INTERNATIONAL COLLEGE

UNIVERSITI KUALA LUMPUR

COURSE CODE NKB 20102

COURSE TITLE INTRODUCTION TO BIOCHEMICAL

ENGINEERING

SEMESTER JULY 2017

STUDENT NAME ARINA HAZIRAH BINTI ZULKEFLI

STUDENT ID 55220116087

SUBMISSION TITLE FOOD INDUSTRY

- DAIRY PRODUCTION
Introduction

Enzyme are protein (biological molecules) that have catalytic functions

indispensable to maintenance and activity of life .Enzyme are being used in many
production nowadays to make many products. Important of enzyme has been
discovered by the scientist like aiding in digestion and metabolism and it also help to
break large molecules into smaller pieces as it will make our body easily to absorb it.
There are many industries use enzyme in order to produce their product. One of them
are in food industry. For dairy production, most enzyme that are used rennet,
proteases, lactases, esterase and catalase. Every enzyme has its own function to
certain products and it can obtain from by plant, animal, bacteria and microbial.

Objective

 To know the details of specific enzyme for making cheese

 To identify types of coagulants used to make cheese

Rennet

Cheese was manufactured among the earliest applications of exogenous

enzyme back to approximately 6000 B.C by using rennet. The active ingredient of
rennet is enzyme chymosin that is also known as rennin. The usual source of rennet
is the stomach of slaughtered newly-born calves. Technically rennet is also the term
for the lining of a calf's fourth stomach that we called it abomasum. The reason one of
important enzymes in the business of making cheese is come from calves because
calves able to break down the milk. The enzyme occurs naturally in calves stomach
when they drink their mother’s milk.

Economically, industry identify that they need more modal to grow the calves.
So, scientist identified the gene in the calves that produced the enzyme and
expressed it in harmless bacteria called Escherichia coli .Gene for calf chymosin was
one of the first genes for mammalian enzymes that has been expressed into
microorganism. Structure of the gene as well as properties of the recombinant
chymosin were cloned in many different laboratories of gene for calf prochymosin in
Escherichia coli. The enzymes were identical when observed by immunodiffusion in
gels, but a slight difference was observed by enzyme linked immunosorbent assay.
 Chymosin be the most important part of the rennet to coagulate milk. A little
chymosin goes a long way; 1 part enzyme to 15,000 parts milk does the job of
coagulation in 40 minutes at 35°C from the stomachs of young calves. Bovine and
porcine rennet are also used in combination with calf rennet in portion of (50 : 50 or
30 : 70). There are two main types of substitute coagulants. Firstly is coagulating
enzymes from plants and also from microorganisms. The ability to coagulate is best
shown by plant enzymes but after store the cheese in storage, the cheese will be
having a bitter taste. Recently, most widely used Fermentation-Produced Chymosin of
some microorganisms for example is in Table 1.

Table 1 : Recombinant chymosin preparation at approching legal and commercial acceptance

Source of DNA Microorganisms Company (brand name)

Calf abomasum Kluyveromyces lactis Gist Brocades (Maxiren)

Genccor/ Chr. Honsen

Calf abomasum Aspergillus niger
(Chymogen)

Synthetic Escherichia coli Pfizer (Chymax)

Vegetarian

i) Vegetable Rennet

Cardoon thistle, butterwort, artichoke, nettles, safflower, melon, yellow

bedstraw, and fig leaves are the true vegetable rennet that come from plant.
To extract enzyme from these plants, the plants need to soak in water to
extract thickening enzyme similar to chymosin. Most all traditional
Portuguese and Spanish are made from plant rennet. There are some
challenge in using plant rennet. Some of it are it requires a great skill to
produce a consistent cheese using certain vegetable rennet. Beside, this
cheese will frequently slightly bitter flavor when year come forwards. Below
are the figures of plants that are used in produce cheese.
 Figure 1: Artichoke Figure 2: Butterwort

(Cynara scolymus) (pinguicula)

Figure 3: Cardoon thistle Figure 4: Fig leaves

(Cynara cardunculus) (Ficus carica)

Figure 5: Melon Figure 6: Safflower

(Cucumis melo) (Carthamus tinctorius)

 Figure 7: Yellow bedstraw

(Galium verum)

ii) Microbial enzyme

Owing to an increase in demand for cheese production worldwide, the industrial

want to reduced supply of calf rennet and it led to search rennet substitute such as
microbial rennet. (Stanley,1998). There are many microorganisms that are used
extensively for rennet production in cheese manufacture. For instance, Rhizomucor
pusillus, Rhizomucor miehei, Endothia parasitica, Aspergillus oryzae and Irpex lactis.
Cheese ripening is a complex process mediated by biochemical and biophysical
changes. Therefore, microbial enzyme act as accelerator in cheese ripening.

In order to breakdown carbohydrates, lipids, and proteins by partial or gradual, it

will be mediated by several agents like i) residual coagulant, ii) indigenous milk
enzyme especially proteinases, iii) secondary inocula with their enzymes, iv) starter
bacteria and their enzyme, indigenous milk enzymes and v) nonstarter bacteria and
their enzymes. Enzymatic process in this content is related to cheese ripening which
can be augmenting activity of the key enzymes. Within this, there is advantage if
initiating more specific action for flavor development. If this action can be compared
with elevating the temperature, the result will be accelerating undesirable nonspecific
reactions and consequently off flavor development.

For developing specific flavors in cheese, enzyme will be added for instance
addition of enzyme lipase for the development of Blue-type cheese or Parmesan
flavors. To accelerate ripening, exogenous enzyme are used as the formation of flavor
compounds are largely unknown and it is the most empirical process. In Table 2, it
show different microbial enzymes used to accelerate cheese ripening.
Table 2: Microbial enzyme used to accelearte cheese ripening

Enzyme Source
Microbial lipase Aspergillus niger
A.oryzae
Rhizomucor miehei
Lactase Streptococcus lactis
Kluyveromyces sp.
Escherichia coli
Microbial serine proteinases A.niger
Neutral proteinases Bacillus subtilis
A.oryzae

Figure 8 : Action of rennet on the casein micelle

Based on figure 1, it shows the enzyme in rennet cleaves the casein releasing a
large peptide. The surface of the micelle changes from being hydrophilic and
negatively charged to hydrophobic and neutral. As a result, the micelles aggregate to
trap flat globules and microorganisms in developing curd.
Manufacture of cheese
There are 12 steps in processing cheese :
i. Standardize Milk
ii. Pasteurize/Heat Treat Milk
iii. Cool Milk
iv. Inoculate with Starter & Non-Starter Bacteria and Ripen
v. Add Rennet and Form Curd
vi. Cut Curd and Heat
vii. Drain Whey
viii. Texture Curd
ix. Dry Salt or Brine
x. Form Cheese into Blocks
xi. Store and Age
xii. Package

Figure 9: Manufacture of cheese in industry

All the inquiries like optimum time, temperatures and target pH used for cheese will
depend on individual formulations and the intended use of the cheese.

1. Standardize Milk
Drain is institutionalized to improve the protein to fat proportion to deliver
great quality cheddar with a high return.

2. Pasteurization

Slaughters about all microorganism including pathogens that reason

ailments, for example, yeast and coliforms. Standard temperature is at
72-73°C for 15-20 seconds. Microscopic organisms Clostridium tyrobutyricum
can survive purification as spore and can create butyric corrosive and huge
volumes of hydrogen gas which will pulverize the structure of cheddar.
Substance inhibitors, for example, H2O2 or NaNO3 can be utilized as a part of
a few nations.

3. Cool Milk

Milk is cooled after the procedure of sanitization at 90°F (32°C) to enable the
starter microbes to develop.

4. Inoculate with Starter Starter and Non-Starter Bacteria and Ripen

The starter societies and any non-starter assistant microbes are added to the
drain and held at 90°F (32°C) for 30 minutes to age. The maturing step enables
the microscopic organisms to develop and start maturation, which brings down
the pH and builds up the kind of the cheddar.

5. Add Rennet and Form Curd

After the rennet is included, the curd is not irritated for around 30 minutes so
a firm coagulum shapes. Rennet in powder frame is ordinarily 10 times as solid
as fluid rennet.

6. Cut Curd and Warmth

The curd is then cut with cheddar blades into little pieces and warmed to
100°F (38°C) until the point that it achieves pH 6.4. This warming procedure
isolates the whey from curd.
7. Drain whey

Whey is depleted from the vat to shape a tangle.

8. Texture curd

The curd mats are cut into areas and heaped over each other and flipped
occasionally. This progression is called cheddaring. With this progression, it can
remove more whey and it permits the maturation happen until pH 5.1 to 5.5 is
gotten.

9. Dry Salt or Brine

For cheddar, the littler, processed curd pieces are returned in the vat and
salted by sprinkling dry salt on the curd and blending in the salt. In some cheddar
assortments, for example, mozzarella, the curd is shaped into pieces and
afterward the chunks are put in a brackish water (salt water arrangement).

10. Form Cheese into Blocks

The salted curd pieces are set in cheddar loops and squeezed into squares
to shape the cheddar.

11. Store and Age

Cheddar is put away in coolers until the point when the coveted age is come
to. Cheddar can be matured from a while to quite a while.

12. Package

Cheddar were cut and bundled into pieces and might be waxed.
Conclusion

Taking everything into account, rennet can be get for the most part in stomach of
butchered recently conceived calves or for the vegan they can acquire either from
microbial protein or plants. To remove this compound can be utilized as a part of
numerous courses for instance absorb the plants water to extricate thickening catalyst
like chymosin. Cheddar can be put away for quite a while in the event that we keep it
in right way. The following are some of various kind of cheddar in advertising like
cambabert cheese, swiss cheese , gorgonzola cheese and so on.

Figure10: Camembert cheese Figure11: Gorgonzola cheese

Figure12: Grana cheese Figure 13:Stilton cheese

 Figure 14 :Swiss cheese

References:
1. Qureshi Mehar Afroz, Khare Archana Khan, Pervez Ahmed, Uprit S .(2015).
Enzymes Used in Dairy Industries. International Journal of Applied Research
2015, 1(10), 523-527.
http://www.allresearchjournal.com/archives/2015/vol1issue10/PartH/1-9-198.p
df

2. S. Neelakantan, A. K. Mohanty and Jai K. Kaushik. Production and Use of

Microbial Enzymes for Dairy Processing. Retrieved from
http://www.iisc.ernet.in/currsci/jul10/articles22.htm

3. Christopher Bagley. (2016, February 22).Say Cheese : The real GMO picture.
Retrieved from
http://atransgenicworld.blogspot.my/2016/02/say-cheese-real-gmo-picture.ht
ml

4. Jennifer Meier. (2017, May 08). What is Vegetarian Rennet? Retrieved from
https://www.thespruce.com/what-is-vegetarian-rennet-591572

5. Fifth town. (2008, February). The Importance of Knowing Your Rennet.

Retrieved from
http://www.fifthtown.ca/artisan_cheese/editorial/the_importance_of_knowing_
your_rennet/

6. Kate Arding, Elaine Khosvora. (2011,December 02). Rennet’s Role. Retrieved

from https://culturecheesemag.com/cheese-iq/rennets-role