Industrially important fungal

enzyme: PROTEINASE
Rajani prabhu
MI-1414
Msc part 1
Microbiology.

 Recent years have witnessed a phenomenal increase in the use of

enzymes as industrial catalysts.

Proteases (EC 3:4, 11-19, 20-24, 99) (synonymous as peptidase or proteinase) constitute a very large and complex group of enzymes, widely
utilized in most of industries.
They differ in properties such as substrate specificity, active site and
catalytic mechanism, pH and temperature optima, and stability
profiles.
The main sources of the enzymes were from animals (e.g. calf
stomach), plants (e.g. pineapple, fig, and papaya), microbes (e.g.
Bacillus spp., Pseudomonas spp.) etc.
But the production of enzymes from plant and animal sources is
limited due to climatic reasons and ethical issues, respectively.
Microbial sources have occupied an invincible domain in the
production of all the threeacidic, neutral, and alkalinemajor types
of proteases.

classification
On the basis of their site of action on protein

substrates, proteases are broadly classified as endoor exo-enzymes.

Proteases are also classified into different clans and
families depending on their amino acid sequences and
evolutionary relationships.
Based on the pH optima, they are referred to as
acidic, neutral, or alkaline proteases.
They are further categorized as serine proteases,
aspartic proteases,glutamic acid proteses,threonine
proteases cysteine proteases or met- allo proteases
depending on their catalytic mechanism (Table 1).

Table 1. General classification of proteases with their

enzyme commission (EC) code, coupled with specific
mechanism of action of each subgroup.
Protease EC code Mechanism

Exopeptidases 3, 4, 11-19

cleave the peptide bond proximal to the amino or carboxy termini of

the substrate

Aminopeptidases 3, 4, 11 Those acting at a free N-terminus liberate a single amino acid residue

Dipeptidases 3, 4, 13 Exopeptidases specific for dipeptides

Dipeptidyl peptidase
3, 4, 14 Release of an N-terminal dipeptide from a polypeptide

Tripeptidyl peptidase
3, 4, 14 Release of an N-terminal tripeptide from a polypeptide

Peptidyldipeptidase
3, 4, 15 Release of free C-terminus liberate a dipeptide

Carboxypeptidase
3, 4, 16-18 Release of a single residue C-terminal from a polypeptide

Serine type protease

3, 4, 16 Carboxypeptidase have an active centre serine involved in
the catalytic process

Metalloprotease
3, 4, 17 Carboxypeptidase use a metal ion in the catalytic mechanism

Cysteine type protease 3, 4, 18 Carboxypeptidase have a cysteine in the active centre

Omega peptidases 3, 4, 19 Remove terminal residues that are linked by isopeptide bonds

Endopeptidases 3, 4, 21-24 Cleave internal bonds in polypeptide chains

Serine protease 3, 4, 21 Endopeptidases have an active centre serine involved in the catalytic
process

Cysteine protease 3, 4, 22 Possesses a cysteine in the active centre

Aspartic protease 3, 4, 23 An aspartic acid residue for their catalytic activity

Metalloprotease 3, 4, 24 Use a metal ion (often, but not always, Zn2+) in the catalytic mechanism

Endopeptidases of
3, 4, 99 Acting on peptide bonds
unknown catalytic mechanism

sources
Animal Proteases: The most familiar proteases of

animal origin are pancreatic trypsin, chymotrypsin,

pepsin and rennin.
Pepsin is an acidic protease that is found in the
stomach of almost all vertebrates.
Plant Proteases: Papain, bromelin, keratinases, and
ficin are some of the well-known proteases of plant
origin.
Microbial proteases: microbial community is
preferred over the others for the large scale
production of proteases due to their fast growth and
simplicity of life for the generation of new
recombinant enzymes with desired properties

Fungal proteases is advantageous

Fungal proteases: Fungal proteases

magnetized the interest of researches due to

high diversity, broad substrate specificity,
and stability under extreme conditions
it offers an advantage of separation of
mycelium by simple filtration.
Fungal proteases can conveniently be
produced in solid-state fermentation process.
Fungal proteases are also used in for
modifying food proteins and various other
industrial applications.

Sources of commercially significant

proteases
To meet commercial demands, large scale

production of these enzymes became necessary

and hence several sources were explored
including fungi.
The fungi possess diverse habitat, grow
ubiquitously and produce various
proteins/enzymes their survival. Among these
enzyme proteases.
Based on habitat fungi are classified as
psychrophilic, mesophilic and thermophilic
emerged as potential sources of commercial
enzymes including proteases.

Psychrophilic fungi: capable in producing

cold tolerant protease.

Several deep sea fungi are capable in
producing psychrophilic protease including
Aspergillus terreus, Beauveria brigniartii
and Acremonium butyri.
used in food processing,fruit and milk
processing industries.

Thermophilic fungi:capable in producing

commercial protease that act in higher
temperature such as Canariomyces
thermophila, Thermomyces ibadanensis,
Talaromyces thermophiles, Myriococcum
and Dactylomyces thermophiles.
These proteases not only offer catalytic
activity in
higher temperature but also possess longer
shelf life
and provide ease in storage.
It has applications in leather ,detergent,

Mesophililic fungi :The mesophilic fungal

strains are also competent in production of

large scale of commercial protease
enzymes. The major class of mesophilic
fungi is Aspergillus which contribute more
than 25% of protease produced from fungal
source.
Have applications in
paper,pulp,leather,textile,detergent,agricult
ure,animal feed and food processing
indutries.

Protease in food processing

Food processing utilizes several enzymes for

production of quality and nutritional food

products.
Proteases have important roles in baking,
brewing and in the production of various
oriental foods such as soy sauce, miso, meat
tenderization and cheese manufacture.
Further, processing of fruits and storage of
their
juices for long shelf life need enzyme treatment
and primarily protease are employed.
Preparation of milk products and candies
need complete processing catalysed by series
of enzyme including protease.

Beverage industry both soft and hard drink need a

complete processing of substrate for elegant flavour

and shelf life of product.
Processing of tea, coffee and coca needed a
precise oxidation of raw material
(ripen seeds, leaves and berry) to produce complete
products and enzyme such as protease play crucial
role.
Ethanol is product of sugar fermentation
carried out by series of biochemical
reactions catalysed by group of enzyme
including different type of proteases

Protease in textile industry

Another most significant area of commercial application of protease is textile

industry where a final elegant texture is provided by enzyme treatment.

Silk is processed by thermostable protease to remove gum and other
impurities lies of core protein fibre on silk.
Silk degumming a growing industry, enormously utilize protease and results
in high quality silk.
Synthetic fabric also subjected to protease treatment for elegant and
smooth finish.
Fungal proteases are key enzyme in fabric industry since decade and
growing tremendously.
Indian sericulture has grown double in last one decade and consumption of
protease also enhances in several folds.
Use of such method not only offer good quality fabric but also impart
mechanical strength to fibre.
Use of protease minimizes chemical detergent associated
with large scale of
environment pollution.

Protease in leather making

Leather is prime commodity for Indian in earning foreign currency and India is

the second largest exporter of processes and unprocessed leather on world

map after china
Leather processing involves several steps such as soaking, dehairing, bating,
and tanning.
Old and conventional methods used are less efficient and utilized massive
amount of chemical that leads lead to environmental pollution.
Several studies have suggested that the enzymatic processing of leather is
environment friendly and yield better quality leather.
Leather making which primarily carried out at higher temperature involves
several thermostable proteases
. Thermophilic fungi have emerged as potential sources for leather processing
Further, cleaning of hair and their biological digestion had yielded in several
vital amino acids used as dietary supplement
Several studies have been carried out to reuse of
these potential enzyme after immobilization for
leather making

Proteases as detergents
Biological detergent had great attention in current scenario primarily protease.
Both domestic and industrial detergents are having major constituents from

protease.
Proteases are one of the standard ingredients of all kinds of detergents
ranging from those used for household laundering to reagents used for
cleaning contact lenses or dentures
The biological detergent are primarily used in cleaning of large boiler in
industries, hospital,poultry industry.
Discovery of thermostable protease further enhanced efficiency of biological
detergent.
In modern days both domestic and industrial detergent are utilizing protease
in higher extent.
In order to meet commercial demand fungal enzymes
have become key source for detergents.
Fungal alkaline proteases are advantageous due
to the ease of downstream processing to prepare
a microbe-free enzyme.

Therapeutic applications
The medicinal application of protease for

diagnostic and therapeutic is widely accepted and

several enzymes are in use since many years.
Proteases are primarily associated with
development of anticancer-caspase, antiinflammatory-Serratiopepetidaseses,
antimicrobial-cytotoxic nature,clot dissolving
agents-thrombolytic.
Oral administration of protease from Aspergillus
oryzae (Luizym and Nortase) has been used
as a digestive aid to correct certain lytic
enzyme deficiency syndromes.

REFERANCES..
Izabel Soares, Zacarias Tavora, Rodrigo Patera Barcelos

and Suzymeire Baroni (2012). Microorganism- Produced

Enzymes in the Food Industry, Scientific, Health and
Social Aspects of the Food Industry, Dr. Benjamin Valdez
(Ed.), ISBN: 978-953-307-916-5, InTech.
J. Srilakshmi, J. Madhavi, Lavanya S, Ammani K(2014)
Commercial Potential of Fungal Protease: Past, Present
and Future Prospects. J.Pharmaceutical , Chemical and
Biological Sciences, 2(4): 218-234.
Rao Mala et.al,(1998) Molecular and Biotechnological
Aspects of Microbial Proteases. J.microbiol mol biol rev.
62(3): 597635.
V. N. Jisha et al. (2013) versitality of microbial proteases.J.
Advances in Enzyme Research1(3):39-51

Thank you.