Bio-auxiliaries

Method for producing sulphide tolerant bacterial proteases and its uses thereof
Disclosed herein is a method for producing sulphide-tolerant bacterial proteases and its uses in dehairing, hydrolysis of feathers and hair in leather processing applications. The method provides an approach for preparing an enzyme formulation from sulphide-tolerant Bacillus sp. (KLP1) and its application in enzyme-assisted dehairing of skins in leather processing applications. The patent application has been filed jointly with IIT Madras.

Indian Patent Application No. 201941006678

A metal resistant keratinase and a process for the preparation of keratin hydrolysate therewith
Disclosed herein is a keratinase of activity in the range of 710-5800 U/g, prepared from a bacterial source. An important feature of the keratinase is that it exhibits stability in the presence of metal ions, such as Arsenic, Nickel and Silver. Hence, 60-85% of the activity is retained even in the presence of the above metals. The keratinase finds application in the preparation of keratin hydrolysate, which is useful for the preparation of various value added products in different industrial sectors, e.g. cosmetic as well as leather industry. Indian Patent application no. 201811046312

A bacterial protease for dehairing of hide/skin and a process for the preparation thereof
Disclosed herein is a novel bacterial protease prepared from a Bacillus strain. The proteolytic activity of the enzyme is in the range of about 48,000-50,000 U/ml. It is alkaline in nature and is active at wide range of pH and temperature. This enzyme finds significant application in dehairing for the removal of hair in significantly reduced time without resulting in any sludge formation. Complete replacement of lime/sulphide with enzyme in the dehairing process leads to significant reduction of BOD, COD and TDS in the effluent without affecting the quality of the hides/skins. Thus, the protease ensures eco-benign beam house operation in leather making. Indian Patent application no. 201711019809.

An enzymatic composition and a process for the preparation thereof
Disclosed herein is an enzymatic composition produced from Aspergillus source by solid state fermentation. The enzymatic composition of the present invention finds potential application in leather processing industry to replace the hazardous chemicals conventionally used during pre-tanning operations. It also finds application in food industries for fruit clarifications. Further, it has potential application in pharmaceutical industries for green processing of drugs. Thus the enzymatic composition has potential application and ensuring effective replacement of synthetic chemicals, eco-benign option of processing and production of value added products from wastes. Indian Patent application no. is 201611004505.

A Novel Bioadhesive And A Process For The Preparation Thereof
Disclosed herein is a bioadhesive useful for both wet and dry tissue surfaces prepared using fibrous proteins. The protein is conjugated to simple bi-functional phenolics and then oxidized either with chemical or enzymatic oxidants. The resulting product acts as a resorbable adhesive and demonstrates antimicrobial property against both Gram +ve and Gram-ve bacteria and obviates the problems associated with surgical procedures. It finds potential application in clinical, surgical and pharmaceutical industries to ensure effective replacement of synthetic adhesives. It is useful in surgical practices (replacing the sutures, clips and staplers), intestinal defects, blood vessel damage, wound closure, tissue approximation and hemostasis.
Indian patent application no. is 1413Del2013.


A Process For The Production Of Bacterial Protease Enzyme For Industrial Application
Disclosed herein is a process for the preparation of bacterial protease from Bacillus subtilis MTCC5333. The bacterial protease finds application in leather processing industry, dairy industry, detergent industry, food processing industry, and also in the recovery of silver from photographic films.
Indian Patent application no. 1181Del2013.

A Novel Esterase and a Process for the Preparation thereof
A novel esterase exhibiting a feature of broad substrate specificity towards short-chain and long-chain p-nitrophenyl esters and triglycerides as well as stability in the presence of oxidants as well as surfactants is provided. Further the esterase has the potentiality to degrade three or more aliphatic polymers and co-polymers belonging to various groups. The esterase is prepared from a strain of Cryptococcus sp., designated as MTCC 5455, using solid state fermentation technique, wherein the production medium is prepared using locally available oil cakes. The process of preparation of the esterase does not need specific nitrogen or carbon source. The esterase finds potential application in detergent industry. It has also immense application potential in managing plastic wastes. The invention has been applied for patent protection (Indian Patent application no. 1937DEL2011).


A Novel Alkaline Protease for Industrial Applications and a Process for the Preparation thereof
A novel alkaline serine protease has been prepared from a strain of Bacillus pumilus under submerged culture condition. The said protease exhibits activity and stability in salts, surfactants, oxidants, detergent and various organic solvents. Further this novel protease possesses elastase activity but exhibits no collagenolytic activity. The protease finds potential application in leather processing. It also finds enormous industrial application for the purpose of cleaning and washing. The invention has been applied for patent protection (Indian Patent application no. 1939DEL2011).


A Novel Compound for Pharmaceutical Applications and a Process for the Preparation thereof
A novel compound for pharmaceutical applications has been derived from a marine microbial isolate by culturing the organism with defined substrates. The compound is extracted and purified under specified condition. The novel compound demonstrates anti-MRSA (Methicillin Resistant Staphylococcus aureus) activity and potential antioxidant activity when examined with standard free radical DPPH (Diphenyl-picridyl-hydrazine). It finds enormous application in pharmaceutical industry. The invention has been applied for patent protection (Indian Patent application no. 1578DEL2011).


A novel surface functionalised mesoporous activated carbon for industrial applications and a process for the preparation thereof
The invention provides a novel surface functionalized mesoporous activated carbon incorporated with selected functional groups, thereby enabling its attachment to a range of biomolecules. The product is envisaged to have enormous applications in the areas of biotechnology especially for the separation of enzymes from aqueous enzyme solution mixture. This also finds application in treatment of wastewater. The carbon matrix is also envisaged to have application in energy sector for the electrodes in fuel cell, bio fuel cells and in medical applications as a drug carrier surfaces. The invention further provides a simple process for the preparation of the surface functionalized mesoporous activated carbon.

A novel water-soluble bio-surfactant and a process for the preparation thereof
The present invention relates to an extra cellular Biosurfactant product obtained by growing the culture of Bacillus sp. and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5514, in production medium of containing peptic digested animal tissue, yeast extract and sodium chloride at pH 7.6 and incubated at 37C for 48h. Thus obtained biosurfactant product having exhibits: water-soluble in nature, both surface & emulsifying activity. And also exhibits stable emulsion up to 60 days.
The invention has been applied for patent protection (Indian Patent application no. 673Del2011).

A novel Collagenase and a process for the preparation thereof
The present invention relates to an extra cellular thermo stable enzyme obtained by growing the culture of Streptomyces sp. and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5211, in aqueous production medium containing essentially carbon source, nitrogen source and mineral source under controlled conditions. The collagenase exhibits activity at much wider range of pH and temperature, thereby assuring its application for many divergent industrial purposes. It is envisaged to have enormous potential application in chemical processing and pharmaceutical industries to ensure effective environmental management and product development. The enzyme is useful in leather industry for effective treatment of solid wastes generated by tanneries. It also finds medical applications especially for the purpose of development of biomaterials for wound management.
The invention has been applied for patent protection (Indian Patent application no. 125DEL2010).

A novel pH and thermostable tannase from a fungal source and a process for the preparation thereof
A novel pH and thermostable tannase is prepared from a novel isolated fungal stain of Aspergillus heteromorphous, isolated from soil samples of tea garden, Dibrugarh, Assam, India deposited in MTCC, IMTECH, Chandigarh and designated as MTCC 5466, by submerged culture fermentation, with activity higher than all other comparable reports in published literatures. This novel tannase exhibits activity and stability at wide range of pH (3.0-11.0) and temperature (15-90OC) besides showing stability towards organic solvents, oxidants, surfactants, denaturants with halotolerance property. The tannase finds potential application in treatment of vegetable tanning effluents from leather processing and as an additive in detergents industry to remove the tea stain.
The invention has been applied for patent protection (Indian Patent application no. 2372DEL2009).

A novel Bacteriocin and a process for the production thereof
A novel bacteriocin exhibiting a feature of antibacterial activity, antifungal activity and activity against the yeast strain and activity as well as stability in the wide range of pH and thermotolearnce as well as stability in organic solvents, chemicals and detergents is provided. While the bacteriocin exhibits antagonist activity, there is no collagenolytic activity. The broad spectrum bacteriocin I sprepared from a starin of Baciilus Lichenformis, designated as MTCC 5271, using submerged fermentation technique. Wherein the production medium is prepared with carbon and nitrogen source and the pH of the said medium is in the neutral range. The bacteriocin finds potential application in leather processing and food industries.
The invention has been applied for patent protection (Indian Patent application no. 781Del2010).

A novel alkaline protease and a process for the preparation thereof
A novel alkaline protease exhibiting a feature of halotolerance and activity as well as stability in the presence of oxidants as well as surfactants is provided. While the protease exhibits elastolytic activity, there is no collagenolytic activity. The alkaline protease is prepared from a strain of Bacillus subtilis, designated asMTCC 5333, using submerged fermentation technique, wherein the production medium is prepared without using any common salt or alkali and the pH of the said medium is the neutral range. The protease finds potential application in soaking as well as dehairing operations in leather processing industry. It may also be used as an additive in detergent industry.

The invention has been applied for patent protection (Indian Patent application no. 2375Del2007; Patent pending).


A process for the production of an alkaline protease
The process relates to the production of an extracellular alkaline thermo tolerant protease from a bacterial strain of genus Alcaligens (CLRI BL11). The process enables a much higher yield (800-1200 Anson units per ml of culture filtrate), compared to those in the hithertoknown processes. This protease exhibits stability at a pH in the range of 7-13 and a temperature of 50-65oC. The enzyme finds enormous application as a soaking and unhairing agent in leather processing industry. It also finds potential application in dairy industry for making cheese. Moreover, it may be used to prepare protein hydrolysates by controlling hydrolysis of proteins present in a substrate.

The invention has been applied for patent protection.(Indian Patent application no. 1122Del1999, Patent pending).


A process for the preparation of a proteolytic enzyme from a novel strain of fungus Paecilomyces lilacinus
The process relates to the production of a proteolytic enzyme from a fungal strain Paecilomyces lilacinus (MTCC 2835) using solid state fermentation technique. The enzyme exhibits activity at a pH in the range of 6-13 and temperature in the range of 25-70oC. The shelf life of the enzyme, unlike those of the conventional proteases, is as long as 2 years, when maintained at 5-20oC. The enzyme finds enormous application as bate, soaking agent as well as depilant in leather processing industry. It may also be used to treat leather industry byproducts such as fleshings, shavings etc to make value added products. Further, the enzyme may be used in meat industry, dairy, distillery, jute industry, food and fruit processing industry. It may also be used as an additive for enhancing efficacy of detergents.

The invention has been applied for patent protection.(Indian Patent application no. 377Del2000, Patent pending).


A process for the production of a novel lipase
The process relates to the production of lipase from a bacterial strain Ralstonia pickettii (CLRI BTL1). Unlike the conventional lipase, this enzyme exhibits stability over a wider range of pH (2-10) and temperature (4-80oC). The shelf life of the enzyme is around 90 days. The enzyme finds enormous application as a degreasing agent in leather processing industry. It may also be used as an additive to laundry detergent formulations. It has been found to be able to remove olive oil from cotton fabrics with better efficiency in the presence of available detergents. Moreover, it is a potential hydrolyzing agent for industrially important fats and oils, while producing fatty acids. It may find use in esterification, transesterification and interesterification reactions. It has also potential use as a digestive aid in pharmaceutical industry.

The invention has been applied for patent protection.(Indian Patent application no. 1555Del1999, Patent pending).


An improved process for the production of stable lipase
The process relates to the production of lipase from a fungal strain of Aspergillus niger (CLRI BTFL). Unlike the conventional lipase, this enzyme exhibits stability over a wider range of pH (2-11) and temperature (4-60oC). The enzyme finds enormous application as a degreasing agent in leather processing industry. It may also be used as an additive to laundry detergent formulations. It may also be used for the production of industrially important fatty acids from fats and oils in oleochemical industry. Chemical industries can use this enzyme for synthesis of esters and acylation of glucose. Moreover, it finds application in food and pharmaceutical industry for interesterification of butterfat, resolution of racemic mixtures and synthesis of peptides. It may even be used in the generation of dairy flavours for dairy industry.

The invention has been applied for patent protection.(Indian Patent application no. 1556Del1999, Patent pending).


A novel protease for industrial applications
The invention relates to an extracellular enzyme protease obtained by growing the culture of Pseudomonas aeruginosa isolated from vermiculture pit soil and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5270, in production medium of pH 7.0 and containing soybean meal and tryptone as raw materials, at 30°C for 72 h. Different agricultural products/ byproducts are used as protein sources. The main advantage of the invention is the use of simple substrates like soya flour, bengal gram flour and wheat bran for the production of protease.

The partially purified non-collagenolytic, calcium independent protease with molecular weight 60 kDa has activity in pH range of 6.0 – 11.0 and temperature range of 25-650C; stability in pH range of 6.0-10.0 and temperature 25-45°C. The protease activity is retained for 8 months when stored at ambient temperature.

The protease of the present invention finds enormous potential application in leather processing industry as an eco-benign option for dehairing of hides and skins with enzyme alone without supplementation of any chemical like lime or sodium sulphide. The use of protease reduces the dehairing time and makes dehairing more efficient resulting in better grain quality of leather. It may also be used in soaking process of leather manufacturing. The application of the enzyme avoids the pollution produced during pretanning operation associated with conventional method, thus making the enzyme, an ecofriendly product.

Further, the enzyme finds application as an additive in detergent and also for recovery of silver from waste X-ray and photographic films. It may also be used for preparations of protein hydrolysates. Yet another potential application of the protease of the present invention is in pharmaceutical industries and food processing industry.

The invention has been applied for Patent protection (Indian Patent application no. 2471DEL06).


A novel alkaline protease and a process for the production thereof
A novel alkaline protease has been produced from a Aspergillus source (MTCC5152). It is essentially an extra cellular alkaline protease active in a wide range of ph and temperature conditions. The invention further provides a process for the preparation of the novel protease using solid state fermentation or liquid fermentation. The extra cellular alkaline protease produced by the process of the present invention has potential.

Application in leather industry as a soaking and unhairing agent in the beamhouse processing. It may also be used for the purpose of food processing. The protease may also be used to prepare protein hydrolysates by controlling the hydrolysis of proteins present in a substrate. Being an alkaline protease, this enzyme may even be used as a supplement in laundry detergents.

The invention has been applied for Patent protection (Indian Patent application no. 0391DEL2007).

A novel enzymatic formulation for rapid fiber opening in leather making and process for the preparation thereof
Disclosed herein is a novel enzymatic formulation comprising galactosidase and serine protease prepared from a microbial source of Aspergillus terreus, designated as MTCC 5908. The formulation finds application in leather processing industry for the purpose of rapid opening of fiber bundles in animal hides and skins. The enzyme formulation can be used in leather processing at pH ranging from 4.0-12.0 without the addition of lime or any other solid carrier thereby reducing the BOD and COD of the effluent and the pollution thereof. Indian Patent application no. is 2357DEL2014.






For Further Information Please Contact

Head, Knowledge Portfolio Management Department (KPMD)

CSIR-Central Leather Research Institute,
Adyar, Chennai - 600 020
Tel: +91 44 24437273
E-mail: subhendu@clri.res.in