Rennet production

S.Shanmuga Priya
BTF-07-023
 Rennet
•
• Rennet is in use for cheese making at
least since 2000 B.C.
• An enzyme extracted from gastric
mucous membrane of bovines
• The production of countless different
types of cheese use proteolytic
enzyme, rennin the one most
widely used
 Cheese
§Cottage cheese §Limburger
§Roquefort §Traditional brick cheese
§Stilton §Cheddar
§Blue cheese §Colby
§Gorgonzola §Sweet Brick
§Camembert §Muenster
§Brie §Havarti
§Swiss §Mozzarella
§Baby Swiss §Provolone
§Gouda §Parmesan
§Edam §Romano
 Alternatives source of
rennet
• Global increase in cheese production
• Shortage of calf rennet
• Demand for meat made more calves
reared to maturity
• Let to high cost of natural calf rennet
and find alternate proteolytic
enzymes that would duplicate the
action of calf rennet in cheese vat.
•
 Microbial rennet
• Some molds such as Rhizomucor
miehei, Mucor miehei, Mucor
pusillis and Endothia parasiticaare
able to produce proteolytic
enzymes
• The flavor and taste of cheeses
produced with microbial rennet's
tend towards some bitterness,
especially after longer maturation
periods.
 Genetically Engineered
Rennet
• Calf genes to modify some bacteria,
fungi or yeasts to make them
produce chymosin
• Recombinant technique -
Recombinant chymosin
• the first artificially produced enzyme
registered and allowed by the U.S.
Food and Drug Administration
•
 Conti….
• 60% of U.S. hard
cheese was made
with genetically
engineered
chymosin
• it has up to 80% of
the global market
share for rennet.
• the most widely used genetically
engineered rennet is produced by the
fungus Aspergillus niger
• Other micro organisms are Escherichia
coli, Kluyveromyces lactis
• Escherichia coli
– not regarded as GRAS (Generally
Regarded As Safe)
– recombinant proteins are frequently
synthesized as intracellular inclusion
bodies process cost increases
• Cheese production with chymosin is
similar to production with natural calf
rennet
• Preparation of calf rennet has two
forms of chymosin A in proportion
40% & B 60 %
• Genetic rennet contains only one of the
known main chymosin types, either
type A or type B
• Other do not exist
 Conti…..
• genetically engineered chymosin and
natural pepsin is mixed to imitate
the complexity of natural rennet
and to get the same results in
coagulation and in development of
flavor and taste
• suitable for vegetarians if there was
no animal based alimentation used
during the production in the
fermenter
 Aspergillus niger
• Aspergillus niger is a fungus and one of
the most common species of the
genus Aspergillus
• a common contaminant of food
• A. niger is cultured for the industrial
production of many substances.
• Various strains of A. niger are used in
the industrial preparation of citric acid
(E330) and gluconoic acid (E574)
• A. niger fermentation is "generally
regarded as safe“ by the FDA.
 Fermentation

The production organism is grown in several stages to build

up the inoculums for large-scale production
After inoculation the cells grown aerobically under the proper
conditions of pH, temperature, nutrient composition
After chymosin reaches a desired level in the fermentation broth, the
fermentation is stopped and the fungal cells are inactivated and
separated from the liquid.
• Chymosin recovered from the broth -
two methods:
• the broth is filtered, followed by
chromatographic purification and
concentration of chymosin
• the chromatographic step is preceded
by extraction of chymosin from the
fermentation broth
• Chymosin eluted from the
chromatographic column is
formulated to the commercial
Rennet producing industries
• Gist brocades – maxiren – K.lactis
• Genecor / Chr. Hansen – chymogen –
A.niger
• Pfizer – chymax- E.coli
• Iranian industrial enzymes company
– rennilase