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Registro: 1
Título: The enzyme effect.
Autores: Kilcawley, Kieran
Fuente: Dairy Industries International; Jul2001, Vol. 66 Issue 7, p26, 3p, 2 diagrams
Tipo de documento: Article
Descriptores: *DAIRY products
CHEESE
FLAVORING essences
NAICS/Códigos del NAICS/Industry Codes 424430 Dairy Product (except Dried or Canned) Merchant
sector: Wholesalers
Resumen: Reports that the enzyme-modified cheeses (EMC) provide suitable alternatives to
natural cheese flavours. Food products in which EMC is cost-effective alternative
to natural cheese; Sources of most proteinases used in EMC; What is important
for the development of EMC flavours.
Recuento total de 2198
palabras:
ISSN: 0308-8197
Número de acceso: 5026962
Base de datos: Business Source Premier
Sección: Cheesemaking technology
THE ENZYME EFFECT
Enzyme-modified cheeses provide suitable alternatives to natural cheese flavours, writes
Kieran Kilcawley
An enzyme-modified cheese (EMC) is a concentrated cheese flavour produced enzymatically from a
range of dairy substrates, usually cheese curd of various ages.

The basic technology to develop EMCs is not new -- preliminary cheese type flavours were produced
in the 1960s. In recent years there has been a renewed interest in EMCs, mainly due to the growth in
demand for convenience foods of which cheese flavour is a major component. EMCs are particularly
suitable for use in such products as they can be tailor-made for different applications.

Cost-effective alternative
In general, EMCs are used as a cheese flavour ingredient in processed foods -- in processed and
analogue cheeses, spreads, snack foods, soups, sauces, biscuits and dips -- where they provide a
cost-effective alternative to natural cheese. Whereas natural cheese is consumed for its flavour,
texture, appearance and functional properties, either alone or as an ingredient, EMCs are consumed
exclusively as a flavour ingredient ( 1).

EMCs can be used as the sole source of cheese flavour, to intensify an existing cheese taste or to
impart a specific cheese character to a more bland product. A wide range of flavours is available,
corresponding to most natural cheese varieties, the most important of which are Cheddar, Blue,
Parmesan and Swiss.

Cheese flavour as an ingredient is generally available either as natural cheese, synthetic cheese
(nature identical or artificial) or EMC. Natural cheese has a number of limitations as a flavour
ingredient in foods due to variations in quality (caused by seasonal differences in milk composition
and differences in the extent of ripening), a lack of flavour intensity and high production costs ( 2, 3).

The disadvantages of using synthetic cheese flavours are mainly due to a lack of knowledge
regarding key flavour compounds for many cheese varieties. For example, no specific Cheddar
flavour compound has ever been explained. Also, with the trend towards the use of natural products,
many food manufacturers may not want to include material which could have a negative impact on
sales.

The main advantages of EMCs are low production costs, product consistency, high flavour intensity,
extended shelf life, low storage costs and the diverse range of flavours available. EMCs are also

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perceived as being natural flavours since they use the same flavour pathways that occur in natural
cheese ripening and are produced via minimal processing techniques. Due to their high favour
intensity, which has been reported to be up to 50 times that of natural cheese, they can also be used
in low fat foods as they are typically added at levels of 0.1% weight per weight.

In addition, EMCs added at this level have no adverse effect on the physical characteristics of the
product into which they are added. This is particularly important in frozen food applications, where a
smooth mouthfeel can be maintained instead of the grainy texture often associated with the use of
natural cheese.

Main pathways
Characteristic flavour in natural cheese is derived from three main pathways, proteolysis, lipolysis and
glycolysis, the influence of which differs depending upon the individual cheese variety ( 4).

However, favour development in EMCs is not as defined as in natural cheese since most of the
flavour is derived through proteolysis and lipolysis. The degree of hydrolysis in EMCs is in general
much greater than that in natural cheese ( 5, 6, 7). It is worth pointing out that the relationship
between the flavour of EMC and its corresponding natural cheese variety remains unclear, illustrated
by the fact that a number of Cheddar-type EMC flavours are available commercially.

Extensive hydrolysis of protein and fat in EMC production is achieved through the addition of
exogenous enzymes to the dairy substrate. The choice of enzymes is dependent upon the final
flavour required and a number of commercial enzyme preparations exist for the production of specific
EMC-type flavours. Most commercial enzyme preparations consist of combinations of proteinases,
peptidases and lipases that have been chosen mainly on the basis of their flavour producing
capabilities.

Starter cultures are also used in the production of EMC flavours. However, little detailed information is
available on their nature, since most remain the proprietary knowledge of individual manufacturers. It
can be assumed that many would be attenuated versions of starter cultures used in natural cheese
production or in accelerated cheese ripening.

Proteolysis in EMC manufacture generates high levels of peptides and free amino acids that
contribute an overall background savoury flavour. The extent of proteolysis required varies: some
commercial Cheddar EMCs were found to have up to four times the levels found in mature natural
Cheddar cheese ( 8).

Most proteinases used in EMC manufacture are microbial and are generally derived from two
sources, Bacillus or Aspergillus spp, and therefore tend to be neutral or acidic in nature. A
consequence of extensive proteolysis can be the generation of high levels of hydrophobic peptides
which impart bitterness in the final product. This is usually unacceptable and can be reduced or
eliminated via controlled proteolysis using specific peptidases, or masked through the addition of
mature cheese or monosodium glutamate.

In practice, bitterness is usually prevented through the use of specific peptidases. It is widely known
that the removal of the amino acid proline from a peptide can greatly reduce its bitterness as it alters
its three-dimensional structure. This changes its solubility and increases its susceptibility to hydrolysis
( 9, 10).

Most commercial peptidase preparations are derived from either Aspergillus or Rhizomucor spp or
lactic acid bacteria. It is not always absolutely necessary to eliminate bitterness. In practice, the final
level of bitterness will be a result of a trade-off between the economics of debittering, desired
functionality and how tolerant the final application is to bitterness ( 10).

Lipolysis is an important flavour pathway in EMC production as it generates specific flavour

compounds and precursors of flavour compounds. Lipolysis involves the hydrolysis of milk fat
triglycerides to free fatty acids and their derivatives.

Short chain volatile free fatty acids are associated with cheese aroma intensity due to their low flavour
thresholds and are therefore linked with the perception of cheese flavour. However, above certain
concentrations they have also been associated with rancidity.

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Longer chain free fatty acids are also important in cheese flavour development but have been
associated with soapy off flavours above certain concentrations. The ratio of short to long chain free
fatty acids is important for development of EMC favours and therefore the specificity of the lipase
used is critical in determining the lipolytic flavour profile produced.

It has been estimated that a suitable lipase preparation for EMC production should contain at least
twice as much activity on esters of four carbons than on esters of six carbons or more ( 1, 6). The
extent of lipolysis in EMC can be considerably higher than in natural cheese. However, the extent will
vary dramatically depending upon the cheese flavour being produced.

Most commercial lipases used in EMC manufacture are derived from either animal or microbial
sources. The most significant animal lipases are isolated from bovine and porcine pancreatic tissues
and the pre-gastric tissues of kid goats, lambs and calves ( 11). Microbial lipases are available from a
wide range of sources, especially Aspergillus, Rhizomucor and Penicillium spp. Microbial lipases are
generally cheaper than animal lipases and have the added advantage of being suitable for vegetarian
and Kosher foods.

Flavour in EMCs can be augmented through the use of potentiators or top notes, such as
monosodium glutamate, yeast extracts, salt, starter distillates and organic acids. These compounds
are used to enhance flavour perception and/or to further expand the product portfolio by imparting
specific flavour notes. Monosodium glutamate and yeast extracts are used to enhance the overall
savoury perception of the product and have the added benefit of masking bitterness or rounding off
sharp notes.

Starter distillates are buttery aromas produced by distillation/concentration of lactococcus

fermentations ( 12), and may contain high levels of acetate, butyrate and diacetyl. These organic
acids impart characteristic flavour notes, reduce the overall pH and have bacteriostatic properties.

Specific enzymes
The basis of EMC production is the use of specific enzymes under optimum conditions to produce
concentrated cheese flavours from dairy substrates, namely cheese curd.

The technology was first developed in the 1960s ( 13), when fresh cheese curd was mixed in a dilute
salt solution to an approximate solid content of 40%, to produce a cheese slurry. Enzymes and
preservatives were added and this mixture was incubated at 30 Celsius for four to five days with daily
agitation, after which characteristic cheese flavours were developed. This work demonstrated the
potential for generating a range of intense cheese flavours from cheese substrates rapidly through
optimising conditions for enzyme activity.

The fundamental process remains similar today, except that processing conditions are highly
optimised and production times can be less than 24 hours. EMCs are generally produced in one of
two ways -- by a one-step process (see Figure 1) where hydrolysis of fat and protein occur
simultaneously, or by a component approach (see Figure 2) where different flavour components are
created separately and then blended ( 14). The component approach allows greater flexibility,
enabling the production of a greater diversity of products.

Another method of EMC production is often used to develop Blue cheese flavours and involves
submerged fermentation using Penicillium roqueforti spores. The substrate typically contains a high
percentage of either partially lipolysed milk fat or cream. Fermentation is carried out under conditions
optimal for growth and metabolism of triglycerols to free fatty acids and methyl ketones, which are
characteristic of Blue cheese flavour.

A key factor in the development of an optimal EMC process is emulsification of the substrate, which
maximises the enzyme substrate ratio.

Stable emulsions are obtained by the addition of emulsifying salts (citrates, orthophosphates,
pyrophosphates and polyphosphates), which solubilise the protein by chelating the calcium. The
protein then becomes an effective emulsifier of the fat ( 6).

The choice of emulsifying salts is important as each has characteristics that govern stability. The
substrate is usually around 40 to 60% solids and generally consists of cheese curd (young, mature or
off-cuts), water and salts.

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The protein content tends to vary between 10 and 25%, with a fat content of 20 to 40%. Additional fat
in the form of butterfat or cream may be added to increase production of volatile cheese flavour
compounds.

The substrate is usually heat treated prior to enzyme addition to ensure indigenous enzymes are
inactivated for greater process control. The conditions used during processing depend mainly upon
the enzymes but are generally between 30 and 45 Celsius with a pH of 5 to 7.

An enclosed vessel offers the best conditions for generation of intense flavours as losses of volatile
compounds are minimised. Continuous agitation is necessary to maintain enzyme substrate
interaction at optimum levels.

Prevention of microbial contamination is also important, as conditions are ideal for the growth of
spoilage organisms. Potassium sorbate or nisin are often added to minimise the possibility of
microbial contamination.

Once the desired flavour has been reached, the enzyme reaction is terminated by heat treatment (72
Celsius for 30 minutes) to produce a stable final product with a standardised flavour intensity and
composition.

Market potential
Today's consumers require convenience foods with a greater choice of flavour, diversity and quality,
which further enhances the market potential of key flavour ingredients such as EMCs.

Current trends in EMC production are towards the development of customised flavours for specific
applications. These are not only targeted at introducing cheese flavour to products, but also to
improve the overall perception and harmonisation of flavour.

As more key cheese flavour compounds and production mechanisms are identified, EMC
manufacturers will be able to further enhance their product portfolios.

DIAGRAM: Figure 1: Enzyme modified cheese production using a one-step approach

DIAGRAM: Figure 2: Enzyme modified cheese production using a component approach

References
(1) Moskowitz, G J et al (1987) Journal of Dairy Science 70, 1761-1769

(2) Freund, P R (1995) International Food Ingredients, No 5 p35-37

(3) Missel, D (1996) Food Product Design 6 p51-59

(4) Fox, P F (1993) In Cheese: Chemistry, Physics and Microbiology, 2nd ed, Vol 1 p1-37

(5) Kilcawley, K N et al (1998) International Dairy Journal 8 p1-10

(6) Kilara, A (1985) Process Biochemistry 20 p35-45

(7) Kilara, A (1985) Process Biochemistry 20 p149-157

(8) Kilcawley, K N et al (2000) International Dairy Journal 10 p181-190

(9) Alder-Nissen, J (1993) In Enzymes in Food Processing 2nd ed, Academic Press, London p159-
203

(10) Pawlett, D et al (1996) Industrial Enzymology 2nd ed, Macmillan Press, London p177-186

(11) Birshback, P (1994) International Dairy Federation Bulletin 294, Brussels p7-10

(12) Law, B A (1999) Technology of Cheesemaking 1st ed, Sheffield Academic Press p163-192

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(13) Kristoffersen, T et al (1967) Journal of Dairy Science 50 p292-297

(14) West, S (1996) Industrial Enzymology 2nd ed, Macmillan Press, London, p211-214

~~~~~~~~
By Kieran Kilcawley

Kieran Kilcawley is research officer at Ireland's National Dairy Products Research Centre. Tel: (+353)
254 2245. Fax: (+353) 254 2340. This article is based on a paper given at the Texel International
Dairy Symposium, a conference organised by Rhodia Food UK. Tel: (+44) 161 910 1516. Fax: (+44)
161 910 1527.

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