ACCELERATE

D CHEESE
RIPENING
VENUS BANSAL
ASSISTANT PROFESSOR
WHAT IS
RIPENING?
• The term ripening / curing / maturing / souring of cheese refers to the storage of cheese at a
specific temperature and humidity, during which its physical, chemical and bacteriological
properties are profoundly changed, resulting in the development of characteristic flavor, body
& texture.

• There is no one cheese flavour compound and the flavour of a particular variety stems from the
combination of a wide range of volatile and non-volatile compounds present in the correct
balance and concentration (the `component balance theory' of cheese flavour).
CHANGES DURING
RIPENING?
 HOW TO MEASURE THE
RIPENING?
• The rate of ripening is measured by determination of the “ripening index as follows”:
 WHY ACCELERATED
RIPENING?
• All the chemical changes which occur during ripening are slow and take about 6 months to
years depending upon the variety of cheese.

• Ripening is a slow and consequently, expensive process that is not fully predictable or
controllable.
 WHAT IS ACCELERATED
RIPENING?
• The process of reducing normal ripening period is called as accelerated ripening.

• The approaches that are used to accelerate cheese ripening process include:

• Use of elevated temperature

• Addition of exogenous enzymes

• Use of adjunct cultures

• Attenuation of starter bacteria / starter cultures

• Genetically engineered starter culture

• Use of high pressure treatment

 EFFECT OF ELEVATED TEMPERATURE

• Elevated temperature is the simplest and cheapest method for accelerated ripening of cheese as no

additional costs are involved and moreover reduces refrigeration costs.

• The temperature used for ripening of most of the variety of cheeses are less than 10º C

• Ripening temperature of some of the varieties of cheese are: Emmental, 22-24º C, Mold and smear-

ripened cheeses- 12-15º C, Dutch varieties- 12-14º C, Cheddar cheese- 6-8º C

• The above temperatures are maximum in the profiles and are usually maintained for 4-6 weeks to induce

the growth of a desired secondary micro-flora. Thereafter, the cheese is transferred to much lower

temperature of less than 10º C

• Storing the cheese at a higher temperature than normal temperature; it will ripen more quickly; the

microorganisms in and on it will grow faster, and enzyme catalyzed reactions will accelerate
 cont …
• Keeping the cheese above 20-25º C causes texture to be too soft and the cheese deforms readily.

• It also causes exudation of fat and excessive loss of moisture.

• Thus the scope for accelerated ripening by elevated temperature is quite limited except for
cheddar cheese which is ripened at low temperature of 6-8º C.

• The ripening time of cheddar cheese can be reduced by 50% by increasing the ripening
temperature from 6 to 13º C without adverse effects.
 ADDITION OF EXOGENOUS
ENZYMES
• Cheese ripening is essentially an enzymatic process and hence by augmenting the activity of
key enzymes; accelerated cheese ripening is possible.
• To produce a balance flavor, a mixture of enzymes in proper ratio has to be added.
• The addition of combinations of various fungal proteases and lipases to cheddar cheese has
been reported to reduce ripening time by 50%.
• A lipase / proteinase preparation derived from Aspergillus oryzae released C6-C10 fatty acids to
produce typical Cheddar cheese flavor.
 CAN RENNET BE USED FOR
ACCELERATED CHEESE RIPENING?
• The principal enzyme present in the rennet is chymosin and is responsible for the primary proteolysis in
most of the cheese varieties.
• Therefore it is expected that ripening could be accelerated by increasing the level of rennet in the cheese
curd.
• However, increasing the rennet in cheese curd does not accelerated ripening and in fact causes bitterness.
• Because chymosin produces only relatively large oligopetides which lacks a typical cheese-like flavor
and may be bitter.
• Chymosin has very little activity on β casein, because principal chymosin-susceptible bond in β casein,
Leu192-Tyr193, is in the hydrophobic C-terminal region of the molecule which appears to interact
hydrophobically in cheese, rendering this bond inaccessible.
• However, use of Cryphonectria parasitica proteinase preferentially hydrolyses β casein in cheese without
causing flavor defects (possibly because its preferred cleavage sites are in the hydrophilic N-terminal
region.
 USE OF PROTEINASES FOR
ACCELERATED CHEESE RIPENING
• The use of proteinases enzymes to cheese milk accelerates the ripening of cheese but the
principle problem associated with this approach poses several problems like:
 Proteinases are usually water soluble, and when added to cheese milk, most of the enzyme
is lost in whey, which increases cost.
 The prohibition of use of exogenous enzymes in many countries.
 The uniform distribution of enzyme in the curd.
 Enzyme-contaminated whey must be heat treated if the whey has to be utilized. Moreover,
the choice of enzyme is limited to those that are inactivated at temperatures below those that
cause thermal denaturation of whey proteins.
 The addition of enzyme in milk decreases the yield of the cheese due to hydrolyzation of β
casein which lost in whey during pressing
 USE OF PLASMIN FOR
ACCELERATED CHEESE RIPENING
• The use of proteinase enzyme like plasmin to cheese milk accelerates the ripening of cheese
without any off-flavor development.

• Plasmin is associated with the casein micelles in milk, which can bind at least 10 times the
amount of plasmin normally present, and is totally and uniformly incorporated into cheese
curd, thus overcoming one of the major problem encountered with use of exogenous
proteinases to accelerate cheese ripening.
 EXOGENOUS LIPASES &
LACTASE
• Exogenous lipases
 Rennet paste or crude preparations of pre-gastric esterase (PGE) are normally used in many
cheese.
 Lipases from Penicllium requeforti and Penicillium candidum are also used for accelerated
ripening of cheese
• Lactase (β-Galctosidase)
 It hydrolyses lactose to glucose and galactose, results in stimulation of LAB and thus
shortens the lag period.
 Lactase from Kluyveromyces lactis available as Maxilact (Brand name) contains a
proteinase, which is responsible for incrased levels of peptides and free amino acids.
 USE OF ADJUNCT CULTURES
• Adjunct cultures are specifically selected strains, which are intentionally added to accelerate
ripening of cheese.

• The principal adjuncts used in the accelerated ripening of cheese are mesophilic Lactobacilli
and thermophilic Lactobacilli.

• Inoculation with mesophilic Lactobacillus adjuncts enhanced flavor and accelerated proteolysis
at the level of small peptides and amino acids.

• In contrast to mesophilic Lactobacilli, thermophilic Lactobacilli die rapidly in cheese, lyse and
release their intracellular enzymes. Consequently, cheese s made with thermophilic
Lactobacillus spp. as starters contain high concentrations of amino acids
• Augmentation of starter culture with L. casei had definite and positive influence the flavor,
body & texture of buffalo milk cheddar cheese.

• Addition of different Lactobacillus spp. (L. casei and L. plantarum) to cheddar cheese milk to
a level of 105-106 cfu / ml increased the level of free amino acids to attain highest flavor scores.
 ATTENUATION OF STARTER
CULTURE
• Starter cultures are an obvious choice as ripening agents to control and accelerate the ripening
of cheese. But increasing the rate of culture results in more / faster acid production which
results in a different type of cheese.

• Attenuation of Starter cultures i.e. heat and freeze shock treatments, exposure to lysozyme
under conditions which do not lyse the starter and selection of natural variants which can not
ferment lactose can be used for accelerated ripening of cheese.

• The acid producing ability of lactic acid bacteria can be markedly reduced by a sub-lethal heat
treatment while only slightly reducing enzyme activities. Heating at 59 and 69º C for 15
seconds is optimal for mixed mesophilic and thermophilic Lactobacilli cultures respectively.
• Addition of lysozyme sensitized cells to cheese milk at a level of equivalent of 10 10 cells / g of
cheese indicated that intracellular dipeptidases were released and as a result, concentration of
free amino acids significantly increased.

• However, there was no effect on the rate of flavor development.

• Economically, the use of lysozyme treated may not be viable owing to the cost of the enzyme.
 GENETICALLY ENGINEERED
STARTERS
• GM LAB which can be applied to cheese-ripening and cheese flavour technology have been
changed with respect to either their sugar metabolism, peptidase production capacity or their
rate of lysis in the matrix of freshly-made cheese.

Genetic modification Change from wild type Ripening / flavor effect

Lactose metabolism diverted by Increased production of Enhanced buttery flavor, sharper

selective elimination and enhancement diacetyl and acetic acid background taste
of key metabolic pathway enzymes
Changed proteinase and peptidase Specific increase in Reduction in bitterness, enhancement
balance, selectively enhanced peptidase concentrations of of overall taste intensity, increase in
production aromagenic amino acid sweet, sulphurous, floral or nutty
concentrations in cheese flavor notes
Introduction of cell lysine genes under Controlled instantaneous Faster release of intracellular flavor-
control of externally triggered lysis in cheese matrix producing enzymes
promoters
 USE OF HIGH PRESSURE
TREATMENT
• It is one of the technique used for accelerated cheese ripening, however, additional research is required to
define the operating conditions of HP treatment to provide positive effects in cheese making and
ripening.

• The application of HPT to cheese results in an increase in moisture content and pH and cause changes to
the cheese matrix and lysis of the cells, which contribute to ripening.

• An increase in pressure tends to results in a decrease in volume, which enhances chemical reactions,
phase transitions and changes in molecular configurations.

• The pressure used for accelerated ripening of cheese ranges from 100-1000 Mpa for a very short period.

• Use of HHP treatments increase pH (0.1 to 0.7 units) and modify water distribution, leading to enhanced
condition for enzymatic activity.