced by microorganisms. This number is estimated to represent
only 10% of the secondary metabolites existing in plants and still
waiting to be isolated and identified. Of these, terpenoids com-
prise the largest and structurally most varied class, numbering
over 40 000 different molecules. Members of the 10-carbon ter-
penoids, the monoterpenoids, are constituents of essential oils
produced by plants. The essential oil monoterpenoids are vola-
tile, which means that they pass in to the air in sufficient concen-
trations to be detected by, and to act on, other organisms.
Essential oils can also contain sesquiterpenoids, phenypro-
panoids and benzenoids. In addition, plant tissues can produce
volatile aldehydes and their corresponding alcohols, and acids as
well as volatile ketones. These compounds are occasionally found
in essential oils, but are usually formed in specific plant tissues
and under specific physiological conditions that favour catabolic
reactions. They can be considered as belonging to the primary
metabolism, although they can have useful fragrance, flavour or
medicinal qualities.
The commercial interest on volatiles stems from their aro-
matic and flavour qualities. Several of them, have significant
antimicrobial and antineoplastic activity. Others act as messen-
gers in communication between plants themselves or with
other organisms. Volatiles are obtained from plants by distilla-
tion at or by extraction with ethanol, diethyl ether, chloroform,
pentane, hexane, benzene or other organic solvents. Unfortu-
nately, volatiles, like most secondary metabolites, are present in
and roots usually contain 0.1–10% v/w fresh weight essential oil
and often <0.1%. Cases of up to 20% v/w or even higher essen-
tial oil concentrations in plant tissues are also known. Even so, a
single compound can constitute 40–90% of the oil and usually
is not the most useful one. The most desirable volatiles are often
present in the essential oil at concentrations <10% and even
less than 1%. Considering that in most cases the essential oil is
produced in only specific plant tissues, such as seeds or flowers,
whose total mass in a single harvesting season is a small weight
percentage of the whole plant, it is obvious why obtaining
useful volatiles from cultivated plants can be an expensive
operation. Chemical manufacture of small organics, such as the
majority of plant volatiles, is often cheaper, so that the natural
product occupies a small percentage of the market. An example
is vanillin, nine-tenths of whose market involves the synthetic
product.[1] A factor determining the share market of the ‘natural’
* Correspondence to: Y. Gounaris, 10 Stoimenidou Street, 62045 Alistrati,
Greece. E-mail: igouna@otenet.gr
†
This article is part of the Special Issue of Flavour and Fragrance Journal
entitled, Aromatic Plants, Spices and Volatiles in Food and Beverages™, edited
by Ana Cristina Figueiredo and M. Graça Miguel
University of Thessaly, Department of Agriculture, Fytokou Street, 38446 New
Ionia, Magnesia, Greece
‡
This article was published online on 12 May 2010. The funding information in
this footnote has been removed. This notice is included in the online and print
versions to indicate that both have been corrected 28 May 2010
367

Flavour Fragr. J. 2010, 25, 367–386 Copyright © 2010 John Wiley & Sons, Ltd.

product compared to that of the synthetic is the recent
preference of consumers for it, partially stemming from the
belief that it is free from traces of harmful manufacturing arte-
facts and left-overs. Also, the chemical synthesis often results in
racemic mixtures of the product, giving an approximation only
of the natural flavour qualities. There is a need to reduce the
cost of the natural product, so it becomes available to a wider
range of consumers. From an environmental aspect, the produc-
tion of useful volatiles by non-chemical environmentally
friendly methods is always a preferred and often necessary
alternative.
Biotechnology attempts to facilitate the production, and
therefore to reduce the market cost, of natural volatiles by
employing a variety of non-polluting methods. The initial
attempts with plant materials consisted of efforts to produce
volatiles in plant cell or tissue cultures, either by de novo synthesis
or by biotransformation of cheap precursors into high-value
products. Before that, biotransformation efforts involved fungi,
yeast and bacterial cultures. Semi-synthetic methods, in which a
precursor is transformed into a useful product by isolated
enzyme preparations, crude or purified, have also been
attempted. Recently, most efforts have involved metabolic engi-
neering of the biosynthetic pathways leading to the synthesis of
the desired volatile. The various methods of biotechnology for
producing useful volatiles are discussed in this paper.
Types of Volatile Metabolites with Flavour
and Fragrance Qualities or with Biological
Action and Their Biosynthesis
Of the terpenoids, only members of the mono- (C10) and ses-
quiterpenoid (C15) classes are sufficiently volatile. Volatility is
determined not only by the size of the molecule and its stere-
ochemistry, but mainly by its ability to form hydrogen bonds.
Monoterpenoids whose molecule contains only carbons and
hydrogens are very volatile. Those with one hydroxyl, keto,
peroxy, or epoxy group are still volatile, but those with more
hydroxyls are only slightly or not at all volatile. The same holds
for the sesquiterpenoids, where one hydroxyl group seems to
be the maximum tolerated hydrogen bond-forming function for
sufficient volatility to be preserved in the molecule. Triterpe-
noids and higher-order terpenoids are not volatile. Phenylpro-
panoids and benzenoids, bearing up to one hydroxyl and no
carboxyl, are volatile. The simultaneous presence of a keto
group does not abolish the volatility. However, more hydroxyls
drastically reduce or completely abolish the volatile properties.
(Hydroxy)cinnamic acids are not sufficiently volatile, due to the
presence of the carboxyl group, unless it is esterified with a
volatile alcohol. Aliphatic and olefinic aldehydes, monoalcohols
and monoketones are volatile for at least up to 12-carbon sizes.
As the molecule becomes smaller than five carbons, even the
acids are volatile. Organic monocarboxylic acids, esterified with
volatile alcohols, are also volatile. This type of compounds can
be divided in two categories. One has linear carbon chains and
the other has a methyl side-chain.
Genetic engineering is a powerful method used to alter the
rate of volatile production by acting on the biosynthetic path-
ways leading volatile synthesis. A short discussion of these path-
ways seems pertinent at this point. The monoterpenoids are
produced by the plastidic methyl–erythritol–phosphate (MEP)
368
path, whose sequence and enzymatic properties have been
View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd.
Y. Gounaris
elucidated almost to completion.[2–9] Its rate-limiting step is the
one catalysed by 1-deoxy-D-xylulose 5-phosphate synthase
(DXS). NADPH, CTP and ATP are required for its operation. Con-
densation of dimethylallyl diphosphate (DMADP) with isopente-
nyl diphosphate (IDP) by the action of geranyl diphosphate
(GDP) synthase leads to formation of the monoterpenoid pre-
cursor GDP. Like all prenyltransferases, the GDP synthase is a
rather slow catalyst. The cis isomer neryl diphosphate (NDP) is
also formed. Cyclization of GDP is catalysed by the also slow
cyclases, membrane-bound enzymes in the plastids and endo-
plasmic reticulum. The hydroxylations of linear or cyclic monot-
erpenes are catalysed by NADPH-consuming, cyt450-dependent
monoxygenases, utilizing molecular oxygen, but also able to use
hydrogen peroxide produced from any source. These hydroxy-
lases are inducible by a variety of biotic or abiotic stress factors.
Sesquiterpenoids are considered to be synthesized in the
cytosol from farnesyl diphosphate (FDP), derived from the
mevalonic acid pathway. Two NADPH and three ATP molecules
are consumed for FDP synthesis and the rate-limiting step is
catalysed by 3-hydroxymethylglutaryl coenzyme A reductase
(HMGR). Sesquiterpene cyclases act on FDP to produce at least
200 types of cyclic sesquiterpenoids.
The phenylpropanoids are produced from phenylalanine and
tyrosine, both derived from the shikimic acid pathway. A multi-
tude of feedback-inhibited steps and a need for NADPH and ATP
in the shikimic acid path ensure a tight regulation of Tyr and Phe
synthesis. The requirement for NADPH is even greater in the
transformation of Phe and Tyr into phenylpropanoids. It is
required for the removal of the carboxyl group of the propenyl
side-chain by successive reductions that form the corresponding
volatile aldehydes, alcohols and phenylpropenes. The carboxyl
group is first esterified to coenzyme A, by specific ligases. Then it
is transformed to an aldehyde group by an oxidoreductase and
the aldehyde is reduced into an alcohol by an alcohol dehydro-
genase (ADH). Phenylpropenes are then produced from the
alcohols. NADPH is also required for the hydroxylations of the
aromatic ring. Benzoic and phenolic acids come from the corre-
sponding hydroxycinnamic acids by b-oxidation of the propenyl
chain, followed by oxidative decarboxylation. This process is
tightly regulated by feedback inhibition. Volatile derivatives
are then formed by the reduction of the carboxyl group, as in
phenylpropanoids.
Non-branched volatile aldehydes and their corresponding
alcohols can be derived by degradation of unsaturated fatty
acids, mainly linoleic and linolenic acid,[10–12] by the sequential
action of lipoxygenases (LOX), hydroperoxide lyases (HPL) and
ADH. The initial introduction of molecular oxygen into the
carbon–carbon double bonds requires NADPH. These are cata-
bolic reactions of the primary metabolism. Methyl-branched
compounds, such as isovaleric and isobutyric acid, are derived
from leucine and valine catabolism. Isovaleric acid could poten-
tially be produced from DMADP of the terpenoid synthesis
pathways. According to Höschle et al.,[13] branched volatiles can
be produced by catabolism of lineal terpenoids and of leucine-
derived 3-methyl-crotonyl-CoA, the precursor of isovaleric acid,
via degradation of the produced 3-methyl glutaryl-CoA. In bac-
teria at least, they could be produced from linear monoterpe-
noid or even sesquiterpenoid degradation.[14,15] Unlike the LOX
path, the degradations of leucine, valine, monoterpenoids and
DMADP, leading to volatile aldehyde, alcohol and acid produc-
tion, do not need reductive equivalents but rather produce
them.
Flavour Fragr. J. 2010, 25, 367–386
Biotechnology for essential oils, flavours and volatile isolates
De novo Production of Volatiles by Tissue
and Cell Cultures
Plant Tissue and Cell Cultures
Volatiles in callus and cell cultures. Producing compounds in
plant cell, callus or tissue cultures has been attempted to ensure
a stable supply and quality of the product. Some of the plants
used as sources for the desired substance are rare, slow-growing
and found in not easily approachable regions of the world and
difficult to cultivate. The in vitro cultures were expected to speed
up the biomass propagation rate and to have it under controlled
conditions and immediately available. These attempts encoun-
tered serious difficulties. The rate of secondary metabolite pro-
duction by in vitro-cultured plant cells is orders of magnitude
lower than that in the intact plant, usually in the range 0.1–
0.01 g/l day.[16] Volatile compound yields are still lower and vola-
tile secondary metabolites are present often in trace amounts
detected in cultures of various plant species, examples of which
are given in Table 1. Although cases of cultures showing higher
yields of secondary metabolites than the intact plant are
known,[41] they are not involving volatiles. In many cases, the
volatiles found in the intact plant are not present at all in the in
vitro cultures. They are often different than those present in the
intact plant. Volatile aldehydes, alcohols, ketones and acid esters
appear more frequently or for the first time in in vitro cultures. The
terpenoids produced are in most cases glycosylated.
The reasons for the reduced ability of the in vitro cultures
to produce volatiles, and secondary metabolites in general,
are not known with certainty. The cultured cells and callus seem
to have some enzymatic activity for terpenoid production.[42–44]
Geranyl diphosphate synthase activity has been detected in
plastids[45] but sesquiterpene cyclase has not.[46–48] According
to Falk et al.,[49] the inability of cultured plant cells and callus
to accumulate significant amounts of monoterpenes, could be
due to the combined effect of lower enzymatic activity and
their higher catabolic rate. Concerning the phenylpropanoid
synthesis potential, enzymatic activities of phenylalanine
ammonia lyase, shikimate dehydrogenase, cinnamic acid-4-
hydroxylase, p-coumaric acid-3-hydroxylase, cinnamoyl-CoA
reductase, 4-coumarate:CoA ligase, 4-hydroxycinnamate:CoA
ligase, cinnamyl alcohol dehydrogenase and caffeic acid-O-
methyltransferase have been detected in callus or cell suspen-
sions and are often equal to those in the intact plant.[50–54] Of the
enzymes of the volatile aldehyde and alcohol synthesis path,
discussed below, lipoxygenase and hydroperoxide lyase activity
has been found to be present in in vitro-cultured plant tis-
sues.[29,55,56] In cell suspension cultures of alfalfa, the hydroper-
oxide lyase activity was rate-limiting.[57] The ability of cultured
plant tissue and cells to produce volatiles is inducible by a
variety of chemical and physical factors, as is also the ability for
secondary metabolite synthesis in general. The induction treat-
ments increase the essential oil yield by up to five-fold[58,59] in
some oils containing novel terpenes or in oils of altered relative
percentage.[60–62] Even under the optimum induction conditions,
the yield of essential oil by in vitro-cultured plant tissues and
cells is usually less than that achieved by the intact untreated
plant. Therefore, using cultured plant cells and calli for volatile
production, even with the inclusion of elicitors and other
inducers, does not seem to be a particularly promising
undertaking.
Flavour Fragr. J. 2010, 25, 367–386 Copyright © 2010 John Wiley & Sons, Ltd.
Volatiles in hairy roots. It is a general observation that sec-
ondary metabolite yield by cell and callus cultures increases if
some degree of cell differentiation is induced. Genetic transfor-
mation of plant tissue by insertion of the T-DNA regions of the Ri
plasmid of Agrobacterium rhizogenes results in the formation of
small, fine, hair-like root structures, known as ‘hairy roots’. Four
of the 18 ORFs in the TL-DNA are essential for hairy root formation,
of which ORF11 (rolB) is absolutely necessary. The TR-DNA carries
two auxin synthesis genes, but by itself does not provoke hairy
roots formation. Hairy roots lack geotropism, are highly branched
and can be cultured in bioreactor facilities needing no plant
growth regulators, since the inserted T-DNA carries genes for
auxin synthesis. They grow as fast, or faster, than normal roots,
with meristem cell cycles averaging 10 h. They produce second-
ary metabolites at levels and patterns similar to those of normal
roots, but also metabolites produced in aerial parts of the plant.
Often novel compounds are also produced. Unlike cell or callus
cultures, hairy roots are biochemically stable and the T-DNA is
stably integrated.
Excellent reviews on the culture methodologies and morpho-
logical and biochemical characteristics of hairy root cultures,
including their potential for secondary metabolite production,
have been published.[42,63–66] Most of these reviews cover the wide
spectrum of secondary metabolites and the preponderance of
the cited cases concerns the production of non-volatile com-
pounds, primarily alkaloids and secondarily some phenolics and
non-volatile higher terpenoids, with a few cases involving vola-
tiles. However, Figueiredo et al.[63] focused on essential oils only;
they listed 11 plant species whose hairy root cultures can synthe-
size essential oil constituents. Among these, Pimpinella anisum
and Achillea millefolium hairy roots are capable of essential oil
yields similar to, or even higher than, those obtained with the
roots of the parent plants. It is clear that hairy roots have the
potential for synthesizing both volatile and non-volatile
terpenoids.
Table 2 presents a list of additional examples, specifically for
volatile compounds produced by hairy roots, mostly drawn from
Figueiredo et al.[63] In these, the essential oil was analysed to some
extent, although the authors also cite the cases of Daucus carota
and Leontopodium alpinum hairy roots, whose main component
was not identified. The yields are greatly elevated in comparison
to those of cell or callus cultures and can be further increased by
the inclusion of abiotic or biotic elicitors in the culture medium. A
prospect for further increasing the volatile production from hairy
roots is to genetically engineer their volatile production paths
using transgenes inserted into the T-DNA region.
Volatile Synthesis by Cultured Microorganisms
Although a great deal of research on the biotechnology of vola-
tiles still involves plants, especially efforts to increase terpenoid
and phenolics production in transgenic plants, most of the recent
effort is directed to using microorganisms. Volatile aldehydes and
alcohols are far more easily produced by cultured microorgan-
isms, and efforts to genetically alter microbes for producing or
biotransforming terpenoids or phenolics were met with reward-
ing success. Therefore, in most cases microorganisms are used for
their production, instead of plant cell cultures. Also, microorgan-
isms (bacteria, algae and fungi, including yeasts) are sturdier than
plant cells under bioreactor conditions. They are better suited to
withstand the frictional stress imposed by the shaking proce-
369
dures as well as various temporary extremes of pH, temperature
View this article online at wileyonlinelibrary.com
 370

Table 1. Volatiles detected in plant cell or callus cultures

Plant Products/remarks Reference

Agastache rogosa (Korean mint) Volatile C9-aldehydes and alcohols, butanedione. Different from those in intact plants [17]
Artemisia dracunculus (tarragon) Production of phenylpropenes of the essential oil [18]
Citrus sp. C. paradisi callus produced 40 volatiles (mono-, sesquiterpenes, adehydes and hydrocarbons), 186 mg/kg FW. This is 5% of [19]
peel oil yield. C. limon produced 11 monoterpenes and n-nonanal, 40 mg/kg FW. C. aurantifolia gave only limonene,

View this article online at wileyonlinelibrary.com

4.4 mg/kg FW
Citrus sinensis No volatile components were detected, but embryogenic callus produced 10 ingredients of orange oil [20]
Citrus aurantifolia Produced citrals, terpenyl acetate, dodecanal [21]
Coleonema album Monoterpenes. More if cultures under light [22]
Eucalyptus camaldulensis Alkanes, alkenes, alcohols in calli derived from stamens [23]
Eucalyptus citriodora Monoterpenes in calli derived from immature flowers [24]
Melissa officinalis (balm) Low amounts of 2-phenylethanol, d-octalactone [25]
Very low amounts of C6-aldehydes, -alcohols and -acetate asters. Large concentrations of glycosides of nerol, citronellol, [26]
geraniol, 1-octen-3-ol
Mentha piperita (peppermint) Mint oil components [27]
Ocimum basilicum Essential oil ingredients [28]
Olea europaea (olive tree) Produce most of the volatile C6-aldehydes, -alcohols and -acetyl ester found also in olive oil [29]
Origanum acutidents Origanum oil ingredients (38) [30]
Origanum vulgare Formation of n-alkanes. Lack of terpenoids even in green calli [31]
Oryza sativa Volatile hydrocarbons, alcohols, ketones aldehydes, esters. Most were present in the intact plant also [32]

Copyright © 2010 John Wiley & Sons, Ltd.

Petroselinum crispum (parsley) Both types of cultures produced nonanal and decanal. Cell cultures produced also limonene, acetophenone, not found in [33]
callus or in intact plants. No phellandrene, apiole, menthatriene, that as found in intact plants
Salvia officinalis (sage) Low amounts of essential oil [34]
Smyrnium perfoliatum a-Pinene [35]
Fragaria sp. (strawberry) Low amounts of ethyl butyrate, butyl butyrate [36]
1,2-Propanediol (flavour precursor) [37]
Tacoma sambucofolium Accumulation of phenylpropanoid glycosides. [38]
Taraxacum officinale (dandelion) Acetate butyl ester, 2-methyl-1-propanol, n-butanol, 4-phenyl-1-butanol, terpineols, 4-hydroxy-4-methyl-2-pentanone, [39]
acetate
Vanilla planifolia Vanillin [40]
Y. Gounaris

Flavour Fragr. J. 2010, 25, 367–386

Biotechnology for essential oils, flavours and volatile isolates

and salt or deleterious metabolite concentrations created during

Reference
the culture process. All classes of volatiles can be produced by

[77]
[67]

[76]
[68]

[70]
[69]

[71]
[72]
[73]
[74]

[75]
microorganisms (Table 3), but aldehydes, alcohols and organic
acid esters are in far greater preponderance, indicating a strong
participation of catabolic processes. Microorganisms producing
volatiles have also been previously cited in several review

2,3-Dihydro-2,6-dimethyl-4H-benzopyran-4-one in the essential oil of hairy and normal roots. Similar constituency of the oils
Essential oil yield was 3–10% of that of normal roots. Contained enhanced percentage of falcarinol, farnesene, phellandrene,
articles.[93,118–124] In most cases the yield of the main compound is
<100 mg/l. Far better yields are attained when a substrate bio-
chemically more immediate to the volatile product is used, or
Essential oil yield was only 0.02%, compared to 0.06%, 0.3% and 2% of normal plant root, leaf and fruits, respectively

when an immediate precursor is included in the culture medium

together with the crude substate, a case closer to the biotrans-

Hairy root oil had kessyl alcohol and kessyl acetate instead of bornyl acetate and valerenal of the normal roots
formation procedures discussed in the next section. For example,
2-phenylethanol production by Kluyveromyces and other yeasts
can exceed 400 mg/l.[91,112] 2-Phenylethanol is a rose-like aroma

3-Hydroxy-4,5-dimethyl-2-furanone (sotolone), 3-amino-4,5-dimethyl-2-furanone (sotolone precursor)

with an annual market of 7000 tons. Chemically produced, is 250
Low amounts of essential oil, containing mainly farnesene. Had also selinene as a new compound

times cheaper than its natural counterpart.[91] High yields of

Essential oil of roots was similar to that of normal roots, with only minor quantitative differences

benzaldehyde (almost 10 g/l) were also achieved by cultured

Essential oil containing 47% neryl isovalerate (precursor of a-fenchene) and 6% neryl butyrate

Goetrichum candidum.[88] Production of the peach-like aroma

g-decalactone by Yarrowia lipolytica and various Candida species
can exceed 10 g/l if castor oil is included in the culture.[81,116]

Biotransformations

Biotransformations by Plant and Microorganism Cultures

Not all steps leading to the synthesis of a volatile are blocked to
the same extent in cultured plant cells and calli. Included in the
culture media, an intermediate compound of the path can often
be (bio)transformed or (bio)converted into volatile products by
Yield of essential oil comparable to that of normal plant

residual enzymatic activity downstream of the blocked step(s).

Eugenol yield was 10-fold lower than in normal roots

Such biotransformation procedures are most successfully used

with microorganisms cultured in supplemented media. The
yields are far better than those in media not supplemented with
a close precursor molecule. Ideally, to keep the cost low and the
process economically worthwhile, cheap abundant precursors
are sought, able to biotransform into high-value products. The
issue of secondary metabolite bioconversions has been devel-
oped in several recent reviews[41,78,93,118–127] and the discussion
elemene, octanal, heptanal
2-Hexenal as main ingredient

here is limited to volatiles only. Some examples are given in

Table 4.
A common problem in biotransformations involving volatiles is
Table 2. Volatiles produced by plant hairy root cultures

that a substrate might not be dissolved easily in aqueous medium.

Product/remarks

Agents facilitating its dissolution can be used, but it must be borne

in mind that above a threshold concentration the substrate might
hinder cell growth. The same holds for the volatile product, which
above a threshold concentration could feedback-inhibit its own
synthesis or could be poisonous to the cells. These practical prob-
lems limit productivity. Methods for continuous addition of
substrate at non-harmful concentrations and removal of the prod-
uct(s) have evolved. Among these, the pervaporation method[230]
Chamomila recutita (chamomile)

and the inclusion of volatile-binding materials in the culture

Levisticum officinale (lovage)

Trigonella foenum-graecum

media are commonly used. The issue of sequestering the product

Anethum graveolens (dill)

has been discussed by Ramachandra and Ravishankar[41] and

Leontopodium alpinum

Georgiev et al.[65] Addition to the culture media of a non-soluble

Artemisia absinthium

Valeriana officinalis
Pimpinella anisum

solid or non-miscible hydrophobic liquid phase creates ‘two-

Ambrosia trifida

Coluria geoides

phase systems’, consisting of two immiscible liquids or of a solid

Cucumis melo

and a liquid phase, in which system the desired volatile is seques-

tered by binding non-covalently to the solid or to the hydrophobic
liquid phase. For example, including the Amberlite resin XAD-4 in
Plant

the cell cultures of Vanilla fragrans increased the yield of vanilla

371

flavour. Alternatively, the product can be bound non-covalently to

Flavour Fragr. J. 2010, 25, 367–386 Copyright © 2010 John Wiley & Sons, Ltd. View this article online at wileyonlinelibrary.com
 372

Table 3. Volatile compounds produced by cultured microorganisms

Microorganism Substrate/culture type Product/remarks Reference

Acetobacter sp. Fusel oil Methylbutyric acid (precursor to aromas) [78]
Aspergillus niger Rice bran oil (4 g/l ferulic acid) 2.8 g/l Vanillin [79]
Coconut fat 2-Undecanone, 2-nonanone, 2-heptanone. 40% yield [80]
Aspergillus oryzae castor oil g-Decalactone, 0.86 g/l [81]
Botryodiplodia theobromae Jasmonic acid, 1100 mg/l (100 mg/g dry cells) [82]
Candida guilliermondii and other Castor oil, decanoic acid g-Decalactone, up to 10 g/l with castor oil hydrolysate [81]
Candida species
Ceratocystis fimbriata Coffee husks / Solid-state 12 Volatiles, including ethanol, acetaldehyde, ethyl acetate (main component, 250 mg/l per kg dry substrate), ethyl [83]
propionate, isoamyl acetate
Pervaporation bioreactor Esters and alcohols. Banana-like flavour [84]
Ceratocystis moniliformis Pervaporation bioreactor Ethyl-, propyl-, isobutyl- and isoamyl-acetates, citronellol, geraniol. All <100 mg/l in the culture broth, but pervaporation [85]
increased total yield by 50% (citronellol) to 414% (isobutyl acetate)
Coryne-bacterium glutamicum Pyrazines (roasted nutty flavour) [78,86]
Fistulina hepatica Oak wood powder 53 Volatile compounds (aldehydes, short-chain alcohols, terpenes, methoxybenzenoids). Mushroom aroma [87]

View this article online at wileyonlinelibrary.com

Geotrichum candidum Leucine+ ethanol 2-Hexanoic acid (9.5 g/l), benzaldehyde (1.6 g/l), acetate asters, methyl-butanol, methyl-propanol, butyric acid [88]
Geotrichum fragrance Leucine+ ethanol Ethyl-isovalerate [78]
Geotrichum klebahnii Leucine+ Ethanol Ethyl esters of branched acids [89]
Castor oil g-Decalactone, 0.2 g/l [81]
Kluyveromyces lactis Grape must + Phe Citronellol, geraniol, linalool [90]
Kluyveromyces marxianus Grape must + Phe 2-phenylethanol, 0.4 g/l [91]
Cassava bagasse, palm bran. Ethyl acetate, ethanol, acetaldehyde, other volatiles [92]
Solid state fermentation.
Moniliella suaveolens Castor oil press cake g-Decalactone,180 mg/kg dry matter [93]
Penicillium chrysogenum Hydrolysed maize fibre Vanillic acid 0.81 g/l, vanillin 0.35 g/l, [94]
Phlebia radiata (Basidiomycetes) Liquid cultures Bisabolol, aromatic and sesquiterpene alcohols. 2-Methyl- and phenyl-propanol were the main ones at 5 mg/l [95]
Piptoporus soliniensis (Basidiomycetes) Liquid media of yeast extract and glucose. g-Decalactone, 8 mg/l. [96]
Polyporus tuberaster K2606 Liquid culture 14 Volatiles, mainly benzylaldehyde (8 mg/l) and 3-methyl-1-butanol [97]
Poriales sp. (Basidiomycetes) Liquid cultures Over 80 volatiles. Mainly bisabolol, geraniol, nerolidol [98]
Proteus vulgaris Cheese Methylbutanals (5 mg/l), methylbytanols (100 mg/l) [99]
Rhizopus oryzae Cassava bagasse with soyabean. Acetaldehyde, 3-methylbutanol, other volatiles [100]
Solid state fermentation
Rhodotorula aurantiaca Castor oil g-Decalactone, 6.6 g/l [101]
Saccharomyces cerevisiae Wine must 2-Phenylethanol, ethyl esters, acetates, volatile fatty acids. Fruity aroma [102]

Copyright © 2010 John Wiley & Sons, Ltd.

Elemental sulphur in the medium Sulphur-containing flavours [103]
Immobilized Alcohols, carbonyls and esters, not significantly different from those of liquid fermentations [104]
Carotenoids Apocarotenoid aldehydes and ketones. Total <4 mg/l [105]
Dough Isoamyl alcohol, phenylethanol [106]
Sporidiolobus salmonicolor Vinasse + ricinoleic acid methyl ester g-Decalactone [107]
Sporobolomyces odorous Hydrolysed castor oil g-Decalactone, 5.5 g/l [108]
Staphylococcus xylosus Starter culture 2- and 3-Methylbutanal (0.8 mg/l), 2-methylpropanal, and many other aldehydes, ketones, esters [109]
Stigmatella aurantiaca Agar cultures C10 volatile ketones [110]
Trichoderma harzianum 6-Pentyl-a-pyrone. Coconut-like flavour [111]
Trichoderma viridae 6-Pentyl-a-pyrone. Coconut-like flavour [112]
Trichoderma harzianum Castor oil g-Decalactone, 260 mg/l [113]
Various other Trichoderma sp. Sugar cane bagasse. Solid state fermentation 6-Pentyl-a-pyrone, 3 g/kg dry matter [114]
Tyromyces sambuceus g-Decanolide, 880 mg/l [115]
Various yeasts Molasses + Phe 2-Phenyl ethanol, 0.9 g/l [112]
Yarrowia lipolytica Castor oil or methyl ester of ricinoleic acid g-Decalactone, up to 12.3 g/l [116]
10% v/v Castor oil or 0.05% decanoic acid g-Decalactone, up to 6.9 g/l [81]
Williopsis saturnus Fusel oil 3-Methyl-butyl-, ethyl-, 2-methyl-butyl-, isobutyl-acetate [117]

Yields are cited if they are referred to explicitly, as many of them are only cited as percentages of GC chromatogram peaks or simply as detected in headspace analyses. Specific substrates are cited if different from the usual culture
media.
Y. Gounaris

Flavour Fragr. J. 2010, 25, 367–386

 Table 4. Biotransformation of precursors for production of volatiles by plants and microorganisms

Organism Substrate Product/effect/yield Reference

Plants
Anethum graveolens Geraniol Nerol was detected for 24 h. Afterwards, no nerol or geraniol was detected [128]
Asparagus officinalis Santonin (sesquiterpene), 0.2 g/l Hydroxylated derivatives, <120 mg/l [129]
Capsicum annuum Ferulic acid, 2.5 mM Vanillin, 10 mg/l [130]
Capsicum frutescens, immobilized cells Coumaric acid, protocatechuic aldehyde, caffeic acid Vanillin, capsaicin [131]
Ferulic acid, vanillylamine Vanillin, capsaicin [132]
Caragana chamlagu Ionones, 0.6 g/l Oxo-, epoxy-, hydroxyl-ionones, epoxyketone (70 mg/l) [133]

Flavour Fragr. J. 2010, 25, 367–386

Catharanthus roseus 3-Carene, 2-pinene Epoxy- and hydroxyl-derivatives [134]
Geraniol, nerol, carvone, 0.1 g/l Hydroxylation at allylic positions, and reduction of double bonds and keto groups, <60 mg/l each [135]
Geraniol Geranial, neral [136]
3-Carene, 2-pinene Epoxy and hydroxyl-derivatives [134]
Hydroxybenzyl alcohols, 60 mg/l Glycosylated derivatives, <20 mg/l each [137]
Piperitone Hydroxylated derivatives [138]
Citrus paradisi, callus Valencene, 5 mg/l Nootkatone, 1 mg/l [139]
[19]
Coffea arabica Vanillin Vanillin glycoside [140]
Dioscorea deltoidea Limonene, 1.2 g/l. Carvone (240 mg/l), via carveol [141]
Eucalyptus perriniana Borneol, 0.5 g/l Borneol glycosides, the most abundant at 240 mg/l [142]
Eugenol, thymol, carvacrol, 0.8 mM Glycosides of thymol, carvacrol and eugenol accumulated in the cells, 70–99% conversion [143]
Eugenol, isoeugenol, 0.5 g/l Glycosides of eugenol and isoeugenol, the most abundant (100 mg/l) [144]
Biotechnology for essential oils, flavours and volatile isolates

Camphor (0.5 g/l) Glycosides of hydroxybornanones from camphor (160 mg/l) [145]
Caryophyllene oxide Caryophylla-3-ene-5,14-diol-O-gentobioside from caryophyllene oxide [145]
Fenchone Glycosylated hydroxyfenchanones [146]
Euphorbia characias Geraniol Geranial, neral [136]
Fragaria sp. (strawberry) a-Ketovalerate Butanal, butanol [36]
Glycyrrhiza alba Monoterpene aldehydes Reduction into alcohols [147]
Lonicera japonica Germacrone Guaiane, eudesmane, secoguaiane [148]
Melissa officinalis (balm) Monoterpenes Metabolize rapidly and disappear [25]
Mentha canadensis Menthyl acetate Menthol that was glycosylated in 24 h [149]
Mentha piperita Menthyl acetate Menthol that was glycosylated in 24 h [149]
Isopiperitenone, 0.1–0.2 g/l Epoxy and hydroxyl-derivatives of isopiperitenone, some of them glycosylated. Also, low amounts of menthol. [150]

Copyright © 2010 John Wiley & Sons, Ltd.

25–35% conversion. All <3 mg/l
Mentha sp., immobilized cells Menthone Neomenthol [151]
Myrtillocactus geometrizans Carene, 0.1–0.5 g/l Carenols (up to 400 mg/l), carenones (100 mg/l) [152]
Nicotiana tabacum 3-Carene, 2-pinene Epoxy- and hydroxyl-derivatives [134]
Hydroxybenzyl alcohols, 60 mg/l Glycosylated derivatives, <20 mg/l each [137]
Geraniol Geranial, neral [136]
Nicotiana tabacum, calli Carene, 0.1–0.5 g/l Carenols (up to 400 mg/l), carenones (100 mg/l) [152]
Nicotiana tabacum, immobilized cells Ionones, 0.1 g/l Corresponding saturated ketones and alcohols, <60 mg/l each [153]
Olea europaea (olive tree) callus Linoleic acid, linolenic acid Hexanal (from linoleic acid), hexenal (from linolenic acid) [29]
Papaver bracteatum Linalyl acetate Linalool, geraniol, a-terpineol [154]
Peganum harmala Geranylacetate, linalyl acetate, 5 mg/l Geraniol (55 mg/l) from geranyl acetate, linalool (14 mg/l) and a-terpineol (12 mg/l) from linalyl acetate [155]
Benzyl acetate, Benzyl alcohol, [156]
anisaldehyde, anisyl alcohol (<80 mg/l),
ionones, ionol, hydro-ionone,
pinenes, verbenol, verbenone, myrtenol, myrtenal,
menthone, menthol, piperitol,
citronellal, <0.1 g/l citronellol
Perilla frutescens Leucine Pinene [157]
Petroselinum crispum Citral, citronellal, geraniol Citral was reduced to geraniol and nerol and citronellal was reduced to citronellol. Geraniol was converted to [158]
nerol, neral and geranial

View this article online at wileyonlinelibrary.com

373
 374

Table 4. Continued

Organism Substrate Product/effect/yield Reference

Picea abies b-Pinene, limonene, 200–400 ml/l From b-pinene: cis- and trans-pinocarveol, myrtenol, a-terpineol, pinocarvone, myrtenal. From limonene: [159]
limonene epoxide, carveol, cineole, perillyl alcohol, in each case 40 mg/l
Rosa sp. (roses) Geraniol Nerol, citronellol [157]
Solanum aviculare Citronellal, 150 mg/l Menthane-3,8-diol, citronellol, isopulegol, 15–39% transformation [160]
Limonene, 1.2 g/l Carvone (240 mg/l), via carveol [141]
Spirodela punctata Alkyl substituted citronellol Hydroxyl derivatives [161]
Triticum aestivum, seeds Neral, geranial, citronellal, pulegone, Reduced to the corresponding alcohols and oxidized to the corresponding acids, total up to 21 mM in the seeds [162]
carvacrol, vanillin, 40 ml/ml in the vial air
Vanilla planifolia Ferulic acid Vanillin [163]
1 mM 3,4-Dihydroxy-benzaldehyde 16.7 mg vanillin and 286 mg vanillyl alcohol per 100 g DW embryo culture [164]
Fungi and algae
Aspergillus niger b-Ionone Hydroxyl and oxo derivatives [165]
(–)-Guaiol The aroma material pancherione [166]

View this article online at wileyonlinelibrary.com

Nerol, geraniol, 0.1% v/v Linalool, 43 mg/l, a-terpineol 54 mg/l [167]
Capric acid, caprilic acid Up to 46% conversion to nonan-2-one and heptan-2-one [168]
Phenylalanine 2-Phenylethanol [169]
Aspergillus niger followed by Ferulic acid Vanillin, 767 mg/l [170]
Pycnoporus cinnabarinus
Beauveria bassiana Cadina-dien-3-one Various hydroxylated derivatives [171]
Botryodiplodia theobromae a-Linolenic acid Jasmonic acid, 1100 mg/l (100 mg/g dry cells) [82]
[111]
Botryodiplodia theobromae (Mucor fungus) (+)-Valencene (sesquiterpene), 0.4 g/l Nootkatone [172]
Botryosphaeria dothidea (Mucor fungus) (+)-Valencene (sesquiterpene), 0.4 g/l Nootkatone [172]
Candida boidinii Betuligenol Raspberry ketone [173]
Candida sorbophila Ricinoleic acid g-Decalactone, 40.9 g/l [174]
Chaetomium globosum Valencene Nootkatone, 25 mg/l [175]
Chlorella sp. (algae) (+)-Valencene (sesquiterpene), 0.4 g/l Nootkatone, 90% conversion [172]
Cladosporium suaveolens 5% Ricinoleic acid g-Decanolide, 0.5% [176]
Curvularia lunata Squamulosone (aromadendranone), 0.16% w/v Various aromadendranes, <0.06% w/v [177]
Dunaliella tertiolecta (halophilic green algae) Terpene aldehydes Reduction of the aldehydes to alcohols [178]
Epicocum nigrum Citrus sinensis oil Terpene alcohols [179]

Copyright © 2010 John Wiley & Sons, Ltd.

Fusarium proliferatum Limonene, 0.5% Carveol, 35 mg/l [180]
Fusarium verticilloides Limonene Perillyl alcohol [181]
Ganoderma applanatum Myrcene, 0.1% v/v Myrcenol and other volatile oxygenated metabolites [182]
Glomerela singulate b-Selinene Hydroxylation at three positions [183]
Gomgronella butlery Limonene, 0.5% Perillyl alcohol, 16 mg/l [180]
Ischnoderma benzoinum b,b-Carotene, 20 mg/l Flavour compounds (b-ionone, b-cyclocitral, dihydroactinidiolide, hydroxyl-trimethyl-cyclohexanone), total [184]
<6 mg/l
Kluyveromyces marxianus Phenylalanine 2-Phenylethanol, 26.5 g/l, 2-phenylethylacetate, 6.1 g/l [185]
Marasmius scorodonius b,b-Carotene, 20 mg/l Flavour compounds (b-ionone, b-cyclocitral, dihydroactinidiolide, hydroxyl-trimethyl-cyclohexanone), total [184]
<6 mg/l
Mucor circillenoides Ethyl decanoate, ethyl caprinate g-Decalactone, 10.5 g/l [186,187]
Penicillium citrinum Limonene, 0.5% a-Terpineol, 80 mg/l [180]
Penicillium digitatum Limonene, 0.5% a-Terpineol, 506 mg/l [180]
Penicillium digitatum, spores Geraniol, nerol, citral, 1 mM Methylheptenone, 0.9 mM [188]
Penicillium roqueforti, spores Octanoic acid, 0.5 mM 2-Heptanone, 0.48 mM [189]
Phanerochaete chrysosporium Phenylalanine, cinnamate Benzaldehyde [190]
Pichia pastoris 2–5% Alcohols (propanol, butanol, hexanol, etc.) 20–50% conversion to the corresponding flavour aldehydes (propionaldehyde, butyraldehyde, hexanal, etc.) [191]
Pleurotus flabellatus Myrcene, 0.1% v/v Myrcenol and other volatile oxygenated metabolites [182]
Pleurotus sajor-caju Myrcene, 0.1% v/v Myrcenol and other volatile oxygenated metabolites [182]
Y. Gounaris

Flavour Fragr. J. 2010, 25, 367–386

 Table 4. Continued

Organism Substrate Product/effect/yield Reference

Pleurotus sapidus Limonene, 0.5% Carvone [180]
Pleurotus sapidus Limonene, 0.5% v/v Carveol (30 mg/l), carvone (55 mg/l) [192]
Polyporus tuberaster Phenylalanine, 45 mM Benzaldehyde, 11.93 mM [193]
Saccharomyces cerevisiae Aliphatic aldehydes, 2-keto-carboxylic acids Acyloins [194]
Thiol precursors Three times more flavour thiols in wine fermentation [195]
Massoi oil d-Decalactone, 7.45 g/l [196]
Phenylalanine 2-Phenylethanol, 7 g/l [197]

Flavour Fragr. J. 2010, 25, 367–386

Oleic acid 2-Phenylethanol, 12.6 g/l [198]
Terpenoid glycosides Free terpenoids [199]
Sporidiobolus salmonicolor Ricinoleic acid, 5 g/l g-Decalactone, 40 mg/l [107]
Ricinoleic acid g-Decalactone, 131.8 mg/l [200]
Sporidiolobus ruinenii Methyl recinolate g-Decalactone, 5.5 g/l [201]
Torulopsis bombicola Palmitic acid Hexadecanolide lactone (musk fragrance) [202]
Trametes versicolor b,b-Carotene, 20 mg/l Flavour compounds (b-ionone, b- cyclocitral, dihydroactinidiolide, hydroxyl-trimethyl-cyclohexanone), [184]
total <6 mg/l
Trametes suaveolens Phenylalanine Benzaldehyde [203]
Yarrowia lipolytica Methyl ricinoleic acid 5 g/l g-Decalactone, g-octalactone, g-dodecalactone [204,205]
Zygorhynchus moelleri 0.2% Vanillic acid Vanillyl alcohol, 5 g/l [94]
Bacteria
Alcaligenes defragrans Limonene + isolimonene (or phelandrene), 1 mM each Isoterpinolene, 250 mM [206]
Biotechnology for essential oils, flavours and volatile isolates

Amycolatopsis sp. Ferulic acid Vanillin, 12 g/l [207]

Clostridium tyrobutyricum 5-Hydroxy-2-decenoic acid or 5-hydroxy-2-dodecenoic acid 5.95 g/l d-Decalactone or 13 g/l d-dodecalactone [208]
Bacillus sp. Lutein, 0.04 mM Ionones, total <10 mg/l [209]
Bacillus fusiformis 50 g/l Isoeugenol Vanillin, 8.10 g/l [210]
Calothrix (Cyanobacteria) Norcarotenoids, 0.1–0.4 mM Pentan- and hexan-diones, hydroxypentanones and hydroxyhexanones [211]
Bacillus subtilis 1% Massoia lactone 40% Transformation to d-decalactone [212]
Geotrichium sp. Lutein, 0.04 mM Ionones, total <10 mg/l [209]
Ischnoderma benzoicum Phenylalanine Benzaldehyde, 0.3 g/l [213]
Plectonema (Cyanobacteria) Norcarotenoids, 0.1–0.4 mM Pentan- and hexan-diones, hydroxypentanones and hydroxyhexanones [211]
Rhodococcus sp. Geraniol Geranic acid [214]
Xanthobacter sp. C20 Limonene, 1 mM Limonene-8,9-epoxide, 0.8 g/l [215]

Copyright © 2010 John Wiley & Sons, Ltd.

Pseudomonas fluorescens a-Pinene oxide, 0.01–0.03 g/l Isonovalal, 108 g/l culture. [216]
Pseudomonas putida Ferulic acid, Vanillic acid [217]
Ferulic acid + dithiotheitol Vanillin, 210 mg/l [218]
Isoeugenol, 30 mM Vanillin, 10 mM [219]
Pseudomonas rhodesiae a-Pinene oxide Isonovalal, 16.4 g/l/g biomass, 160 g/l/h [220]
Pseudomonas sp. Pinenes, 1% v/v Cymene (130 mg/l), terpineol, borneol, limonene, terpinolene (all <40 mg/l) [221]
2% Massoia lactone Up to 85% conversion to d-decalactone [212]
Pycnoporus cinnabarinus Vanillic acid Vanillin, 2.5 g/l [222]
Rhodococcus erythropolis Carveol, 50 mM Carvone, 160 mM [223]
Rhodococcus opacus Limonene Carveol, carvone [224]
[225]
Serratia marcescens 0.4% Eugenol Vanillin, 3.8 g/l [226]
Streptomyces setonii Ferulic acid, vanillic acid Vanillin, 64 g/l [227]
Ferulic acid Vanillin, 13.9 g/l [228]
Streptomyces sp. V-1 45 g/l Ferulic acid Vanillin, 19.2 g/l [229]

The main products are shown. Yields are cited for the optimum culture time and conditions if reported explicitly, as in many cases the products are referred as simply ‘detected’.

View this article online at wileyonlinelibrary.com

375

soluble additives. b-Cyclodextrins are water-soluble molecular
cages that, properly modified, could sequester the desired
product into their cavities. In addition, this confinement of the
product into the clathrate cage protects it from exposure to
degrading conditions in the culture medium.
Of the volatiles listed in Table 4, vanillin is of the highest
importance, followed by benzaldehyde, capsaicin, g-decalactone
and 2-phenylethanol. The commercial aspects, the organoleptic
and bioactivity qualities, the uses and the production method-
ologies of these volatiles have been discussed by Cheetham,[202]
Dignum et al.,[1] Garavaglia et al.,[91] Krings and Berger,[118]
Ramachandra and Ravishankar,[41] Schade et al.[231] and
Vandamme and Soetaert.[78] Vanillin is produced at about 12 000
tonnes annually, primarily from guaiacol. Only 20 tonnes are
produced by extraction from vanilla beans. It is used as a food
additive, but also possesses antimicrobial, antioxidant and anti-
mutagenic qualities. The biotransformation of ferulic acid to van-
illin using Amycolatopsis can produce up to 12 g/l culture
medium,[207] whereas yields in the range 13–>60 g/l have been
achieved with Streptomyces species. [227–229] Capsaicin, the major
pungent compound in chilli peppers, is produced by the same
biosynthetic path as vanillin. It is used as a food additive and a
counter-irritant in rheumatisms and neuralgias. Of commercial
interest could be the method of producing it using immobilized
placenta cultures of Capsicum frutescens, biotransforming cou-
maric acid.[232] The flavour compound benzaldehyde is the
second most important volatile. Currently, it is obtained form
amygdalin. Biotransformation of phenylalanine to benzaldehyde
by Ischnoderma benzoinum seems to be a possible alternative,
since yields of 1 g/l can be achieved by in situ recovery meth-
ods.[118,213] The peach-smelling g-decalactone, product of lipid
catabolism, is also easily made using chemical synthesis. The
biotransformation of ricinoleic acid by Candida sorbophila can
produce up to 40 g/l g-decalactone;[174] Mucor circillenoides
achieves over 10 g/l from fatty acid esters;[186,187] and Yarrowia
lipolytica produces 5 g/l of a mixure of g-lactones, mainly
g-decalactone, from methyl ricinoleic acid.[204,205] These values are
much higher than the theoretical limit of about 1 g/l required for
consideration of a procedure as an alternative commercially
interesting process. The same holds for the ‘rose’ flavour com-
pound 2-phenylethanol, product of the phenylpropanoid path,
obtained by biotransformation of phenylalanine. Kluyveromyces
marxianus, cultured in the presence of phenylalanine, produces
over 26 g/l 2-phenylethanol[123,185] and Sacharomyces cerevisiae
yields 12.6 g/l from oleic acid and 7 g/l from phenylalanine.[198]
An inspection of Tables 3 and 4 shows the superiority of micro-
orgamisms compared to cultured plant cells, as far as yields of
volatile compound production are concerned.
Biotransformations by Isolated Enzymes
Isolated enzyme systems often have a higher biotransformation
rate than cell-based procedures, but the cost of isolating the
enzyme is a basic factor determining their employment.[233]
Enzymes exhibit high stereo-selectivity in the conversion
process, minimizing the production of undesirable isomer and
enantiomer by-products often accompanying cell-mediated
biotransformations.[234] However, only about 7% of the studies
on biotransformation involve isolated enzymes, the rest being
concerned with whole-cell systems.[125] Although they have
been applied to a significant extent,[126] only a few cases of
376
biotransformations involve volatiles. Some examples are given
View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd.
Y. Gounaris
in Table 5. The yields are improved by immobilizing the enzyme
on polymer, glass or other supports, or by encapsulating it inside
membranes. The resulting easy recoverability and re-use of the
enzyme lowers the cost of operation. Overall, and with the
exceptions of glycosidases and esterases, most of the enzymes
used in bioconversions catalyse the introduction of some
oxygen function in the substrate molecule. This reduces its vola-
tility, unless it splits the molecule into smaller volatile parts.
The majority of the isolated enzyme-catalysed bioconversions
involve lipid-derived volatile products. In the phenolic and ter-
penoid category, the most promising use is in liberating volatiles
from their glycosylated forms. Of particular importance, as far as
yields are concerned, is the formation of ‘green’ note flavours,
hexenols and hexenals, from fatty acids by the action of hetero-
geneous crude enzyme preparations[262] and the liberation of
vanillin from glycovanillin by the action of commercial pectinase
and cellulase.
Metabolic Engineering
Genetic engineering of metabolic paths is the newest and most
promising effort for improvement of volatile production. There
are several excellent reviews on metabolic engineering of the
secondary metabolite pathways in general[263–266] and also on
terpenoid paths specifically.[267–269] This review concentrates
on cases concerning the production of volatile compounds.
Genetically Engineered Microorganisms
Bacteria and yeasts have been genetically altered, either to
improve their synthesis of volatiles or to produce new ones the
microorganism is normally unable to synthesize. A list of promis-
ing examples is given on Table 6. Microorganisms are particularly
well suited for fermentation and general culturing procedures.
Genetically manipulating them to produce volatiles from raw
substrates, in commercially worthwhile quantities, would be a
major breakthrough in biotechnology. Although most of the
achieved yields are still in the mg/l scale, vanillin can be produced
at 5 g/l by E. coli carrying the 3-deoxyarabino-heptulosonic acid
7-phosphate synthase gene[271] and amorphadiene yields of
37 g/l have been observed with S. cerevisiae transformed with
operons of the methyl–erythritol and mevalonate path genes.[283]
Yarrowia lipolytica with genetically engineered genes of the
b-oxidation genes is able to reach 10 g/l g-decalactone produc-
tion from ricinoleic acid esters.[285–289] Volatile dicarboxylic acids
can accumulate at amounts up to 80 g/l, when various yeasts
having genetically interrupted b-oxidation and amplified
w-hydroxylation were fed with fatty acids and alkanes.
Genetically Engineered Plants
Terpenoids. A compilation of some of the metabolic engi-
neering efforts concerning the monoterpene- and
sesquiterpene-synthesizing paths is presented in Table 7. The
CaMV35S promoter was commonly used in such studies.
However, expression of the transgene often proved tissue-
specific. Co-suppression instead of overexpression was also a
common occurrence. Most successful was the overexpression of
the DXR of the MEP path, where a 50% increase in total essential
oil yield in peppermint was attained.[293] Monoterpene cyclases
are slow enzymes and higher yields of cyclic monoterpenes
were achieved by overexpressed limonene synthase, especially
Flavour Fragr. J. 2010, 25, 367–386
 Table 5. Biotransformations involving volatile compounds, effected by crude or purified enzyme preparations

Enzyme/extract Source Product/results Reference

100 kDa cut-off extract Tomato fruits 5 mg/min hexanal from 16 mM linoleic acid [235]
Alcohol dehydrogenase Yeast Production of 1-phenyl-2-propanol from 1-phenyl-2-propanone [236]
Lactobacillus kefir Synthesis of phenylethanol from acetophenone [237]
Amine oxidase Aspergillus niger Production of vanillin from vanillamine [238]
Benzaldehyde lyase, immobilized Pseudomonas fluorescens Traces of benzoin produced from benzaldehyde [239]
Cell-free extract Potato tuber Hexenals, hexanal, pentenals, pentenol were formed from fatty acid [240]

Flavour Fragr. J. 2010, 25, 367–386

hydroperoxides
Cutinase, immobilized Fusarium solani Alcoholyses and hydrolyses esters of volatile alcohols [241]
Cyclase Cichorium intybus (chicory) Cyclized germanone-4,5-epoxide converted into neoprocureumenol [242]
Enzyme mix Citrus poonensis fruit peel 30% of limonene was converted to terpineol, linalool, and linalool [243]
oxide
Germacrene A hydroxylase Cichorium intybus Production of nootkatone from valencene [244]
Hydroperoxide lyase and alcohol Saccharomyces cerevisiae (whole cell extracts. Transformed with Production of cis-3-hexenal, cis-3-hexenol from lipid degradation [245]
dehydrogenase hydroperoxide lyase was from banana)
Hydroperoxide lyase Excreted to the medium by Pichia pastoris, transgenic with the 30 mg/l hexanal and nonenal from C18 hydroxy-fatty acids. [246]
tomato hydroperoxide lyase
Excreted to the medium by Yarrowia lipolytica, transgenic with the Accumulation of 350 mg/l C6 aldehydes in the culture medium, from [247]
green pepper hydroperoxide lyase. Lipid-induced promoter olive oil and fatty acids
Laccase Various microorganisms 28.6% transformation of valencene to nootkatone [248]
Biotechnology for essential oils, flavours and volatile isolates

Lipases Various microorganisms Hydrolyse menthyl esters, liberating menthol [234]

Lipoxygenase/hydroperoxide lyase
Lipoxygenase, hydroperoxide lyase
Membrane fraction, immobilized
Microsomes
Catalyse synthesis of flavour esters [249–251]
Apple pomace, crude mixture Production of hexanal and 2,4-decadienal from unsaturated fatty [252]
acids
soybean, recombinant hydroperoxide lyase Production of hexanal and hexenal from vegetable oils [253]
Soya Used in combination, converted polyunsaturated fatty acids into C6 [254]
aldehydes and alcohols
Carnation flowers, tomato leaves, strawberry leaves Production of hexanal from linoleic acid [231]
Nicotiana tobacco Hydroxylated terpineols at allylic positions. Use NADPH and O2. [153]
Probably cyt P-450-dependent monoxygenases

Copyright © 2010 John Wiley & Sons, Ltd.

Viscozyme (pectinase) + Celluclast Commercial enzymes Transformation of glucovanillin from vanilla pods into vanillin. 1.8 g [255]
(cellulase) vanillin from 100 g dry pods
Peroxidase Horseradish Introduction of hydroxyl and methoxy groups at the 6,7 double bond [256]
of citronellol
Phenoloxidase Mucuna pruriens Produced catechols from monophenols [257]
a-Acetolactate decarboxylase, Yeast 12-Day faster conversion of a-acetolactate and diacetyl into acetoin [258]
encapsulated in beer
b-Glycosidases Grapes, yeasts, bacteria, fungi Hydrolyse monoterpenoid glycosides of plant juices, liberating the [199]
monoterpenoids [259]
[260]
Almond Benzaldehyde production from amygdalin [202]
Immobilized sesquiterpene hydroxylases Chicory 50–100% Transformation of sesquiterpenes to the corresponding [261]
and sesquiterpene alcohol alcohols, aldehydes and acids
dehydrogenases
Mixture of materials (soya for Hydrolysed oils (mainly linoleic and linolenic acid) 4.2 g/kg 3(Z)-hexen-1-ol, 1.5 g/kg 2(E)-hexenal [262]
lipoxygenase, guava for
hydroperoxide lyase, baker’s yeast for
alcohol dehydrogenase)

View this article online at wileyonlinelibrary.com

377
 378

Table 6. Production of volatile matabolites by transgenic microorganisms

Microorganism Transgene/genetic manipulation Products Reference

E. coli With DXP synthase, IDP isomerase, FDP synthase, Pinene, myrcene, terpinene, carene, sabinene, limonene, [270]
monoterpene cyclase terpinolene
3-Deoxyarabino-heptulosonic acid 7-phosphate synthase Vanillic acid, 5 g/l [271]
and catechol-O-methyl-transferase
Mevalonate path basic gene operons [272]
Engineered mevalonate path and other terpenoid Produces IPP, DMAPP, casbene (if the GGPP synthase gene is also [273]
synthesis genes included), 2.2 mg/l amorphadiene (if the FPP synthase gene is
included). Mevalonate included in the substrate in some cases
Sesquiterpene synthase Up to 8.9 mg/l geranyl geraniol, when mutant GGPP synthase [274]
and mutant HMG-CoA synthase were used. Up to 22–26 mg/l

View this article online at wileyonlinelibrary.com

when mutant FPP synthase or IPP isomerase were used under
IPP or DMAPP feeding
Feruloyl-CoA synthetase and feruloyl Synthesis of patchoulol [275]
Hydratase/aldolase from Pseudomonas fluorescens 70.6% Conversion to vanillin (2.5 g/l). 3.3 mM ferulic acid as
substrate
A 9.6 kb DNA fragment of Bacillus stearothermophilus [276]
Bacillus stearothermophilus limonene methyl oxidase or 33 mg/l Terpineol, 26 mg/l carveol, 2.2 mg/l carvone from [277]
limonene hydratase limonene. 230 mg/l perillic acid and 36 mg/l perillyl aldehyde
from terpineol
29 mg/l Terpineol and 72 mg/l perillyl alcohol from the limonene
methyl oxidase transformants. 235 mg/l Terpineol and 35 mg/l
carvine with the hydratase transformants. Limonene included
in the substrate
Methylomonas sp. Limonene synthase Limonene, 0.5 mg/l [278]
Saccharomyces cerevisiae Clove DNA Eugenol, thymol, 2-phenylethyl alcohol [279]
epi-Cedrol synthase (eCS)or doubly by hmgr and mutated epi-Cedrol. 90 mg/l by ecs, 390 mg/l by ups2-1/hmgr [280]

Copyright © 2010 John Wiley & Sons, Ltd.

Yarrowia lipolytica
Various yeasts (Candida, Saccharomyces,
Schizosachcaromyces, Pichia, Yarrowia,
Sporobolomyces)
transcription factor ups2-1 (ups2-1/hmgr)
HMG-CoA reductase, GAL1 promoter
GPP synthase
Amorphadiene synthase [ADS], HMG-CoA reductase
[HMGR], FPP synthase
Various operons of the mevalonate and DXP path genes
Pinene hydroxylase
Acyl-CoA oxidase genes pox1, pox3 disrupted
Genetically engineered
Deleted acyl-CoA oxidases, selective for short chain fatty
acids
Genetically interrupted b-oxidation and amplified
w-hydroxylase
1 mg/l Abietadiene, geranygeranyldiphosphate [281]
Up to 10.6 mg/l geranylgeraniol [273]
Up to 700 mg/ml amorphadiene with ADS, 11.2 mg/ml with ADS [282]
and HMGR co-transformation. 61 mg/ml when met-repressible
FPP synthase was also introduced. 73 mg/ml amorphadiene
when FPP synthase was overexpressed
Up tp 37 g/l amorpha-4,11,-diene [283]
Biotransformation of pinene to myrtenol [284]
g-Decalactone, 0.35 g/l from methyl ricinoleate [285]
9.5 g/l g-calactone from methyl ricinoleate. [286]
10-Fold increase in g-decalactone production from fatty acids [287,288,289]
Up to 80 g/l flavour dicarboxylic acid from fatty acids or alkanes [290–292]
Y. Gounaris

Flavour Fragr. J. 2010, 25, 367–386

 Table 7. Production of volatile terpenoids by genetically engineered plants
Gene
Monoterpene path
1-Deoxy-xylulose-5-phosphate
reductoisomerase (DXR)
Limonene synthase (LS)
Flavour Fragr. J. 2010, 25, 367–386
Limonene-3-hydroxylase
(Lim3h)
Linalool synthase (LIS)
Linalool/nerolidol synthase
Copyright © 2010 John Wiley & Sons, Ltd.
Menthofuran synthase (MES)
Sesquiterpene path
germacrene A synthase
Nerolidol synthase
Sesquiterpene synthase TPS10
Trichodiene synthase
View this article online at wileyonlinelibrary.com
b-Farnesene synthase
379
Transgenic
Mentha piperita (peppermint)
Mentha piperita
Mentha arvensis and
Mentha piperita
Nicotiana tabacum (tobacco)
Mentha piperita
Nicotiana tabacum
Dianthus caryophyllus (carnation)
Lycopersicon esculentum (tomato)
Petunia hybrid
Arabidopsis thaliana
Petunia, potato, tomato,
Arabidospis
Mentha piperita
Arabidopsis thaliana
Arabidopsis thaliana
Solanum tuberosum (potato)
Arabidopsis thaliana
Nicotiana tabacum
Arabidopsis thaliana
Results/remarks References
Overexpression under CaMV 35S promoter gave 50% more essential oil. Same [293]
composition as in wild-type. Co-suppression of DXR results in less essential oil
Expression under CaMV 35S promoter. Transgenics had more menthone, menthofuran and [294]
pulegone and lower menthol than wild type plants. Only 1% increase in limonene
Under CaMV35S, no expression in glandular trichomes and no effect on oil yield and [295]
composition
Higher or lower levels of total monoterpenes, depending on the overexpression or [296]
co-suppression of the transgene correspondingly
Both, plastid-directed and cytosol-directed LS produced limonene reaching the [297]
concentrations of 143 ng/g fresh weight in the plastids and 40 ng/g in the cytosol
Under CaMV35S. Co-suppression. Percentage of limonene in the essential oil rose from 2% [295]
to 80%. No change in total oil yield
The plants already had a monoterpene synthase transgene. Hydroxylated limonene into [298]
isopiperitenol. From that, methatrienes, p-cymene and isopiperitenone were
synthesized
Expression under CaMV35S promoter. Linalool was produced in the flowers and leaves. [299]
Biotechnology for essential oils, flavours and volatile isolates
Petals had linalool oxide but not linalool. These monoterpenes are not found normally
in this plant
Under E8 late-ripening specific promoter. Synthesis of linalool and 8-hydroxylinalool in [300]
fruits
Formation of linalool, as glycoside, in leaves, sepals, petals, ovary, stem. Not detected in [301]
roots, pollen style and nectaries
Targeted to plastids. Synthesis of linalool and its glycosides. The glycosides were at levels [302]
up to 60-fold higher than those of free linalool. Also, synthesis of low levels of nerolidol
Up to 150 mg/g FW tissue linalool, its hydroxylated products and their glycosides [303]
Antisense silencing. 50% less menthofuran [293]
Overexpression. More menthofuran [304]
Expressed in the cytosol. Some germacrene A was produced. Low levels [302]
The transgene was introduced into the mitochondria. High levels of the sesquiterpene [305]
nerolidol were produced
The transgene was introduced into the plastids. The sesquiterpene nerolidol was [302]
synthesized
The transgene was introduced into the plastids. High levels of linalool, 8-hydroxylinalool [268]
and their glycosides were observed
Overexpression. Production of high quantities of farnesene bergamotene and other [306]
sesquiterpenes
Expressed under the CaMV 35S promoter. Low levels (5–10 ng/g FW) of trichodiene were [307]
detected
Synthesis of (E)-b-farnesene [308]
 Y. Gounaris

when directed to plastids. Although overexpression of rate-

References
limiting steps (DXR, cyclases) increased the total or specific yield

[313]
[309]
[310]

[311]

[312]

[314]
of monoterpenes, altering the expression of enzymes for non-
rate-limiting steps, or for more downstream steps, usually
altered their relative percentages. This might be advantageous if
the yield of the desired compound increases. Boosting the pro-
duction of monoterpene alcohols, as by overexpressed linalool

Suppression by RNA interference leads to strongly reduced levels (75–99%) of

Under CaMV35S promoter. The sense orientation was overexpressed 100-fold
synthase, also increases the rate of their glycosidation, so that

10-fold more of the volatiles 2-phenylacetaldehyde and 2-phenylethanol

Two AADCs decarboxylating phe and tyr were used. Transgenics produced
the yield of the glycosidated forms can far exceed that of the
free form. Overexpressing transgene prenyltransferases of ses-

Suppression by RNA interference leads to strongly reduced levels of

quiterpene synthesis, such as the farnesene, nerolidol and TSP10

and resulted in increased content of shikimate-derived volatiles

Under CaMV35S promoter. More benzyl and phenylethyl acetates
synthases, increases the linear and some cyclic sesquiterpenes
appreciably, and these can even be detected in plants not

Antisense suppression. 10–100-fold more methyl benzoate

normally producing them. More limited was the success with
non-regulated sesquiterpene cyclases, where only small
increases in the specific sesquiterpene yield were observed.

Shikimic acid-derived volatiles. Effects of both, overexpres-

sion and silencing of enzymes for synthesis of shikimate-derived
phenolic in transgenic plants are known (Table 8). Manipulating

shikimate-derived volatile benzenoids

the pre-tyrosine and pre-phenylalanine part of the shikimic acid
path does not seem very promising, due to the presence of mul-
tiple feedback-inhibited regulatory steps, which would nullify the
overexpression results. The same seems to be true for the pro-
duction of benzenoids from phenylpropanoids, although paths
not involving b-oxidation have been reported, whose regulation
characteristics, however, are still not defined.[315] At downstream

methylbenzoate
sites, overexpressions of aromatic amino acid decarboxylases, Table 8. Production of shikimic acid-derived volatile phenolics by genetically engineered plants

Results/remarks
alcohol dehydrogenase and alcohol acetyltransferase all resulted
in higher volatile phenolic yield. Reports on the effect of overex-
pression of transcription factor genes on volatile yield, as
reported by Memelink et al.[263] for flavonoid and indole alkaloids,
are not yet known for the shikimic acid path. This approach
seems, however, to be most promising and RNAi silencing of a
myb-type trans-factor in Petunia strongly reduced the levels of
volatile benzenoids.
Lycopersicon esculentum (tomato)

Lipid and amino acid-derived volatiles. Metabolic engineer- Dianthus caryophyllus (carnation)
ing of volatile production achieved its most spectacular results
Vitis vinifera (grapevine)

with the levels of aldehydes, alcohols and organic acid esters.

Examples are given in Table 9. For compounds produced by deg-
Transgenic plant

radation of unsaturated fatty acids, the largest increases (even

Petunia hybrida

Petunia hybrida

Petunia hybrida

five-fold) were observed by overexpression of desaturases and

lipoxygenases. Only moderate increases were achieved by over-
expressed hydroperoxide lyase. With alcohol dehydrogenase the
results were not unambiguous. In some cases, its upregulation
only increased the amount of alcohols, leaving the aldehydes
unchanged, and in other cases no significant changes were
observed for either type of compound. Since ethylene is involved
Aromatic amino acid decarboxylases

in the signal transduction path for stimulation of lipid-derived

aldehyde and alcohol synthesis, attempts were made to change
Alcohol acetyltransferase (AAT)
Alcohol dehydrogenase (ADH)

myb-Type trans factor (ODO1)

Benzoic/salicylic acid carboxy

the activity of 1-amino- cyclopropane-1-carboxylate synthase

methyltransferase (BSMT)

(ACS), the enzyme catalysing the synthesis of the ethylene pre-

Flavanone-3-hydroxylase

cursor 1-amino-cyclopropane-1-carboxylic acid. Up to now, only

downregulation of ACS has been reported, accompanied, as
expected, by drastic decreases in the levels of volatile aldehydes
and alcohols.
(AADC)

Acknowledgements
Gene

The author would like to thank his colleagues for their recom-
380

mendations on improving this article.

View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd. Flavour Fragr. J. 2010, 25, 367–386
 Table 9. Production of volatiles derived from catabolism of fatty acids or amino acids in transgenic plant species
Gene
1-Amino-cyclopropane-1-carboxylate
synthase (ACS)
1-Amino-cyclopropane-1-carboxylate
Flavour Fragr. J. 2010, 25, 367–386
oxidoreductase (ACC oxidase, ACO)
Acyl-CoA desaturases
Alcohol dehydrogenase (ADH)
Hydroperoxide lyase (HPL)
Copyright © 2010 John Wiley & Sons, Ltd.
Lipoxygenase (LOX)
2-Phenylethanol dehydrogenase
Ripening-associated membrane
protein (TRAMP)
View this article online at wileyonlinelibrary.com
381
Plant
Lycopersicon esculentum
(tomato)
Malus domestica (apple tree)
Cucumis melo var,
cantaloupensis cv.
Vedrantrais
(cantaloupe melon)
Malus domestica
Lycopersicon esculentum
Nicotiana tabacum
Lycopersicon esculentum
Arabidopsis thaliana
Lycopersicon esculentum
Solanum tuberosum
Lycopersicon esculentum
Nicotiana tabacum
Solanum tuberosum
Petunia hybrida
Lycopersicon esculentum
Results/remarks References
Downregulation of the synthase by its antisense form. Strong decrease in hexanal, [316]
hexanol, 3-hexanol, trans-2-heptenal in the fruits. No significant change in
cis-3-hexenal or trans-2-hexenal
Downregulation of the synthase by its antisense form. Strong decrease in volatile [317–319]
esters but not in volatile aldehydes
Downregulation of the oxidase by its antisense form. 60–85% reduction in total [320]
volatiles. Potent odorants (ethyl butanoate, ethyl 2-methylpropanoate, ethyl
2-methyl butanoate) were reduced by over 95%. Weak odorants (2-methylpropyl
acetate, 2-methyl butyl acetate) reduced by 50%
Downregulation of the oxidase by its antisense form. Production of butyl and hexyl [321]
acetate was blocked at the step of fatty acid reduction to aldehydes and at the step
of ester formation
Downregulation of the synthase by its antisense form. Strong decrease in volatile [317,319]
esters but not in volatile aldehydes
A yeast D9 desaturase was used. >3-fold increases were observed for [322]
1-hydroxy-2-butanone, 1-penten-3-ol, heptanal, 3-hexen-1-ol, 2-octanol,
cis-3-hexenal, hexanal, 2-nonenal. Also, increases in non-fatty acid-derived volatiles
Biotechnology for essential oils, flavours and volatile isolates
A yeast D9 and an insect D11 desaturases were used. More cis-3-hexenal, palmitoleic and [323]
palmitvaccenic acid
Overexpression of ADH2 under CaMV 35S or fruit-specific polygalacturonase promoter [324]
resulted in higher hexanol and (Z)-3-hexenol. Aldehydes remained unchanged.
Co-suppression results in lower amounts of these compounds.
No changes in volatile alcohols and aldehydes upon upregulation of the tomato ADH2. [325]
Downregulation resulted in less (Z)-3-hexenol but no change in (Z)-hexenal
The rice HPLs were targeted to Arabidopsis plastids. The transgenics produced low [326]
levels of aldehyde volatiles, but only in leaves and not in roots
A cucumber HPL, acting on 9-hydroperoxides, was overexpressed in tomatoes. Only [327]
small change in C9 aldehydes and alcohols
Antisense depletion of the HPL leads to greatly reduced levels of 3-hexenal and [328]
hexanal
LOX C of tomato was targeted to plastids. Under expression to 1–5% of normal, either [329]
by co-suppression of by antisense forms. Hexenal, hexenol reduced to 1.5% of
wild-type
LOX2 of soybean was overexpressed in tobacco under CaMV35S promoter. There was a [330]
50–529% increase in C6 volatile aldehyde production
The potato plastidic LOX H1 was silenced by antisense methodology. Volatile C6 [331]
aldehydes were reduced. No change in jasmonates
Five times more 2-phenylethanol [332]
TRAMP is a channel protein for water and small solutes. Overexpression suppressed [333]
production of hexenal, hexanal, hexenol, methylbutanol and
6-methyl-5-hepten-2-one. Antisense plants had more total volatiles

References
1. M. J. Dignum, W. J. Kerler, R. Verpoorte. Food Rev. Intl 2001, 17, 199.
2. F. Rohdich, J. Wungsintaweekul, W. Eisenreich, G. Richter, C. A.
Schuhr, S. Hecht, M. H. Zenk, A. Bacher. Proc. Natl Acad. Sci. USA
2000, 97, 6451.
3. J. M. Estévez, A. Cantero, A. Reindl, S. Reichler, P. León. J. Biol. Chem.
2001, 276, 22901.
4. F. Rohdich, W. Eisenreich, J. Wungsintaweekul, S. Hecht, C. A. Schuhr,
A. Bacher. Eur. J. Biochem. 2001, 268, 3190.
5. F. Rohdich, F. Zepeck, P. Adam, S. Hecht, J. Laupitz, T. Gräwert, S.
Amslinger, W. Eisenreich, D. Arigoni. Proc. Natl Acad. Sci. USA 2003,
100, 1586.
6. P. J. Proteau. Bioorg. Chem. 2004, 32, 483.
7. M. Wolfertz, T. D. Sharkey, W. Boland, F. Kühnemann. Plant Physiol.
2004, 135, 1939.
8. S. Gao, J. Lin, X. Liu, Z. Deng, Y. Li, X. Sun, K. Tang. J. Biochem. Mol.
Biol. 2006, 39, 502.
9. M. K. Julsing, M. Rijpkema, H. J. Woerdenbang, W. J. Quax, O. Kayser.
Appl. Genet. Mol. Biotech. 2007, 75, 1377.
10. I. Feussner, C. Wasternack. Ann. Rev. Plant Biol. 2002, 53, 275.
11. J. Sánchez, H. L. Harwood. Eur. J. Lipid Sci. Tech. 2002, 104, 564.
12. E. Combet, J. Henderson, D. C. Eastwood, K. S. Burton. Mycoscience
2006, 47, 317.
13. B. Höschle, V. Gnau, D. Jendroseek. Microbiology 2005, 151, 3649.
14. J. A. Aguilar, A. N. Zavala, C. Díaz-Pérez, C. Cervantes, A. L. Díaz-Pérez,
J. Campos-García. Appl. Environ. Microbiol. 2006, 72, 2070.
15. K. Förster-Fromme, B. Höschle, C. Mack, M. Bott, W. Armbruster, D.
Jendrossek. Appl. Environ. Microbiol. 2006, 72, 4810.
16. A. H. Scragg. Plant Cell Tiss. Org. 1995, 43, 163.
17. T. H. Kim, J. H. Shin, H. H. Baek, H. J. Lee. J. Sci. Food Agric. 2001, 81,
569.
18. C. M. Cotton, J. W. Gramshaw, L. V. Evans. J. Exp. Bot. 1991, 42, 377.
19. G. Reil, R. G. Berger. Z. Naturforsch. 1996, 51, 657.
20. R. P. Niedz, M. G. Moshonas, B. Peterson, J. P. Shapiro, P. E. Shaw. J.
Plant Cell Tiss. Org. 1997, 51, 181.
21. R. Agrawal, M. V. Patwardhan, K. N. Gurudutt. Biotech. Appl. Biochem.
1991, 14, 265.
22. G. Reil, R. G. Berger. J. Plant Physiol. 1997, 150, 160.
23. A. Giamakis, O. Kretsi, I. Chinou, G. Spyropoulos. Phytochemistry
2001, 58, 351.
24. P. K. Gupta, U. J. Mehta, A. F. Mascarenhas. Plant Cell Rep. 1983, 2,
296.
25. A. A. Gbolade, G. B. Lochwood. J. Plant Physiol. 1992, 140, 28.
26. G. Binder, A. A. Abou-Mandour, F. C. Czygan. Angew. Bot. 1999, 73,
105.
27. I. S. Chung, Y. M. Kang, J. M. Oh, T. Kim, H. J. Lee, Y. A. Chae. Biotech.
Tech. 1994, 8, 789.
28. P. V. Purohit, P. Khanna. In Plant Cell Culture in Crop Improvement,
S. K. Sen, K. L. Giles (eds). Plenum: New York, 1983, 377.
29. M. Williams, J. L. Harwood. Biochem. Soc. Trans. 1998, 26, S154.
30. M. Sokmen, J. Serkedijeva, D. Daferera, M. Gulluce, M. Polissiou, B.
Tepe, H. A. Akpulat, F. Sahin, A. Sokmen. J. Agric. Food Chem. 2004,
52, 3309–3312.
31. I. M. S. Alves-Pereira, M. Fernandes-Ferreira. Phytochemistry 1998, 48,
795.
32. G. Suvarnalatha, M. S. Naravan, G. A. Ravishankar, L. V. Venkatara-
man. J. Sci. Food Agric. 1994, 66, 439.
33. M. G. Lopez, I. R. Sanchez-Mendoza, N. Ochoa-Alejo. J. Agric. Food
Chem. 1999, 47, 3292.
34. P. S. C. Braga, P. C. Santos-Gomes, P. C. R. Valentao, P. B. Andrade, R. M.
Seabra, M. Fernandes-Ferreira. In Floriculture, Ornamental and Plant
Biotechnology, J. A. Teixeria da Silva (ed.). Global Science Books:
Isleworth, UK, 2006; 491.
35. B. Tirillini, B. Tosi. J. Essent. Oil Res. 1992, 4, 431.
36. Y. C. Hong, L. C. Huang, G. A. Reineccius, S. K. Harlander, T. P. Labuza.
Plant Cell Tiss. Org. 1990, 21, 245.
37. I. Zabetakis, J. W. Gramshaw. Food Chem. 1998, 61, 351.
38. M. Pletsch, S. Piacente, C. Pizza, B. V. Charlwood. Phytochemistry
1993, 34, 161.
39. I. Hook, H. Sheridan, G. Wilson. Phytochemistry 1991, 30, 3977.
40. H. Dornenburg, D. Knorr. Food Biotechnol. 1996, 10, 75.
41. R. S. Ramachandra, G. A. Ravishankar. Biotechnol. Adv. 2002, 20,
101.
42. D. V. Banthorpe, S. E. Barrow. Phytochemistry 1983, 22, 2727.
382
View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd.
Y. Gounaris
43. D. V. Banthorpe, S. A. Branch, V. C. O. Njar, M. G. Osborne, D. G.
Watson. Phytochemistry 1986, 25, 629.
44. S. W. Zito, V. Srivastava, E. Adebayo-Olojo. Planta Med. 1991, 57, 425.
45. E. Soler, G. Feron, M. Clastre, R. Dargent, M. Gleizes, C. Ambid. Planta
1992, 187, 171.
46. U. Vogeli, J. Chappell. Plant Physiol. 1988, 88, 1291.
47. U. Vogeli, J. Chappell. Plant Physiol. 1990, 94, 1860.
48. U. Vogeli, J. W. Freeman, J. Chappell. Plant Physiol. 1990, 93, 182.
49. K. L. Falk, J. Gershenzon, R. Croteau. 1990, 93, 1559.
50. A. M. Anterola, J. H. Jeon, L. B. Davin, N. G. Lewis. J. Biol. Chem. 2002,
277, 18272.
51. V. Seidel, J. Windhovel, G. Eaton, A. W. Alfermann, R. R. J. Arroo, M.
Medarde, M. Petersen, J. G. Wooley. Planta 2002, 215, 1031.
52. A. Mohammad-Babar, N. Singh, A. M. Shohael, E-J Hahn, K-Y Paek.
Plant Sci. 2006, 171, 147.
53. R. Moller, R. D. Ball, A. R. Henderson, G. Modzel, J. Find. Plant Cell Tiss.
Org. 2006, 85, 161.
54. T. B. Karyagina, O. A. Gaevskaya, E. A. Gukasova, T. V. Timchenko,
D. I. Bairamashvili. J. Plant Physiol. 2007, 54, 267.
55. K. Matsui, Y. Kaji, T. Kajiwara, A. Hatanaka. Phytochemistry 1996, 41,
177.
56. M. L. Fauconnier, A. Mouttalib, B. Diallo, M. Jaziri. J. Plant Physiol.
2001, 158, 953.
57. S. R. Chou. Dissert. Abstr. Int. B Sci. Eng. 1992, 3369.
58. M. L. Fauconnier, M. Jaziri, M. Marlier, J. Roggemans, J. P. Wathelet,
G. Lognay, M. Severin, J. Homes, K. Shimomura. J. Plant Physiol. 1993,
141, 759.
59. A. C. Figueiredo, M. Salome, S. Pais, J. J. C. Scheffer. Plant Cell Tiss.
Org. 1995, 40, 113.
60. A. Mandujano-Chavez, M. A. Schoenbeck, L. F. Ralston, E. Lozoya-
Gloria, J. Chappell. Arch. Biochem. Biophys. 2000, 381, 285.
61. I. Saad, E. Diaz, I. Chavez, R. Reyes-Chilpa, A. Rubluo, M. Jimenez-
Estrada. Phytochemistry 2000, 55, 51.
62. S.-W. Shin, Y.-S. Kim, C.-A. Kang. Nat. Prod. Sci. 2001, 7, 120.
63. A. C. Figueiredo, J. G. Barroso, L. G. Pedro, J. J. C. Scheffer. In Floricul-
ture, Ornamental and Plant Biotechnology: Advances and Topical
Issues, vol. II, J. A. Teixeira da Silva (ed.). Global Science Books:
Isleworth, UK, 2006; 478.
64. M. Kino-Oka, H. Nagatome, M. Taya. Adv. Biochem. Eng. 2001, 72,
183.
65. M. I. Georgiev, A. I. Pavlov, T. Bley. Appl. Microbiol. Biotechnol. 2007,
74, 1175.
66. S. Srivastava, A. K. Srivastava. Crit. Rev. Biotechnol. 2007, 27, 29.
67. T. S. Lu, F. J. Parodi, D. Vargas, L. Quijano, E. R. Metrooetomo, M. A.
Hjortso, N. H. Fischer. Phytochemistry 1993, 33, 113.
68. P. A. G. Santos, A. C. Figueiredo, P. M. R. Lourenco, J. G. Barroso, L. G.
Pedro, M. M. Oliveira, J. Schripsema, S. G. Deans, J. J. C. Scheffer.
Biotechnol. Lett. 2002, 24, 1031.
69. A. I. Kennedy, S. G. Deans, P. K. Svoboda, A. I. Gray, P. G. Waterman.
Phytochemistry 1993, 32, 1449.
70. E. Szoke, E. Maday, S. A. Kiss, L. Sonnewend, E. Lemberkovics, M. S.
Seelig, R. Vink. Am. Coll. Nutr. 2004, 23, 763S.
71. O. Olszowska, A. W. Alfermann, M. Furmanowa. Plant Cell Tiss. Org.
1996, 45, 273.
72. Y. Matsuda, H. Toyoda, A. Sawabe, K. Maeda, N. Shimizu, N. Fujita,
T. Fujita, T. Nonomura, S. Ouchi. J. Agric. Food Chem. 2000, 48,
1417.
73. N. Comey, I. Hook, H. Sheridan, J. Walsh, P. James. J. Nat. Prod. 1997,
60, 148.
74. P. A. G. Santos, A. C. Figueiredo, M. M. Oliveira, J. G. Barroso, L. G.
Pedro, S. G. Deans, J. J. C. Scheffer. Plant Sci. 2005, 168, 1089.
75. P. M. Santos, A. C. Figueiredo, M. M. Oliveira, J. G. Barroso, L. G. Pedro,
S. G. Deans, A. K. M. Younus, J. J. C. Scheffer. Phytochemistry 1998,
48, 455.
76. F. Peraza-Luna, M. Rodriguez-Mendiola, C. Arias-Castro, J. M. Bes-
siere, G. Calva-Calva. J. Agric. Food Chem. 2001, 49, 6012.
77. F. Granicher, P. Christen, I. Kapetanidis. Phytochemistry 1995, 40,
1421.
78. E. J. Vandamme, W. Soetaert. J. Chem. Technol. Biotechnol. 2002, 77,
1323.
79. L. Zheng, P. Zheng, Z. Sun, Y. Bai, J. Wang, X. Guo. Biores. Technol.
2007, 98, 1115.
80. L. Janssens, H. L. de Pooter, E. J. Vandamme, N. M. Schamp. Process.
Biochem. 1992, 27, 195.
81. M. I. Farbood, B. J. Willis. US Patent No. 4560656, 1985.
Flavour Fragr. J. 2010, 25, 367–386
Biotechnology for essential oils, flavours and volatile isolates
82. F. Eng, M. Gutiérrez-Rojas, E. Favela-Torres. Process. Biochem. 1998,
33, 715.
83. A. B. P. Madeiros, A. Pandey, L. P. S. Vanderberghe, G. M. Pastore, C. R.
Soccol. Food Technol. Biotech. 2006, 44, 47.
84. B. Stefer, M. Schilling, B. Kunz. In Proceedings of the 17th Forum for
Applied Biotechnology, Universiteit Gent, Belgium, 2003; 247.
85. W. Bluemke, J. Schrader. Biomol. Eng. 2001, 37, 137.
86. A. L. Demain, M. Jackson, N. R. Trenner. J. Bacteriol. 1967, 94,
323.
87. S.-M. Wu, H. Zorn, U. Krings, R. G. Berger. Flavour Fragr. J. 2007,
22, 53.
88. N. Mdaini, M. Gargouri, M. Hammami, L. Monser, M. Hamdi. Appl.
Biochem. Biotech. 2006, 128, 227.
89. L. Janssens, H. L. Depooter, N. M. Schamp, E. J. Vandamme. Process.
Biochem. 1992, 27, 195.
90. F. Drawert, H. Barton. J. Agric. Food Chem. 1978, 26, 765.
91. J. Garavaglia, S. Hickmann Flôres, T. M. Pizzolato, M. do Carmo
Peralba, M. A. Záchia Ayub. World J. Microb. Biol. 2007, 23, 1273.
92. A. B. Medeiros, A. Pandey, R. J. Freitas, P. Christen, C. R. Soccol.
Biochem. Eng. J. 2000, 6, 33.
93. G. Laufenberg, P. Rosato, B. Kunz. Eur. J. Lipid Sci. Tech. 2004, 106, 207.
94. P. S. J. Cheetham, M. L. Gradley, J. T. Sime. US Patent No. 6844019 B1,
2005.
95. B. Gross, A. Gallois, H.-E. Spinnler, D. Langois. J. Biotech. 1989, 10,
303.
96. K. Okamoto, M. Chimori, F. Iwanaga, T. Hattori, H. Yanase. J. BioSci.
Bioeng. 2002, 94, 182.
97. T. Kawabe, H. Morita. J. Agric. Food Chem. 1993, 41, 637.
98. R. G. Berger, S. Hadrich-Meyer, F. Drawert. In Proceedings of the 11th
International Congress of Essential Oils, Fragrances and Flavours, vol.
3, S. C. Bhattacharyya, N. Sen, K. L. Sethi (eds). Aspect Publishing:
London, UK, 1990; 127.
99. P. Deetae, P. Bonnarme, H. E. Spinnler, S. Helinck. Appl. Microb. Cell
Physiol. 2007, 76, 1161.
100. P. Christen, A. Bramorski, S. Revah, C. R. Soccol. Biores. Technol. 2000,
71, 211.
101. A. Mohamed, J. Destain, M. Aguedo, L. Majad, H. Ghalfi, J-P. Wathelet,
P. Thonart. Appl. Biochem. Biotechnol. 2009, 158, 41.
102. S. Favale, P. Pietromarchi, G. Ciolfi. Vitis 2007, 46, 39.
103. C. Cerny, T. Huynh-Ba, W. Matthey-Doret. US Patent No. 5948453,
1999.
104. A. Tsakiris, V. Sipsas, A. Bekatorou, A. Mallouchos, A. A. Koutinas. J.
Agric. Food Chem. 2004, 52, 1357.
105. L. Del Toro-Sánchez, S. Sánchez, M. A. Ortiz, S. Villanueva, E. Lugo-
Cervantes. Appl. Microb. Cell Physiol. 2006, 72, 155.
106. M. E. Guerzoni, P. Vernocchi, M. Ndagijimana, A. Gianotti, R. Lanciotti.
Food Microbiol. 2007, 24, 139.
107. G. M. de Billerbeck, M. Menier, P. Raguenaud, C. Ambid. In Flavour
Research at the Dawn of the Twenty-first Century. Proceedings of
the 10th Weurman Flavour Research Symposium, J. L. Le Quere, P. X.
Etievant (eds). Editions Tec & Doc: Paris, France, 2003; 377.
108. S.-L. Lee, S. Juslin, C.-C. Chou. J. Ferment Bioeng. 1996, 82, 42.
109. C. V. de Vos Petersen, H. C. Beck, F. R. Lauritsen. Biotech. Bioeng. 2004,
85, 298.
110. J. S. Dickschat, H. B. Bode, S. C. Wenzel, R. Müller, S. Schulz. Chem.
Biochem. 2005, 6, 2023.
111. A. Häusler, T. Munch. Am. Soc. Microbiol. News 1997, 63, 551.
112. M. Etschmann, D. Sell, J. Schrader. In Flavour Research at the Dawn of
the Twenty-first Century. Proceedings of the 10th Weurman Flavour
Research Symposium, J. L. Le Quere, P. X. Etievant (eds). Editions Tec &
Doc: Paris, France, 2003; 385.
113. L. Serrano-Carreón, C. Flores, E. Galindo. Biotechnol. Prog. 1997, 13,
205.
114. A. A. de Araujo, J. M. Pastore, R. G. Berger. Appl. Biochem. Biotechnol.
2002, 98, 747.
115. G. F. Kapfer, R. G. Berger, F. Drawert. Biotechnol. Lett. 1989, 11, 561.
116. J. Rabenhorst, J. Gatfield J. US Patent No. 6451565, 2002.
117. L. Janssens, H. L. Depooter, L. De Mey, E. J. Vandamme, N. M.
Schamp. Mededelingen van de Faculteit Landbouwwetenschappen,
Universiteit Gent 1989, 54, 1387.
118. U. Krings, R. G. Berger. Appl. Microbiol. Biotechnol. 1998, 49, 1.
119. M. A. Longo, M. A. Sanromán. Food Technol. Biotechnol. 2006, 44,
335.
120. M. Gopipath, L. Vijayakumar, R. Dhanasekar, T. Viruthagiri, G. Baskar.
Global J. Biotech. Biochem. 2008, 3, 60.
Flavour Fragr. J. 2010, 25, 367–386 Copyright © 2010 John Wiley & Sons, Ltd.
121. S. ur Rehman, A. Paterson, J. R. Piggott. Trends Food Sci. Technol.
2006, 17, 557.
122. S. Serra, C. Fuganti, E. Brenna. Trends Biotechnol. 2005, 23, 173.
123. J. Schrader, M. M. W. Etschmann, D. Sell, J-M. Hilmer, J. Rabenhorst.
Biotechnol. Lett. 2004, 26, 463.
124. Y. Waché, F. Husson, G. Feron, J-M. Belin. A. Van Leeuw. J. Microb.
2006, 89, 405.
125. C. C. C. R. de Carvalho, M. M. R. da Fonseca. Biotechnol. Adv. 2006, 24,
134.
126. A. Giri, V. Dhingra, C. C. Giri, A. Singh, O. P. Ward, M. L. Narasu.
Biotechnol. Adv. 2001, 19, 175.
127. K. Ishihara, H. Hamada, T. Hirata, N. Nakajima. J. Mol. Catal. B 2003,
23, 145.
128. Z. M. Everitt, G. B. Lockwood. Fitoterapia 1992, 63, 534.
129. L. Yang, J. Dai, J. I. Sakai, M. Ando. J. Asian Nat. Prod. Res. 2006, 8,
317.
130. S.-M. Kang, H.-Y. Jung, Y.-M. Kang, J-Y. Min, C. S. Karigar, J-K. Yang,
S-W. Kim, Y-R. Ha, S-H. Lee, M-S. Choi. J. Agric. Food Chem. 2005, 53,
3449.
131. R. S. Ramachandra, G. A. Ravishankar. J. Biotech. 2000, 76, 137.
132. T. S. Johnson, G. A. Ravishankar, L. V. Venkataraman. Plant Cell Tiss.
Org. 1996, 44, 117.
133. H. Sakamaki, K.-I. Itoh, W. Chai, Y. Hayashida, S. Kitanaka, C. A.
Horiuchi. J. Mol. Catal. B 2005, 27, 177.
134. T. Hirata, Y. Ikeda, S. Izumi, K. Shimoda, H. Hamada, T. Kawamura.
Phytochemistry 1994, 37, 401.
135. H. Hamada, H. Yasumune, Y. Fuchikami, T. Hirata, I. Sattler, H. J. Wil-
liams, A. I. Scott. Phytochemistry 1997, 44, 615.
136. F. Carriere, G. Gil, P. Tapie, P. Chagvardieff. Phytochemistry 1989, 28,
1087.
137. T. Hirata, K. Koya, K. J. Sarfo, K. Shimoda, D. I. Ito, S. Izumi, S. Ohta, Y.
S. Lee. Mol. Catal. B 1999, 6, 67.
138. H. Hamada, Y. Fuchikami, Y. Ikematsu, T. Hirata, H. J. Williams, A. I.
Scott. Phytochemistry 1994, 37, 1037.
139. F. Drawert, R. G. Berger, R. Godelmann. Plant Cell Rep. 1984, 3, 37.
140. T. Kometani, H. Tanimoto, T. Nishimura, S. Okada. Biosci. Biotech.
Biochem. 1993, 57, 1290.
141. T. Vanek, I. Valterova, T. Vaisar. Phytochemistry 1999, 50, 1347.
142. Y. Orihara, T. Furuya. Phytochemistry 1993, 34, 1045.
143. K. Shimoda, Y. Kondo, T. Nishida, H. Hamada, N. Nakajima, H.
Hamada. Phytochemistry 2006, 67, 2256.
144. Y. Orihara, T. Fuyura, N. Hashimoto, Y. Degushi, K. Tokoro, T.
Kunisawa. Phytochemistry 1992, 31, 827.
145. Y. Orihara, K. Saiki, T. Furuya. Phytochemistry 1994, 31, 827.
146. Y. Orihara, T. Furuya. Phytochemistry 1994, 36, 55.
147. M. Shams-Ardakani, A. Ghannadi, P. Badr, A. Mohagheghzadeh.
Flavour Fragr. J. 2005, 20, 141.
148. S. Sakamoto, N. Tsuchiya, M. Kuroyanagi, A. Uno. Phytochemistry
1994, 35, 1215.
149. U. Werrman, D. Knorr. J. Agric. Food Chem. 1993, 41, 517.
150. S.-U. Kim, S.-H. Park. In Advances in Plant Glycosides: Chemistry and
Biology. Proceedings of the International Symposium on Plant Glyco-
sides, Kunming, China, C.-R. Yang, S.-H. Tanaka (eds). Elsevier Science:
Amsterdam, 1999; 373.
151. E. Galum, D. Avid, A. Dantes, A. Freeman. Planta Med. 1985, 51,
511.
152. G. Gil, P. F. dos Santos, C. Bullard. Phytochemistry 1995, 38, 629.
153. Y. X. Tang, T. Suga. Phytochemistry 1994, 37, 731.
154. I. Hook, R. Lecky, B. McKenna, H. Sheridan. Phytochemistry 1990, 29,
2143.
155. W. Zhu, G. B. Lockwood. Biotechnol. Lett. 2000, 22, 659.
156. W. Zhu, G. Asghari, G. B. Lockwood. Filoterapia 2000, 71, 501.
157. T. Mulder-Krieger, R. Verpoorte, A. Baerheim Svendsen, J. J. C. Schef-
fer. Plant Cell Tiss. Org. 1988, 13, 85.
158. A. A. Gbolade, G. B. Lockwood. J. Plant Physiol. 1990, 136, 198.
159. M. Lindmark-Henriksson, D. Isaksson, T. Vanek, I. Valterova, H. E.
Hogberg, K. Sjodin. J. Biotech. 2004, 107, 173.
160. T. Vanek, M. Novotny, R. Podlipna, D. Saman, I. Valterova. J. Nat. Prod.
2003, 66, 1239.
161. P. Pawlowicz, C. Wawrzenczyk, A. Siewinski. Phytochemistry 1992, 31,
2355.
162. N. Dudai, O. Larkov, E. Putievsky, H. R. Lerner, U. Ravid, E. Lewinsohn,
A. M. Mayer. Phytochemistry 2000, 55, 375.
163. L. G. Romagnoli, D. Knorr. Food Biotechnol. 1988, 2, 93.
164. D. Havkin-Frenkel, A. Podstolski. US Patent No. 7226783 B1, 2007.
383
View this article online at wileyonlinelibrary.com

165. C. Larroche, C. Creuly, J. B. Gros. Appl. Microbiol. Biotech. 1995, 43,
222.
166. M. Miyazawa, A. Sugawara. Nat. Prod. Res. 2006, 20, 731.
167. J. C. R. Demyttenaere, M. del Carmen Herrera, N. de Kimpe.
Phytochemistry 2000, 55, 363.
168. A. M. Humphrey, B. A. Skill, J. L. Kinderlerer. US Patent No. 6218158
B1, 2001.
169. A. Lomascolo, L. Lesage-Meesen, M. Haon, D. Navarro, C. Antona, C.
Faulds, A. Marcel. World J. Microb. Biotech. 2001, 17, 99.
170. L. Lesage-Meessen, A. Lomascolo, E. Bonnin, J-F. Thibault, A. Buleon,
M. Roller, M. Asther, E. Record, B. C. Ceccaldi, M. Asther. Appl.
Biochem. Biotechnol. 2002, 102, 141.
171. G. O. Buchanan, L. A. D. Williams, P. B. Reese. Phytochemistry 2000,
54, 39.
172. M. Furusawa, T. Hashimoto, Y. Noma, Y. Asakawa. Chem. Pharm. Bull.
2005, 53, 1513.
173. P. Huguency, B. Dumont, F. Ropert, J. M. Belin. In Proceedings of
Bioflavor ‘95, Dizon, France, 1995; 269.
174. K. Mitsuhashi, M. Iimori. US Patent No. 7129067, 2006.
175. K. Rüdiger, U. Krings, T. Nanzad, R. G. Berger. Appl. Microbiol. Biotech-
nol. 2005, 67, 477.
176. R. Cardillo, C. Fuganti, G. Sacerdote, M. Barbeni, P. Cabella, F.
Squarcia. US Patent No. 4950607, 1990.
177. D. O. Collins, G. O. Buchanan, W. F. Reynolds, P. B. Reese. Phytochem-
istry 2001, 57, 377.
178. Y. Noma, E. Akehi, N. Miki, Y. Asakawa. Phytochemistry 1992, 31,
515.
179. S. Gurdip, I. P. S. Kapoor, K. Jaspreet, O. P. Singh, S. K. Vernwal, R. S. S.
Yadav, G. P. Rao, S. R. Sharma, P. A. Leclercq, N. Klingby. J. Med.
Aromat. Plant Sci. 2002, 24, 23.
180. R. Kaspera, U. Krings, M. Pescheck, D. Shell, J. Schrader, R. G. Berger.
Z Naturforsch. C 2005, 60, 459.
181. B. H. de Oliveira, R. A. Strapasson. Braz. Arch. Biol. Technol. 2000, 43,
11.
182. D. Busmann, R. G. Berger. J. Biotech. 1994, 37, 39.
183. M. Miyazawa, Y. Honjo, H. Kameoka. Phytochemistry 1997, 44, 433.
184. H. Zorn, S. Langhoff, M. Scheibner, R. G. Berger. Appl. Microbiol.
Biotech. 2003, 62, 331.
185. M. M. W. Etschmann, J. Schrader. Appl. Microbiol. Biotechnol. 2006,
71, 440.
186. B. Kümin, T. Münch. European Patent No. EP 0795607, 1997.
187. B. Kumin, T. Munch. US Patent No. 5849551, 1988.
188. W. Wolken, M. van der Werf. Appl. Microbiol. Biotech. 2001, 57, 731.
189. C. Larroche, B. Tallu, J. B. Gros. J. Ind. Microbiol. 1988, 3, 1.
190. K. A. Jensen, K. M. C. Evans, T. Kent Kirk, K. E. Hammel. Appl. Environ
Microbiol. 1994, 60, 709.
191. W. D. Murray, S. G. B. Duff, P. H. Lanthier. US Patent No. 4871669,
1989.
192. J. Onken, R. G. Berger. J. Biotech. 1999, 69, 163.
193. T. Kawabe, H. Morita. J. Agric Food Chem. 1994, 42, 2556.
194. T. Kurniadi, R. B. Rhlid, L. B. Fay, M. A. Juillerat, R. G. Berger. J. Agric.
Food Chem. 2003, 51, 31-3-3107.
195. P. Marullo, C. Thibon, D. Dubourdieu, T. Takatoshi. European Patent
Application No. EP 1 927 654 A1, 2008.
196. M. I. Farbood, L. E. Kizer, J. Morris, G. Harris, L. B. McLean. US Patent
No. 6187741 B1, 2001.
197. D. Serp, U. von Stockar, I. W. Marison. Biotechnol. Bioeng. 2003, 82,
103.
198. D. Stark, T. Münch, B. Sonnleitner, I. W. Marison, U. von Stockar.
Biotechnol. Prog. 2002, 18, 514.
199. S. Maicas, J. J. Mateo. Appl. Microbiol. Biotech. 2005, 67, 322.
200. S.-L. Lee, H.-Y. Cheng, W.-C. Chen, C.-C. Chou. Process. Biochem. 1998,
33, 453.
201. L. Dufossé, G. Feron, G. Geneviève Mauvais, P. Bonnarme, A. Durand,
H. E. Spinnler. J. Ferment. Bioeng. 1998, 86, 169.
202. P. S. J. Cheetham. Trends Biotechnol. 1993, 11, 478.
203. A. Lomascolo, D. Asther, C. Navarro, C. Antona, M. Delattre, L.
Lesage-Meessen. Lett. Appl. Microbiol. 2001, 32, 262.
204. Y. Washé, M. Aguedo, A. Choquet, I. L. Gatfield, J.-M. Nicaud, J.-M.
Belin. Appl. Environ. Microbiol. 2001, 67, 5700.
205. Y. Waché, K. Bergmark, J. L. Courthaudon, M. Aguedo, J.-M. Nicaud,
J.-M. Belin. Lett. Appl. Microbiol. 2000, 30, 183.
206. U. Heyen, J. Harder. FEMS Microbiol. Lett. 1998, 169, 67.
207. J. Rabenhorst. In Biotechnology, Rehm H-J, Reed G (eds). Wiley VCH:
Weinheim, 2000; 8b, 333–350.
384
View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd.
Y. Gounaris
208. E. Pichersky. US Patent No. 5849526, 1998.
209. A. Sánchez-Contreras, M. Jiménez, S. Sanchez. Appl. Microbiol.
Biotech. 2000, 54, 528.
210. L.-Q. Zhao, Z.-H. Sun, P. Zheng, L. L. Zhu. Process. Biochem. 2006, 41,
1673.
211. C. Hockenmann, F. Juttner. Flavour Fragr. J. 2005, 20, 387.
212. S. Gocho, R. Kitazawa, T. Komal. European Patent No EP 0822259 A1.
1998.
213. U. Krings. Verfahren zur kontinuierlichen Gewinnung von Aromast-
offen aus dem Kulturmedium von Bioreactoren durch kontinuierli-
chen Festphasenextraktion. Dissertation, Universität Hannover,
1994.
214. T. Chatterjee. Biotech. Appl. Biochem. 2004, 39, 303.
215. M. J. van der Werf, P. M. Keijzer, P. H. van der Schaft. J. Biotech. 2000,
84, 133.
216. A. Boontawan, D. C. Stuckey. Appl. Microbiol. Biotech. 2006, 69,
643.
217. R. G. Berger. Aroma Biotechnology. Springer Verlag: Berlin, 1995.
218. I. M. Labuda, S. K. Goers, K. A. Keon. US Patent No. 5128253, 1992.
219. M. Yamada, Y. Okada, T. Yoshida, T. Nagasawa. Appl. Microbiol.
Biotech. 2007, 73, 1025.
220. P. Fontanille, A. Le Flèche, C. Larroche. Biocatal. Biotransform. 2002,
20, 413.
221. S. K. Yoo, D. F. Day, K. R. Cadwallader. Process. Biochem. 2001, 36,
925.
222. Z. Sun, P. Zheng, X. Guo, G. Lin, H. Yin, J. Wang, Y. Bai. European
Patent No. EP1734128 B1, 2009.
223. C. C. C. R. de Carvalho, M. M. R. da Fonseca. J. Mol. Catal. B 2002,
19–20C, 389.
224. C. C. C. R. de Carvalho, M. M. R. da Fonseca. Tetrahedron 2003, 14,
3925.
225. W. A. Duetz, A. H. M. Fjallman, S. Y. Ren, C. Jourdat, B. Witholt. Appl.
Environ. Microbiol. 2001, 67, 2829.
226. J. Rabenhorst, R. Hopp. US Patent No. 5017388, 1991.
227. J. A. Muheim, K. Lerch. Appl. Microbiol. Biol. 1999, 51, 456.
228. A. Muheim, B. Müller, T. Münch, M. Wetli. US Patent No. 6235507 B1,
2001.
229. P. Xu, D. Hua, S. Lin, L. Song, C. Ma, Z. Zhang, Y. Zeng, Y. Du, H. Chen,
L. Gan, Z. Wei. European Patent Application No. EP 2075327 A1,
2009.
230. L. M. Vane. J. Chem. Technol. Biotech. 2005, 80, 603.
231. F. Schade, J. E. Thompson, R. L. Legge. Biotech. Bioeng. 2003, 84,
265.
232. T. S. Johnson, G. A. Ravishankar. J. Plant Physiol. 1996, 147, 481.
233. N. Pras, H. J. Woerdenbag, W. van Uden. Agric. Biotechnol. News 1995,
7, 231.
234. P. Schreier. Adv. Biochem. Eng. Biotechnol. 1997, 55, 51.
235. B. J. Cass, F. Schade, C. W. Robinson, J. E. Thompson, R. L. Legge.
Biotech. Bioeng. 2000, 67, 372.
236. U. Kragl, W. Kruse, W. Hummel, C. Wandrey. Biotech. Bioeng. 1996,
52, 309.
237. D. M. de Temiño, W. Hartmeier, M. B. Ansorge-Schumacher. Enzyme
Microb. Technol. 2005, 36, 3.
238. A. Yoshida, Y. Takenaka, H. Tamaki, I. Frebort, O. Adachi, H. Kumagai.
J. Ferment. Bioeng. 1997, 84, 603.
239. R. Mikolajek, A. C. Spiess, J. Büchs. J. Biotech. 2007, 129, 723.
240. M. L. Fauconnier, J. Delcarte, M. Jaziri, P. D. U. Jardin, M. Marlier. J.
Plant Physiol. 2002, 159, 1055.
241. S. Lamare, R. Lortie, M. D. Legoy. Biotech. Bioeng. 1997, 56, 1.
242. D. P. Piet, R. Schrijvers, M. C. R. Franssen, E. De Groot. Tetrahedron
1995, 51, 6303.
243. M.-H. Lee, H. Liu, N.-W. Su, K.-L. Ku, Y.-M. Choong. J. Chin. Agric. Chem.
Soc. 1999, 37, 1.
244. J. W. de Kraker, M. Schurink, M. C. R. Franssen, W. A. König, A. de
Groot, H. Bouwmeester. Tetrahedron 2003, 59, 409.
245. A. Haeusler, K. Lerch, A. Muheim, N. Silke. US Patent No. 6238898,
2001.
246. A. Atwal, B. Bisakowski, S. Richard, N. Robert, B. Lee. Process. Biochem.
2005, 40, 95.
247. G. Bourel, J. M. Nicaud, B. Nthangeni, P. Santiago-Gomez, J. M. Belin,
F. Husson. Enz. Microb. Technol. 2004, 35, 293.
248. R. Huang, P. A. Christenson, I. M. Labuda. European Patent Applica-
tion No. EP1083233 A1, 2001.
249. A. Larios, H. S. Garsia, M. R. Oliart, M. R. Oliart, G. Valerio-Alfaro. Appl.
Microbiol. Biotechnol. 2004, 65, 373.
Flavour Fragr. J. 2010, 25, 367–386
Biotechnology for essential oils, flavours and volatile isolates
250. S. Srivastava, J. Modak, G. Madras. Ind. Eng. Chem. Res. 2002, 41,
1940.
251. G. A. Makedo, M. M. S. Lozano, G. M. Pastore. Electron. J. Biotechnol.
2003, 6, 72.
252. A. M. Almosnino, M. Bensoussan, J. M. Belin. Food Chem. 1996, 55,
327.
253. M. A. Noordermeer, W. van der Goot, A. J. van Kool, J. W. Veldsink,
G. A. Veldink, J. F. G. Vliegenthart. J. Agric. Food Chem. 2002, 50,
4270.
254. M.-R. Kula, U. Kragle. In Stereoselective Biocatalysis, R. N. Patel (ed.).
Markel Dekker: New York, 2000; 839.
255. F. Ruiz-Terán, I. Perez-Amador, A. López-Munguia. J. Agric. Food
Chem. 2001, 49, 5207.
256. J. Kaminska, L. Marcowicz, J. Stołowska, J. Gòra. Enzyme Microbiol.
Biotech. 1989, 11, 436.
257. N. Pras, G. E. Booi, D. Dijkstra, A. S. Horn, T. M. Malingre. Plant Cell Tiss.
Org. 1990, 21, 9.
258. C. Dulieu, M. Moll, J. Boudrant, D. Poncelet. Biotechnol. Progr. 2000,
16, 958.
259. Z. Gunata, C. Bayonove, C. Tapiero, R. Cordonnier. J. Agric. Food
Chem. 1990, 38, 1232.
260. Y. Vasserot, A. Arnaud, P. Galzy. Acta Biotechnol. 1995, 15, 77.
261. H. J. Bouwmeester, J.-W. de Kraker, M. Schurink, R. J. Bino, A. de Groot
A, M. C. R. Franssen. US Patent No. 7214507 B2, 2007.
262. B. Muller, A. Gautier, C. Dean, J.-C. Kuhn. US Patent No. 5464761,
1995.
263. J. Memelink, J. W. Kijne, R. van der Heijden, R. Verpoorte. Adv.
Biochem. Engin. Biotechnol. 2001, 72, 103.
264. B. N. Mijts, C. Schmidt-Dannert. Curr. Opin. Biotechnol. 2003, 14, 597.
265. R. Verpoorte, J. Memelink. Curr. Opin. Biotechnol. 2002, 13, 181.
266. R. Verpoorte, R. van der Heijden, H. J. G. ten Hoopen, J. Memelink.
Biotechnol. Lett. 1999, 21, 467.
267. A. Aharoni, M. A. Jongsma, H. J. Bouwmeester. Trends Plant Sci. 2005,
10, 504.
268. A. Aharoni, M. A. Jongsma, T.-Y. Kim, M.-B. Ri, A. P. Giri, F. W. A.
Verstappen, W. Schwab, H. J. Bouwmeester. Phytochemistry Rev.
2006, 5, 49.
269. M. R. Wildung, R. B. Croteau. Transg. Res. 2005, 14, 365.
270. K. Reiling, Y. Yoshikuni, V. J. J. Martin, J. Newman, J. Bohlmann, J. D.
Keasling. Biotech. Bioeng. 2004, 87, 200.
271. J. W. Frost. US Patent No. 6372461 B1, 2002.
272. J. D. Keasling, V. J. J. Martin, D. J. Pitera, S.-W. Kim, S. T. Withers III, Y.
Yoshikuni, J. Newman, A. V. Khlebnikov. US Patent No. 7622282 B2,
2009.
273. C. Ohto, S. Obata, M. Muramatsu, K. Nishi, K. Totsuka. US Patent No.
7501268 B2, 2009.
274. M. Schalk. US Patent No. 7622288 B2, 2009.
275. P. Barghini, D. Di Gioia, F. Fava, M. Ruzzi. Microb. Cell Fact. 2007, 6,
13.
276. P. J. Oriel, S. Savithiry, H. C. Chang. US Patent No. 5688673, 1997.
277. N. Savithiry, P. J. Oriel. US Patent No. 5763237, 1998.
278. D. J. DiCosimo, M. Koffas, J. M. Odom, S. Wang. US Patent No.
6818424 B2, 2004.
279. E. A. Soher, N. Abo-Serih, K. F. El Massry. Arab Univ. J. Agric. Sci. 2003,
11, 205.
280. B. E. Jackson, E. A. Hart-Wells, S. P. T. Matsuda. Org. Lett. 2003, 5,
1629.
281. S. P. T. Matsuda, E. A. Hart. US Patent No. 7238514 B2, 2007.
282. J. D. Keasling, Y. Shiba, J. Kirby, E. M. Paradise. US Patent No. 0053797,
2009.
283. H. Tsuruta, J. R. Lenihan, R. Regentin. US Patent No. 0137014,
2009.
284. A. Aharoni, M. A. Jongsma, H. J. Bouwmeester. US Patent No.
0031455, 2009.
285. Y. Waché, M. Aguedo, A. Choquet, I. L. Gatfield, J.-M. Nicaud, J.-M.
Belin. Appl. Environ. Microbiol. 2001, 67, 5700.
286. Y. Pagot, A. Endrizzi, J. M. Nicaud, J.-M. Belin. Lett. Appl. Microbiol.
1997, 25, 113.
287. A. Groguenin, Y. Waché, E. E. Escamilla Garcia, M. Aguedo, F. Husson,
M. T. LeDall, J.-M. Nicaud, J.-M. Belin. J. Mol. Catal. 2004, 28, 75.
288. Y. Waché, C. Laroche, K. Bergmark, C. Møller-Andersen, M. Aguedo,
M.-T. Le Dall, H. Wang, J.-M. Nicaud, J.-M. Belin. Appl. Environ. Micro-
biol. 2000, 66, 1233.
289. Y. Waché, M. Aguedo, M.-T. LeDall, J.-M. Nicaud, J.-M. Belin. J. Mol.
Catal. B Enzym. 2002, 19–20, 347.
Flavour Fragr. J. 2010, 25, 367–386 Copyright © 2010 John Wiley & Sons, Ltd.
290. D. P. Mobley, G. K. Shank. World Intellectual Property Organization
Patent No. WO0017380, 2000.
291. Y. Zhang, R. C. Wilson, D. L. Craft, D. L. Eirich, R. W. Frayer. World
Intellectual Property Organization Patent No. WO2004013336,
2004.
292. T. Zimmer, K. Kaminski, W.-H. Schunck, E. Kärgel, U. Scheller, S.
Mauersberger. World Intellectual Property Organization Patent No.
WO9627678, 1996.
293. S. S. Mahmoud, R. B. Croteau. Proc. Natl Acad. Sci. USA 2001, 98, 8915.
294. S. Krasnyanski, R. A. May, A. Loskutov, T. M. Ball, K. C. Sink. Theor.
Appl. Genet. 1999, 99, 676.
295. S. S. Mahmoud, M. Williams, R. Croteau. Phytochemistry 2004, 65,
547.
296. F. Diemer, J. C. Caissard, S. Moja, J. C. Chalchat, F. Jullien. Plant Physiol.
Biochem. 2001, 39, 603.
297. K. Ohara, T. Ujihara, T. Endo, F. Sato, K. Yazaki. J. Exp. Bot. 2003, 54,
2635.
298. J. Lücker, W. Schwab, M. C. R. Franssen, L. H. W. van der Plas, H. J.
Bouwmeester, H. A. Verhoeven. Plant J. 2004, 39, 135.
299. M. Lavy, A. Zuker, E. Lewinsohn, O. Larkov, U. Ravid, A. Vainstein, D.
Weiss. Mol. Breeding 2002, 9, 103.
300. E. Lewinsohn, F. Schalechet, J. Wilkinsohn, K. Matsui, Y. Tadmor, K. H.
Nam, O. Amar, E. Lastochkin, O. Larkov, U. Ravid, W. Hiatt, S. Gep-
stein, E. Picherski. Plant Physiol. 2001, 127, 1256.
301. J. Lücker, H. J. Bouwmeester, W. Schwab. Plant J. 2001, 27, 315.
302. A. Aharoni, A. P. Giri, S. Deuerlein, F. Griepink, W. J. deKogel, F. W. A.
Verstappen, H. A. Verhoeven, M. A. Jongsma, W. Schwab, H. J. Bou-
wmeester. Plant Cell 2003, 15, 2866.
303. A. Aharoni, M. A. Jongsma, H. A. Verhoeven, H. J. Bouwmeester. US
Patent No. 7453024 B2, 2008.
304. S. S. Mahmoud, R. G. Croteau. Proc. Natl Acad. Sci. USA 2003, 100,
14481.
305. I. F. Kappers, A. T. Aharoni, W. J. M. van Herpen, L. L. P. Luckerhoff, M.
Dicke, H. J. Bouwmeester. Science 2005, 309, 2070.
306. C. Schnee, T. G. Kollner, M. Held, T. C. J. Turlings, J. Gershenzon, J.
Degenhardt. Proc. Natl Acad. Sci. USA 2006, 103, 1129.
307. T. M. Hohn, J. B. Ohlrogge. Plant Physiol. 1991, 97, 460.
308. M. H. Beale, M. A. Birkett, T. J. A. Bruce, K. Chamberlain, L. M. Field,
A. K. Huttly, J. L. Martin, R. Parker, A. L. Phillips, J. A. Pickett, I. M.
Prosser, P. R. Shewry, L. E. Smart. Proc. Natl Acad. Sci. USA 2006, 103,
10509.
309. I. Guterman, T. Masci, X. Chen, F. Negre, E. Pichersky, N. Dudareva, D.
Weiss, A. Vainstein. Plant Mol. Biol. 2006, 60, 555.
310. C. Tesniere, L. Torregrosa, M. Pradal, J. M. Souquet, C. Gilles, K. dos
Santos, P. Chatelet, Z. Gunata. J. Exp. Bot. 2006, 57, 91.
311. D. Tieman, M. Taylor, N. Schauer, A. R. Fernie, A. D. Hanson, H. J. Klee.
Proc. Natl Acad. Sci. USA 2006, 103, 8287.
312. B. A. Underwood, D. M. Tieman, K. Shibuya, R. J. Dexter, H. M. Loucas,
A. J. Shimkin, C. A. Sims, E. A. Schmelz, H. J. Klee, D. J. Clark. Plant
Physiol. 2005, 138, 255.
313. A. Zuker, T. Tzfira, H. Ben-Meir, M. Ovadis, E. Shklarman, H. Itzhaki, G.
Forkmann, S. Martens, I. Neta-Sharir, D. Weiss, A. Vainstein. Mol.
Breeding 2002, 9, 33.
314. J. C. Verdonk, M. A. Haring, A. J. van Tunen, R. C. Schuuring. Plant
Cell 2005, 17, 1612.
315. J. Boatright, F. Negre, X. Chen, C. M. Kish, B. Wood, G. Peel, I.
Orlova, D. Gang, D. Rhodes, N. Dudareva. Plant Physiol. 2004, 135,
1993.
316. H. Y. Gao, B. Z. Zhu, H. L. Zhu, Y. L. Zhang, Y. H. Xie, Y. C. Li, Y. B. Luo.
Russ. J. Plant Physiol. 2007, 54, 80.
317. A. M. Dandekar, J. R. Stow, R. J. Colgan, D. J. James. Transg. Res. 2004,
13, 373.
318. B. G. Defillipi, A. M. Dandekar, A. A. Kader. J. Agric. Food Chem. 2004,
52, 5694.
319. B. G. Defillipi, A. A. Kader, A. M. Dandekar. Plant Sci. 2005, 168, 1199.
320. A. D. Bauchot, D. S. Mottram, A. T. Dotson, P. John. J. Agric. Food Chem.
1998, 46, 4787.
321. F. Flores, F. El Yahyaoui, G. de Billerbeck, F. Romojaro, A. Latche, M.
Bouzayen, J. C. Pech, C. Ambid. J. Exp. Bot. 2002, 53, 201.
322. C. Wang, J. Xing, C. K. Chin, C. T. Ho, C. E. Martin. Phytochemistry
2001, 58, 227.
323. M. Hong, B. A. Zilinskas, D. C. Knipple, C. K. Chin. Phytochemistry
2004, 65, 159.
324. J. Speirs, E. Lee, K. Holt, Y-D. Kim, N. S. Scott, B. Loveys, W. Schuch.
Plant Physiol. 1998, 117, 1047.
385
View this article online at wileyonlinelibrary.com

325. S. Prestage, R. S. T. Linforth, A. J. Taylor, E. Lee, J. Speirs, W. Schuch.
J. Sci. Food Agric. 1999, 79, 131.
326. E. W. Chehab, G. Raman, J. W. Walley, J. V. Perea, G. Banu, S. Theg, K.
Dehesh. Plant Physiol. 2006, 141, 121.
327. K. Matsui, S. Fukutomi, J. Wilkinson, B. Hiatt, V. Knauf, T. Kajwara.
J. Agric. Food Chem. 2001, 49, 5418.
328. G. Vancanneyt, C. Sanz, T. Farmaki, M. Paneque, F. Ortego, P.
Castanera, J. J. Sanchez-Serrano. Proc. Natl Acad. Sci. USA 2001, 98,
8139.
386
View this article online at wileyonlinelibrary.com Copyright © 2010 John Wiley & Sons, Ltd.
Y. Gounaris
329. G.-P. Chen, R. Hackett, D. Walker, A. Taylor, Z-F. Lin, D. Grierson. Plant
Physiol. 2004, 136, 2641.
330. W. L. Deng, W. S. Grayburn, T. R. Hamilton-Kemp, G. B. Collins, D. F.
Hildebrand. Planta 1992, 187, 203.
331. J. Leon, J. Royo, G. Vancanneyt, C. Sanz, H. Silkowski, G. Griffiths, J. J.
Sanchez-Serrana. J. Biol. Chem. 2002, 277, 416.
332. H. J. Klee, D. Tieman. World Intellectual Property Organization
Patent No. WO 2005/035752 A2, 2005.
333. S.-H. Kim, H.-C. Ji, K.-B. Lim. J. Plant Biotechnol. 2005, 7, 87.
Flavour Fragr. J. 2010, 25, 367–386