Indian Journal of Biotechnology

Vol 8, July 2009, pp 304-310

Influence of Pediococcus acidilactici as a starter on the flavour of tempoyak

(fermented durian)
Neti Yuliana1* and Virgilio V Garcia2
1
Jurusan Teknologi Hasil Pertanian (THP), Fakultas Pertanian,
Universitas Lampung (UNILA). Jalan Sumantri Brojonegoro #1 Bandar Lampung, Indonesia
2
Institute of Food Science and Technology, University of the Philippine at Los Banos, Philippines
Received 24 July 2008; revised 28 November 2008; accepted 10 February 2009

Pediococcus acidilactici UP02 was used as a starter for the fermentation of tempoyak (Durio zibethinus Murr.), a
traditional Indonesian fermented fruit. The flavour of tempoyak was compared to those in spontaneous fermentation (no
starter added). The results showed that addition of P. acidilactici UP02 in tempoyak fermentation decreased the quantity of
sulphur components, reduced the sugar content, and increased various non-sulphur compounds as well as, non-volatile
acidity and contents of organic acids (lactic, malic, and acetic). Sensory evaluation showed that there was no significant
difference among the samples for sourness and aroma. However, the P. acidilactici UP02-inoculated sample was more
acceptable than the sample with no-starter treatment. On the basis of the results of GC-MS, the flavour components of
tempoyak with P. acidilactici UP02 treatment were observed to have 48 compounds, mainly composed of dimethyl disulfide
and diethyl disulfide followed by ethane,1-1-bis ethylthio.

Keywords: Pediococcus acidilactici, tempoyak, malic acid, sulphur components

Introduction final pH of 3.8 to 4.6 whereas, the acidity content of

Tempoyak (Durio zibethinus Murr.) is a fermented
durian, which is a popular side dish and condiment in
Malaysia and Indonesia, mainly in Sumatra and
Kalimantan islands1,2. This product is creamy, yellow-
white in colour, and has a distinctive aroma. It is
prepared by placing durian pulp in jars to which salt
(2-15%) is added and carefully mixed. The jars are
then tightly closed and kept for 7 d to let spontaneous
fermentation to take place3,4. During fermentation, the
texture of the durian pulp changes from solid to a
semisolid mass accompanied by a strong flavour; the
flavour is the result of a unique combination of
various sugars, organic acids as well as various
volatile organic compounds. The volatile organic
compounds in tempoyak consist of sulphur-containing
compounds; 3,5-dimethyl-1,2,4 trithiolane being the
highest, and non-sulphur compounds with butanoic
acid, 2-methyl ethyl ester being the highest4. Three
organic acids include acetic, lactic and malic; malic
being the most abundant5. These organic acids not
only contribute to taste compounds, but also increase
the acidity of tempoyak. The acidity of tempoyak
produced in Malaysia is 3.6% (as acetic acid) with a
________________
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E-mail: netiyuliana@yahoo.com
tempoyak from Indonesia is 2.55 and 2.56% (as lactic
acid)6-8. This high acidity of tempoyak is attributed to
lactic acid bacteria (LAB)4,9,10. Several researchers
have isolated many species of LAB from tempoyak.
Lactobacillus plantarum, L. brevis, L. mali and L.
fermentum have been found in Malaysian
tempoyak10,11, whereas L. plantarum, L. casei and L.
corynebacterium have been found in tempoyak from
Indonesia12. A new species, Lactobacillus durianis,
isolated from Malaysian tempoyak, has also been
reported13. Other LAB found in tempoyak are
Leuconostoc mesenteroides12, Pediococcus
4
acidilactici and Weissella mesenteroides . Historically
the species of Lactobacillus, Leuconostoc,
Pediococcus, and Streptococcus are the main species
of LAB.
There are numerous examples of controlled
fermentation using LAB as a starter for the processing
of foods, such as, green olive, fermented soymilk, and
minced mackerel production14-16. The use of a starter
culture provides consistency and reliability of
performance. Reports on the LAB as starters for
tempoyak are limited. Fermentation, using starters,
may affect the flavour of tempoyak. LAB can vary
metabolite reactions that produce various
17-19
compounds . These compounds are important as
 YULIANA & GARCIA: INFLUENCE OF P. ACIDILACTICI ON FLAVOUR OF TEMPOYAK
they impart characteristic taste and aroma to the final
product. The objective of this research was to evaluate
the flavour in tempoyak using P. acidilactici UP02 as
a starter, which has homofermentative nature and its
origin from tempoyak has been reported in a previous
study.
Materials and Methods
Preparation of Inoculum
Pediococcus acidilactici UP02 was obtained from
the Institute of Food Science and Technology,
University of the Philippines, Los Banos, Philippines.
This isolate was cultured and sub-cultured
anaerobically in the MRS medium (Difco, USA). The
cultures were centrifuged for 15 min at 3000 rpm and
resuspended in saline solution (0.85% NaCl) to yield
108CFU/mL for a working starter.
Preparation of Tempoyak
The flesh of the durians was aseptically taken and
divided into two sets of four representative treatments
comprising both with and without starter, and the
number of fermentation days (0,4,8 and 12). Two
replications were maintained in this experiment. Salt
was added to each treatment at the rate of 3% (w/w),
based on the durian flesh weight, and each lot (200 g)
was packed in a plastic container. One lot was
inoculated with P. acidilactici UP02 and the other lot
was naturally fermented (without adding starter). The
inoculation rate was 1%, based on the weight of
durian. The samples were then kept in a cabinet at
room temperature (27±1oC) for evaluation. Samples
from each treatment were withdrawn at 0, 4, 8, and 12
d fermentation for the analysis of total volatile acids,
non-volatile acids and reducing sugars. Data were
calculated as mean of two replications.
Analysis of Tempoyak
Procedures described by AOAC20 were used for the
examination of total volatile and non-volatile acids,
whereas reducing sugars were determined by Nelson-
Samogyi method21. Evaluation of flavour compounds
was done using the solid phase micro extraction
(SPME) trapping method and the quantification of
individual organic acids by HPLC22 as well as sensory
analysis was done on the 8 d fermented samples. To
assess the development of flavour during tempoyak
fermentation, sensory evaluation on the aroma and
sourness was performed by scoring test, although
those on the general acceptability were performed by
the hedonic test.
305
Evaluation of Individual Organic Acids
A 20 µL of tempoyak sample was injected after
filtering through a 0.45 µm membrane filter in a
Seppack C-18 cartridge (Waters Associate, USA).
HPLC conditions were as follows: Merck Polysphere
OA HY column (6 mm × 300 mm); mobile phase 0.01
N H2SO4; flow rate 0.8 mL/min. The peak areas of
each organic acid in the samples were calculated from
the integrator system of the HPLC data station.
Organic acid standards used were lactic, malic, acetic,
succinic and citric.
Determination of Total Volatile Acid Content
To an approximately 10 g homogenized sample
contained in a distilling flask, 100 mL of distilled
water was added. Distillation was conducted rapidly
until 100 mL of distillate was collected. The distillate
was titrated when still hot with standard 0.1 N NaOH,
using phenolphthalein as an indicator. The results
have been expressed as mg malic acid/g sample (wet
basis).
Determination of Total Non-volatile Acid Content
A 25 mL of sample (15 g diluted to 150 mL) was
evaporated to dryness in an evaporating dish on a
steam bath and dried for 30 min in an oven at 100oC.
The residue was dissolved in 25 mL of distilled water
and titrated with 0.1N NaOH using phenolphthalein
as an indicator. The amount of non-volatile acids was
calculated in terms of mL malic acid/g sample (wet
basis).
Evaluation of Volatile Flavour Compounds
A 75 g of sample was put into an Erlenmeyer flask,
sealed with aluminum foil with a protective seal, and
fitted with an solid phase microextraction (SPME) gas
tight syringe in such a way that the syringe would be
just above the food sample. The flask was placed in a
water bath at 50oC with the syringe still fitted. The
volatiles released from the sample during 30 min
period at 50oC were swept onto the SPME. These were
then analyzed using an HP 5890A gas chromatograph
connected to an HP 5970 mass spectrometer (Hewlett-
Packard). Gas chromatography-mass spectrometry
(GC-MS) was operated at 70 eV in the electron impact
(EI) mode over the range 35-450 amu; column used
was SPB-5; 30 m ×0.25 mm, film thickness = 0.25 µm,
(Supelco, Sigma-Aldrich Co.). The carrier gas was
helium at a flow rate of 1 mL/min. The collected
volatiles were thermally desorbed at 250oC for 2 min
after desorption. The oven was heated rapidly to 60oC
and maintained at this temperature for 2 min before the
306 INDIAN J BIOTECHNOL, JULY 2009

temperature was increased at 10oC min-1 to 220oC (10 as homofermentative24. However, the result of this
min). The constituents of samples were identified by study showed that malic acid was higher than lactic
matching their mass spectra with those recorded in the acid. The homofermentative LAB produced mainly
computer library (NIST98 and Wiley library). lactic acid from sugar metabolism, but significant
Evaluation of Reducing Sugars
changes in product formation could occur with sugar
Reducing sugars were determined by the Nelson- limitation and aeration. Co-metabolism of sugars and
Samogyi method. To 1 mL of the sample, 1 mL of a organic acids was common and often led to the
freshly prepared mixture of 25 parts copper reagent A production of a different product18.
to 1 part copper reagent B was added. It was mixed The organic acids present in tempoyak were not
and heated in a boiling water bath for 20 min and identified in the previous studies. The advantage of
rapidly cooled. To each tube of the mixture, 1 mL of this finding is that lactic acid and acetic acid contents
arsenomolybdate reagent was added to stabilize the were individually investigated using HPLC that
colour. The absorbance of sample at 500 nm was differed from others. Most authors7-9 reported both
measured using a spectrophotometer (Spectronic 20). acids as the total acidity through titration method. The
The value generated was compared with a standard total acidity of tempoyak was expressed as acetic
curve, to get the amount of reducing sugars in the (3.6%) and lactic acids (2.55-2.56%)6,8.
sample. Total Volatile and Non-volatile Acid Content
Sensory Evaluation of Flavour Developed during Tempoyak Table 1 shows that the non-volatile acidity value of
Fermentation through Scoring and Hedonic Tests starter and no-starter treatments at the start of
Samples of tempoyak with and without starter were fermentation was 3.55 mg malic acid/g sample. On
subjected to an 18-member panel for aroma, sourness, day-4 of the fermentation, the non-volatile acidity
and acceptability evaluation through scoring and values increased significantly to 9.72 and 13.27 mg
hedonic tests. Panel members consisted of Indonesian malic acid/g sample for no-starter and starter
students (male and female) who were familiar with treatments, respectively. This trend was also observed
durian and tempoyak. Data were analyzed by for volatile acidity.
ANOVA and Duncan’s Multiple Range Test at 5% The non-volatile acidity had the highest values on
level of significance 23. day-four of fermentation and was significantly different
in all treatments. Samples with P. acidilactici UP02
Results and Discussion treatment had the highest values of non-volatile acidity,
Organic Acid Contents followed by the no-starter treatment. After the day-4
Five organic acids (lactic, malic, acetic, succinic fermentation, the non-volatile acidity values of all the
and citric) were analyzed. However, citric and
succinic acids were not found in both the starter and Table 1—Organic acids identified and changes in volatile acidity
non-starter treated samples of tempoyak. The most and non volatile acidity in tempoyak inoculated with and without
Pediococcus acidilactici UP02
abundant organic acid found in tempoyak with P.
acidilactici UP02 starter was malic acid (14.60 Organic acids Fermentation No starter With
mg/mL) followed by lactic acid (4.34 mg/mL) and a (day) Pediococcus
small amount of acetic acid (1.63 mg/mL). Addition Malic 14.60±0.02 18.17±0.06
of the starter, however, showed an increase in the Lactic 3.41±0.03 4.34±0.06
amount of organic acids. This finding indicated that a Acetic 1.42±0.01 1.63±0.08
species of LAB had their own particular reactions and (mg/mL sample)
could vary with the conditions of fermentation. Non-volatile acid 0 3.55±0.09 3.55 ±0.09
malic (mg/g sample) 4 9.72±0.09 13.27±0.19
Succinic and citric acids were reported as common
8 9.38±0.00 12.97±0.15
non-volatile acids in one of the LAB products such as 12 9.45±0.09 12.93±0.09
Korean kimchi (Dongchimi radish), along with malic, Volatile acid acetic 0 0.24±0.00 0.24±0.00
lactic and acetic acids22. The conversion of sugars to (mg/g sample) 4 2.76±0.34 0.57±0.04
organic acids was influenced by the kind of LAB and 8 2.79±0.04 0.63±0.04
its ability to ferment carbohydrates. Production of 12 2.82±0.08 0.65±0.02
lactic acid was a common characteristic in Values are expressed as mean ± standard deviation of two
homofermentatives3 and P. acidilactici was grouped determinations
 YULIANA & GARCIA: INFLUENCE OF P. ACIDILACTICI ON FLAVOUR OF TEMPOYAK 307

samples gradually decreased (Table 1). This was 0.33%; the highest value reached after 21 d of
probably because of the initial reaction of the acids fermentation25. The volatile acidity in tempoyak was
with alcohol in the formation of other metabolites. also probably contributed by octanoic, ethane dithioic,
Volatile acid value of all the samples, as shown in hexanoic and ethylthiobutanoic acids, besides acetic
Table 1, at the beginning of fermentation, was 0.24 acid, as analyzed by GC-MS4.
acetic acid/g. It was observed that the volatile acidity
values of both treatments increased significantly on Volatile Flavour Compounds
day-4 of fermentation (0.63-2.79 mg acetic acid/g). The analysis of an 8-d-old sample of tempoyak
Samples with P. acidilactici UP02 starter had low inoculated with and without P. acidilactici UP02 for
volatile acidity value (0.63 mg acetic acid/g, whereas, volatile sulphur compounds and fruity flavour
the no-starter treatment had a high volatile acidity component showed that there were distinct
value (2.79 mg acetic acid/g), equivalent to 0.28%. differences in the number of flavour components
This value was not so different than for the volatile (Tables 2 & 3). The flavour components of tempoyak
acid reported in fermented cabbage, which was around with P. acidilactici UP02 treatment were observed to
have 48 compounds whereas no-starter treatment had
Table 2Sulphur components of tempoyak inculated with 22 compounds. These compounds corresponded to
without P. acidilactici thiol, ester, alcohols, aldehyde, alkane, ketone, and
No Components Area% small amounts of unknown substances.
No With All the flavour compounds produced were
Starter Pediococcus dominated by sulphur compound, diethyl tyrisulfide.
1 Disulfide,ethyl (1- * 0.38 This did not confirm with the pervious result4, where
methylprophyl)(I) 3,5-dimethyl-1, 2,4-trithiolane was reported to be
2 Methyl ethyldisulfide * 0.51 present in highest concentration in naturally
3 Unknown (S compound) 1.41 * fermented tempoyak. This difference could be
4 Disulfide,diethyl 13.44 9.8 attributed to the different variety used and maturity of
5 Ethylthio acetate * 0.33 durian as well as the different microorganism
6 Unknown (S compound) * 0.88 combinations involved.
7 1,2,4-Trithiolane, 7.02 4.39 Analysis also showed that both starter and no-
3,5-dimethyl starter treatment sample components had less ethyl 2-
8 Diethyltrisulfide 39.68 34.25 methyl butanoate, indicating that the samples had a
9 3-Ethyl,5-methyl-1,2,4-trithiolane * 0.04 less fruity flavour constituting the durian flavour.
(I) Durian fruits possess two distinct odor notes, (i) a
10 2-(Methylthio) ethanol 6.5 6.82 strong sulfury onion-like and (ii) a delicately fruity26.
11 3-Ethyl,5-methyl-1,2,4-trithiolane * 0.16 It had been observed in the previous reports that ethyl
(II) 2-methyl butanoate imparted the fruity durian
12 Ethyl n-prophyl disulfide 1.83 3.21 flavour26,27. Moreover, it has been found that this
13 3-Ethylthiobutanoic acid 0.67 * compound contributed most to the non-sulphur part of
14 Unknown (S compound) 0.52 * the durian flavour as analyzed by GC-Sniff flavour
15 Ethane,1-1-bis(methylthio) 1.66 1.1 dilution analysis28.
16 Ethane,1-1-bis(ethylthio)(I) 13.69 7.92 The 48 volatile compounds in the sample with P.
17 Trisulfide,diprophyl * 0.26 acidilactici UP02 were characterized by the presence
Disulfide,ethyl (1-methyl) prophyl of 18 sulphur, 10 ester, 2 carboxylic acid, 5 alcohol, 2
18 (II) * 0.17 aldeyhe, 2 alkane, 1 ketone, 6 unknown, and 2
N-dimethylthiophosphynil-3-amino miscellaneous compounds. Many compounds
19 pyridine * 8.5 generated in this sample were not found in
4,6-Dimethyl-1,2,3,5- the no-starter added samples, such as acetic acid;
20 tetrathiacycloghexane * 0.26 ethylthio ethane; ethylpropionate; hexanoic acid; 2,3-
21 Methanethioamide,N,N-dimethyl- * 0.5 dihydro-benzofuran; 1-acetylamino-5,5-dimethoxy-
22 3-(2-Mercapto-methyl prophyl)thio 1.65 * 2,3-dimethyl 2 pentene; germacyclobutane,1,1-
* not found dimethyl; disulfide,ethyl (1 methylpriophyl); N-dim-
308 INDIAN J BIOTECHNOL, JULY 2009

Table 3—Non-sulphur components of tempoyak inoculated with ioamide,N,N-dimethyl and glycine methyl ester. The
and without P. acidilactici last compound was the major constituent of the non-
No Components Area (%) sulphur volatiles present in the samples inoculated by
No Starter With P. acidilactici UP02 starter.
Pediococcus Among the sulphur compounds, disulfide diethyl
1 Acetic acid
was the second abundant sulphur compound observed
* 1.24
2 Ethyl propionate
in all the samples with and without starter treatment.
* 0.21
3 Butanone, 3-hydroxyl
This may be responsible for the fetid odour that was
* 0.3
4 Unknown
noted in fermented durian. Disulfide diethyl is
0.59 *
reported to be responsible for the fetid odour in
5 1-Butanol, 3methyl 0.69 0.51
durian29. As much as 18 sulphur compounds were
6 1 -Butanol, 2-methyl- * 0.14
detected in P. acidilactici UP02 treated samples,
7 2,3-Butanediol 1.93 0.66
whereas no-starter treated showed 10 sulphur
8 1,3-Butanediol 2.14 0.75 compounds. Presence of abundant sulphur
9 Butanoic acid ethyl ester * 0.29 components caused the samples to still have a strong
10 Propanoic acid, 2-hydroxy,ethyl
ester * 0.36 onion-like odour.
11 2-Butenoic acid, ethyl ester * 0.25 Head space analysis of no-starter treated samples
12 Butanoic acid, 2 methyl, ethyl revealed that 1-butanol 3-methyl; 2,3-butanediol; 1,3-
ester 0.49 0.46 butanediol; butanoic acid, 2-methyl, ethyl ester;
13 Unknown * 0.16 nonanoic acid, ethyl ester; and two unknown
14 Unknown * 0.79 compounds were the principal non-sulphur aromas
15 Butanoic acid, 3-hydroxy,ethyl present. Among these existing non-sulphur
ester * 0.35
16 Unknown
compounds, 1,3-butanediol was found in highest
0.46 *
17 Hexanoic acid
amount in thie samples. It was noticed that nonanoic
* 0.46
18 Hexanoic acid, ethyl ester
acid, ethyl ester, and the sulphur compound, 3-
* 0.45
19 Unknown
ethylthiobutanoic acid were only found in the no-
0.93 0.97
starter treated samples.
20 1-Octanol * 0.34
21 Nonyl aldehide * 0.3 Reducing Sugars
22 Unknown * 0.24 The results showed that starter and no-starter
23 4-(1-Hydroxy-ethyl) gamma treatments had similar patterns of reducing sugars,
butanolacton * 0.2 whose values decreased from day-0 to day-8 of
24 Ethyl octanoate/ethyl caprylate * 1.05 fermentation period (Fig. 1). This was due to the
25 2,3-Dihydro-
benzofuran/acetophenon * 0.12
utilization of low molecular weight reducing sugars as
26 Nonanoic acid, ethyl ester 1.47 *
a source of energy by the LAB.
27 Unknown 0.5 *
Higher reducing sugars content, however, was
28 2 Decenal * 0.25
observed in the no-starter added treatment rather than in
29 2-pentene * 0.25
the P. acidilactici UP02 treated tempoyak. The values
30 Germacyclobutane,1,1-dimethyl * 0.19
generated in the experiment might be caused by the
31 Unknown 1.36 *
presence of other microflora at initial fermentation,
32 Glycine, methyl ester besides LAB. Many microflora such as fungi were able
* 3.04
33 Unknown to produce enzymes, which in turn, degraded
* 1.28
34 lauric acid, ethyl ester carbohydrates to simple sugars such as glucose and
* 0.14
35 Unknown fructose. The aril of durian is known to contain about
0.88 *
36 Tetradecanoic acid, ethyl ester 33% carbohydrates on a fresh weight basis, of which
* 0.1
37 Hexadecanoic acid, ethyl ester about one third is probably starch30. In the P. acidilactici
* 0.35
* not found
UP02 treatment, the decreasing value of reducing
sugars, in this experiment, was probably due to the high
ethylthiophosphynil-3- aminopyridine: tetradecanoic rate of consumption by the LAB during fermentation. P.
acid ethyl ester; hexadecanoic acid ethyl ester; 4,6 acidilactici UP02 was present in high amounts since the
dimethyl-1,2,3,5,5-tetrathiacyclohexane; methaneth- beginning of fermentation. The growth kinetics of
 YULIANA & GARCIA: INFLUENCE OF P. ACIDILACTICI ON FLAVOUR OF TEMPOYAK
Fig. 1—Changes in reducing sugars (%) (wet basis) during
fermentation of tempoyak with and without P. acidilactici.
Table 4—Sensory evaluation of tempoyak inoculated with and
without Pediococcus acidilactici
Parameters Treatments
Pediococcus No-starter added
Sourness 3.1a 3.6ab
Aroma 3.3a 3.4a
General acceptability 3.6a 4.13b
microorganisms present in the samples determined the
consumption rate of reducing sugars, which resulted in
different reducing sugar contents among the treatments.
A general decease in reducing sugars was also noted in
the fermentation of carrot and other foods25.
Sensory Evaluation
The sensory score of tempoyak as evaluated by an
18-member panel is given in Table 4. Analysis of
variance of the sensory means a score of different
treatments. There is no significant difference among the
samples as tested at the 0.05 level of significance for
sourness or aroma. However, there was a significant
difference among treatments for general acceptability.
The P. acidilactici UP02 inoculated samples were more
acceptable than those with no-starter treatment.
In general, the results of the sensory evaluation
revealed that the judges were not able to distinguish any
difference among the treatments on sourness and aroma.
This might be attributed to the fact that 8-d fermented
samples were presented in the sensory, wherein, based
on their biochemical analysis, pH, and acidity were not
significantly different among the treatments. Besides,
the predominant component present in all the samples
was basically the same, that is, sulphur compounds.
Even though sensory attributes were not significantly
309
different among the treatments when it came to aroma
and sourness, the results revealed that samples with P.
acidilactici UP02 treatment had higher scores for
sourness than other samples. This might be related to the
higher acceptability of panelists to the samples treated
with P. acidilactici UP02.
Conclusions
Generally, flavour of tempoyak (fermented durian)
was influenced by the addition of P. acidilactici UP02
as a starter. Addition of P. acidilactici UP02 in
tempoyak fermentation decreased the quantity of
sulphur components, increased various non-sulphur
compounds, values of non-volatile acidity, and the
content of organic acids. The flavour components of
tempoyak with P. acidilactici UP02 treatment were
observed to have 48 compounds, although, those with
no-starter treatments had 22 compounds. The patterns
of reducing sugars in both the samples from day-0 to
day-8 of the fermentation period were similar. The
tempoyak prepared by starter with P. acidilactici
UP02 was more acceptable than the no-starter
treatment.
Acknowledgement
The authors thank SEARCA, IFST UPLB and the
University of Lampung for supporting this work.
References
1 Irwandi & Che-Man Y B, Durian leather: Development,
properties and storage stability, J Food Qual, 19 (1996) 439-
489.
2 Ganjar I, Fermentation of the Far East, in Encyclopedia of
food microbiology, edited by R K Robinson, C A Batt and P
D Patell (Academic Press, London) 2000, 722.
3 Battcock M & Ali S A, Fermented fruits and vegetables, a
global perspective, Chapter 5, FAO Agricultural Services
Bull 134 (FAO, Rome) 1998, 2.
4 Yuliana N, Biochemical changes in fermented durian (Durio
zibethinus Murr). Ph D Thesis. University of the Philippines,
Los Banos, Laguna, Philippines, 2004.
5 Yuliana N, Organic acids component of tempoyak
(fermented durian), J Teknologi dan Industri Pangan, 16
(2005) 90-95.
6 Merican Z, Malaysian tempoyak, in Handbook of indigenous
fermented foods, edited by K H Steinkraus, R E Cullen, C S
Pederson, L F Nellis and B K Gavitt (Marcel Dekker, New
York) 1997, 128-130.
7 Halim D, Study of chemical and microbilogical changes in
tempoyak during fermentation (Minithesis, Institut Pertanian
Bogor, Indenesia) 1985.
8 Nurainy F, Chemical and microbiological aspect of
tempoyak fermentation. Abstract of BS Thesis. University of
Gadjah Mada (UGM), Yogyakarta, 1991.
310 INDIAN J BIOTECHNOL, JULY 2009
9 Amin A M, Jaafar Z & Khim N L, Effect of salt on tempoyak
fermentation and sensory evaluation, J Biol Sci, 4 (2004) 650-
653.
10 Leisner J J, Vancanneyy M, Rusul B, Lefebre K, Pot B et al,
Identification of lactic acid bacteria constituting the
predominating microflora in an acid-fermented condiment
(tempoyak) in Malaysia, Int J Food Microbiol, 63 (2001) 149-
157.
11 Issa Z M, Molecular characterization of Lactobacillus
plantarum isolated from Malaysian fermented foods. Abstract
of MS Thesis, University of Putra, Malaysia, 2000.
12 Wirawati C U, Potential of lactic acid bacteria isolated from
tempoyak as probiotic. MS Thesis, Institute of Pertanian
Bogor, Indonesia) 2002.
13 Leisner J J, Vancanneyy M, Lefebvre K, Vandemeulebroecke
K, Hoste B et al, Lactobacillus durianis sp nov, isolated from
an acid-fermented condiment (tempoyak) in Malaysia, Int J
Syst Evol Microbiol, 52 (2002) 927-931.
14 Leal-Sanchez M V, Ruiz-Barba J L, Sanchez A H, Rejano L,
Jimenez-Diaz R et al, Fermentation profile and optimization of
green olive fermentation using Lactobacillus plantarum
LPCO10 as a starter culture, J Food Microbiol, 20 (2003) 421-
430.
15 Wang Y-C, Yu R-C & Chou C-C, Antioxidative activities of
soymik fermented with lactic acid bacteria and bifidobacteria,
J Food Microbiol, 23 (2006) 128-135.
16 Yin L-J, Pan C-L & Jiang S-T, Effect of lactic acid bacterial
fermentation on the characteristics of minced mackerel, J Food
Sci, 67 (2002)786-791.
17 Cogan T M, Flavour production by dairy starter cultures, Appl
Bacteriol, 79 (1995) 49S-64S.
18 Ramos A, Jordan K N, Cogan T M & Santos H, 13 C-NMR
studies of citrate and glucose co-metabolism by Lactococcus
lactis, Appl Environ Microbiol, 60 (1994) 1739-1748.
19 Oude-Elferink S J W H, Krooneman J, Gottschal J C,
Spoelstra S F, Faber F et al, Anaerobic conversion of lactic
acid to acetic acid and 1,2- propanediol by Lactobacillus
buchneri, Appl Environ Microbiol, 67 (2001) 125-132.
20 AOAC. Official methods of analysis of the association of
official analytical chemist, Chapter 28 (Association of Official
Analytical Chemists, Washington DC) 2000, 9.
21 Nelson N, A photometric adaptation of the Somogyi method
for the determination of glucose, J Biol Chem, 153 (1994) 375-
380.
22 Kim H J & Sohn K H, Flavour compounds of dongchimi soup
by different fermentation temperature and salt concentration,
Food Sci Biotechnol, 10 (2001) 236-240.
23 Mabesa L B, Sensory evaluation of foods: Principles and
methods (Institute of Food Science and Technology, UPLB.
Laguna, Philippines) 1986, 43-45.
24 Ray B & Hoover D G, Pediocins, in Bacteriocins of lactic acid
bacteria, edited by D G Hoover and L R Steensen (Academic
Press, San Diego) 1993, 181-183.
25 Steinkraus K H, Cullen R E, Pederson C S & Gavitt B K,
Handbook of indigenous fermented foods (Marcel Dekker,
New York) 1983, 669.
26 Baldry J, Dougan J & Howard G E, Volatile flavouring
constituents of durian, Phytochemistry, 11 (1972) 2081-2084.
27 Moser R, Duvel D & Grevet R, Volatile constituents and fatty
acid composition of lipids in Durio zibethinus, Phytochemistry,
19 (1980) 79-81.
28 Weenen H, Koolhaas W E & Apriyantono A, Sulphur-
containing volatiles of durian fruits (Durio zibethinus Murr.), J
Agric Food Chem, 44 (1996) 3291-3293.
29 Martin F W, Durian and Mangosteen, in Tropical and
subtropical fruits: Composition, properties and uses, edited by
S Nagy and P E Shaw (AVI Publishing Inc, Westport,
Connecticut) 1980, 407-414.
30 Brown M J, Durio, a bibliographic review (APO International
Plant Genetic Resources Institute, New Delhi) 1977, 1-188.