What is the role of protease?

One of the main functions of proteases is to process proteins. Proteins in the body are difficult to digest and do not contain enzymes. Other types of proteases are involved in regulating the activities of blood clotting cells. These enzymes are also called proteolytic enzymes.

Proteins are held together by peptide bonds. Long-chain amino acids, small pieces of protein are called peptides, large pieces of protein are called peptides, and enzymes that break down peptides are called proteases. Proteases are types of proteins that accelerate degradation. γpeptidase cleaves terminal amino acids to break down peptide bonds and release amino acids produced due to residual protein.

What is protease?

Proteasomes are ubiquitous in eukaryotes and archaea, and a giant protein complex that also exists in some prokaryotes. In eukaryotes, the proteasome is located in the nucleus and cytoplasm. The main role of the proteasome is to degrade proteins that are not needed or damaged by the cell. This role is achieved by chemical reactions that break peptide bonds. Enzymes that can perform this role are called proteases.

The proteasome is the main mechanism used by cells to regulate specific proteins and remove misfolded proteins. After proteasome degradation, the protein is cleaved into peptides about 7-8 amino acids long; these peptides can be further degraded into single amino acid molecules, and then used to synthesize new proteins. The protein that needs to be degraded is first labeled (ie attached) by a small protein called ubiquitin. This labeling reaction is catalyzed by ubiquitin ligase. Once a protein is labeled with an ubiquitin molecule, it will trigger other ligases to add more ubiquitin molecules; this forms a “polyubiquitin chain” that can bind to the proteasome, thereby bringing the proteasome here. A labeled protein begins its degradation process.

From the structural point of view, the proteasome is a barrel-shaped complex, including a “core” composed of four stacked rings. The core is hollow to form a cavity. Among them, each ring is composed of seven protein molecules. The middle two loops each consist of seven β subunits and contain six active sites for proteases. These sites are located on the inner surface of the loop, so the protein must enter the “cavity” of the proteasome to be degraded.

The two outer rings each contain seven α subunits, which can function as a “gate” and are the only way for proteins to enter the “cavity”. These alpha subunits, or “gates”, are controlled by the “cap”-like structures (ie, regulatory particles) attached to them; the regulatory particles can recognize the polyubiquitin chain tag attached to the protein and initiate the degradation process. The entire system including ubiquitination and proteasome degradation is called the “ubiquitin-proteasome system”. The proteasome degradation pathway is essential for many cell processes, including the cell cycle, regulation of gene expression, and oxidative stress.

The winning themes of the Nobel Prize in Chemistry in 2004 were the importance of protein enzymolysis in cells and the role of ubiquitin in the enzymatic pathway. The three winners are Aaron Chehanovo, Avram Hershko and Owen Rose.

What is biological protease?

Biological protease is an abbreviation of enzyme that hydrolyzes protein peptide bond. This substance is generally widely distributed in plant stems and leaves, animal organs, microorganisms and fruits. The protease can accelerate the digestion of food after entering the human body, and has a relatively good help for indigestion that occurs in the human body. Protease also has a better recovery effect for some other types of diseases.

What is the role of pepsin?

Pepsin is a digestive protease whose main function is to break down the protein in food into small peptide fragments. Pepsin is mainly used to treat indigestion caused by eating too much protein food, or recovery period after serious illness, impaired digestion and chronic atrophic gastritis, as well as impaired digestive function of gastric cancer, and protease deficiency caused by pernicious anemia.

As biocatalysts, enzymes and general catalysts are the same in many respects. For example, the amount is small but the catalytic efficiency is high. Like general catalysts, enzymes can only change the speed of chemical reactions, not the equilibrium point of chemical reactions. The enzyme itself does not change before and after the reaction, so the relatively low content of the enzyme in the cell can catalyze the change of a large number of substrates in a short time, reflecting the high efficiency of enzyme catalysis. Enzymes can reduce the activation energy of the reaction, but do not change the free energy change (³G) during the reaction, thus speeding up the reaction and shortening the time for the reaction to reach equilibrium, but it does not change the equilibrium constant.

Compared with general catalysts, the catalysis of enzymes shows unique characteristics.

1. High efficiency of enzyme catalysis

The catalytic activity of enzymes is much higher than that of chemical catalysts. For example, catalase (containing Fe2+) and inorganic iron ions both catalyze the decomposition reaction of hydrogen peroxide as follows. 1 mol of catalase can catalyze the decomposition of 5×106 mol of H2O2 in 1 minute. Under the same conditions, 1 mol of the chemical catalyst Fe2+ can only catalyze the decomposition of 6×10-4 mol of H2O2. Compared with the two, the catalytic efficiency of catalase is about 1010 times that of Fe2+.

The level of enzyme catalytic efficiency can be expressed by the concept of turnover number. The conversion number refers to the number of molecules per enzyme molecule that can convert the substrate per minute when the substrate concentration is large enough, that is, the number of molecules that catalyze the chemical change of the substrate. According to the data introduced above, the conversion number of catalase can be calculated as 5×106. The conversion number of most enzymes is around 1,000, and the largest can reach over 106.

(2) High specificity of enzyme catalysis

An enzyme can only act on a certain type or a specific substance. This is the specificity of enzyme action. For example, glycosidic bonds, ester bonds, peptide bonds, etc. can be hydrolyzed by acid-base catalysis, but the enzymes that hydrolyze these chemical bonds are different. They are the corresponding glycosidases, esterases, and peptidases, that is, they are individually specific. It can be hydrolyzed by natural enzymes.

(3) Mild reaction conditions catalyzed by enzymes

Enzymatic reactions generally require mild conditions such as normal temperature, normal pressure, and neutral pH. Because enzymes are proteins, they are prone to lose their activity in environments such as high temperature, strong acid, and alkali. Since enzymes are more sensitive to changes in the external environment and are easily denatured and inactivated, the reaction conditions must be strictly controlled during application.

(4) Adjustability of enzyme activity

Compared with chemical catalysts, another feature of enzyme catalysis is that its catalytic activity can be automatically regulated. Although there are many kinds of chemical reactions in organisms, they are very coordinated and orderly. Changes in substrate concentration, product concentration and environmental conditions may affect enzyme catalytic activity, thereby controlling the coordinated and orderly progress of biochemical reactions. Any disorder and imbalance of the biochemical reaction will inevitably cause the organism to produce disease or even death in severe cases. In order to adapt to changes in the environment and maintain normal life activities, organisms have formed a system that automatically regulates enzyme activities during the long evolutionary process. There are many ways to regulate enzymes, including inhibitor regulation, feedback regulation, covalent modification regulation, zymogen activation, and hormone control.

(5) Enzyme catalytic activity is related to coenzyme, prosthetic group and metal ion

Some enzymes are complex proteins, and the small molecules of coenzymes, cofactor and metal ions are closely related to the catalytic activity of the enzyme. If they are removed, the enzyme loses its activity.

Today, the world is changing at a very fast pace. Innovation & Technology is key. Biology stream is also now witnessing a new kind of improved technology & innovation. Globalization of improved products, services & technology has created a substantial rise in the Bio-economy of the country.

Nowadays, Biotech and Healthcare based startups are enjoying full support from the Government and private sectors, as both these sectors are capable of providing innovative solutions to the global health issues.

Bio-entrepreneurship is now, also considered as a good career opportunity because of improved Incubation support from the government. Biotechnology Incubation is different from the other incubation as biotechnology-based startups require a large space for wet laboratory along with excellent instrumentation facilities to conduct the experiments.

Biotechnology Industry Research Assistance Council (BIRAC), is a not-for-profit state-owned enterprise under the Department of Biotechnology (DBT), Government of India to strategically empower emerging biotech companies.

BIRAC has launched a special scheme, BioNEST (Bioincubators Nurturing Entrepreneurship for Scaling Technologies) to provide Incubation support to the Biotech / Life science-based startups. Incubation support is a must for any Bio-entrepreneur as they need laboratory infrastructure, mentoring support and high tech instrumentation facility to convert their ideas into a prototype. Bio-entrepreneurs are generally young innovators who do not have sufficient funding in the initial days to set up a high tech laboratory or instrumentation facility. Thus, this whole kind of support is provided by special and uniquely designed technology incubator, which is known as Bio-incubator. BioNEST scheme has given the liberty to the leading research institution to develop such infrastructure, which can convert research & innovative ideas into viable commercial entities.

BioNEST scheme is launched by BIRAC with a vision of fostering the biotech innovation ecosystem in the country. BioNEST scheme provides support to establish Bio-Incubators either as a standalone entity or as a part of the academia. Through BioNEST, BIRAC has supported about 50 bio-incubators.

BioNEST scheme came as a boon for high-risk areas related to healthcare, where just a bench space is not enough and innovator need all kind of support. For Biotech startups, the innovator needs good instrumentation facility and technical support. In addition, they also need mentoring from expert scientists and business mentors to succeed.

Top Bio-Incubators are located in Biotech Science Clusters (BSCs), which provide unique Bio-Incubation support to Biotech startups. Some Bio-incubators also provide manufacturing facilities to support the growth of very young Biotech startups. Bio-Incubator under BioNEST work as a support system and provide necessary infrastructure and incubation facilities to the selected startups related to Biotechnology and healthcare.

BSC BioNEST Bio-Incubator (BBB) is one such Bio-Incubator located in NCR Biotech Science Cluster.

BBB supports young startups by providing globally competitive incubation services. Emerging companies have professional and technical assistance, shared space, mentors’ supervision and other additional services to get on a fast track to success. BBB stands firmly with its mission to stimulate the establishment and growth of biotech startup companies and hence enhances the economic wealth of the region by promoting scientific enterprise development.

BBB provides excellent incubation facilities & infrastructure spread across 35000 sq.ft. covered area. Innovators have a variety of options and facilities under a single roof. It includes Office Space, Culture Facility, Lab Space, Professional Business Suites, Meeting rooms, Seminar rooms, Video conferencing facility as well as Instrumentation Facility.

BBB also provides shared wet lab benches for young startups as well as independent lab cubicles for an innovator/ startups that need bigger lab spaces. The best thing about BBB is its unique infrastructure which is dedicated towards healthcare based startup and flexibility in options to choose from shared or independent wet labs with a seating capacity of 3, 4, 8 and 12 respectively which outline other Bio-Incubators. The facility is available to the biotech startups at a very affordable cost.

BBB provides all kind of critical services to the startups which are required to move them on the fast track to success. The infrastructure is designed in such a way that it meets future Biotech innovation challenge and provide globally competitive incubation facility and support.

Incubation support of BBB is first of its kind incubation in Delhi-NCR focusing on innovations in Biotechnology related areas.

Various biochemical reactions continue to occur in organisms, which require corresponding enzymes to catalyze, so there are many types of enzymes. According to statistics from the enzyme website BRENDA, the number of enzymes officially included has reached 7,984.

To study and apply these enzymes with different properties and functions, a systematic and effective classification method is necessary. Although the molecular composition and cellular location of enzymes can be used as the basis for classification, classification according to the type of reaction catalyzed is the most reasonable method for functional research.

In 1961, the International Union of Biochemistry and Molecular Biology (IUBMB) classified all enzymes into six categories according to the types of reactions they catalyzed. In August 2018, the classification of translocation enzymes was added, so there are now seven major enzymes, namely: oxidoreductase (EC 1), Transferases (EC 2), hydrolase (EC 3), lyase (EC 4), isomerase (EC 5), ligase (EC 6) and translocator (EC 7). Among them, EC stands for Enzyme Commission.

Among the seven enzyme classes, each class is divided into several subclasses. The sub-categories differ according to the characteristics of various reactions. For example, oxidoreductase is based on the type of electron donor, and transfer enzyme is based on the type of transferred group, and so on.

Oxidoreductase catalyzes oxidation-reduction reactions, that is, electron transfer between molecules. It is isomerase that catalyzes intramolecular redox reactions. According to the principle of systematic classification, oxidoreductases are divided into 24 subtypes according to the type of electron donor (substrate), and the sub-subtypes are divided according to the electron acceptor.

Traditionally, oxidoreductases are often divided into 4 subtypes: dehydrogenase (receptor is a reducing coenzyme), oxidase (receptor is molecular oxygen), peroxidase (receptor is hydrogen peroxide), and oxygenase (catalyze the incorporation of oxygen atoms into organic molecules). This is also the naming convention for common names of oxidoreductases.

Oxidoreductases is the most abundant enzyme, currently there are 2375 kinds. Its oxidation, productivity, detoxification and other functions are extremely important to organisms, and its application in production is second only to hydrolytic enzymes. Such enzymes usually require cofactors, which can be determined according to the photoelectric properties of the cofactors during the reaction.

Transferase catalyzes the transfer reaction of functional groups, such as various aminotransferases and kinases catalyze the transfer of amino and phosphate groups, respectively. Coenzymes are often needed for transfer enzymes, but the reaction is not easy to determine. According to the nature of the transferred group, it is divided into 10 subclasses, the important ones are:

One-carbon transferase (EC 2.1): transfer one carbon unit, such as methyltransferase related to nucleic acid and protein methylation. Carboxyltransferases belong to this subclass, such as methylmalonyl-CoA carboxytransferase (EC 2.1.3.1). However, carboxylase that consumes ATP is a ligase, such as pyruvate carboxylase (EC 6.4.1.1).

Glycosyltransferase (EC 2.4): closely related to carbohydrate metabolism, such as glycogen synthase (2.4.1.11) and glycogen phosphorylase (2.4.1.1).

Phosphotransferase (EC 2.7): often called kinase, mostly ATP as the donor. For example, hexokinase, protein tyrosine kinase, etc. It should be noted that a few proteases are also called kinases (such as enterokinase), but they are hydrolases.

Hydrolase catalyzes the hydrolysis of substrates, such as proteases and lipases. Hydrolases generally play a degrading role and are mostly located outside the cell (such as in the digestive tract) or in the lysosome. Some proteases were also called kinases (such as enterokinase, EC 3.4.21.9, which is now called enteropeptidase).

Lyases catalyze the removal of a small molecule from the substrate, leaving a double bond (or ring) or its reverse reaction, such as aldolase, hydratase, decarboxylase, etc. IUBMB adjusted the classification and transferred part of the lyase to the transfer enzyme classification. For example, citrate synthase was once included in the lyase (EC 4.1.3.7) and has now been transferred to the acetyltransferase (EC 2.3.3.1).

Isomerases catalyze the mutual conversion between isomers, including racemase, epizyme, cis-trans isomerase, mutase (intramolecular group transfer) and intramolecular oxidation-reduction, Intramolecular elimination-addition reaction and other 6 subclasses.

Ligases catalyze the synthesis of one substance from two substances and must be coupled with ATP decomposition, such as DNA ligase, aminoacyl-tRNA synthetase, etc. Synthetase was used as a generic name before, because many people confused it with synthase, so IUBMB changed it to ligase in 1983.