Title of Invention

A PROCESS FOR THE PREPARATION OF A NOVEL PROTEINOID FOR INDUSTRIAL APPLICATIONS

Abstract

The present invention provides a process for the preparation of novel proteinoid tanning material in the molecular weight range of 7000-10000 KD , which provides softness to leather.The process of present invention provides the preparation of novel proteinoid by means of oxidation of lysine group present in the poly peptide chain of proteinous material to aldehyde group to react chemically with reactive groups of said proteinous material. Said oxidation effected by treating the proteinous material with organo oxidizing agent selected from pyridinium chlorochromate, pyridinium bromochromate, piperidinium chlorochromate ,

Full Text

The present invention relates to a process for the preparation of a novel proteinoid for industrial applications. The novel proteinoid of the present invention has potential application in leather processing industry as a tanning agent to get softness, fullness, buffing property and lightweightness to the resulting leather. Moreover it may also be used in photographic and carpentry industry. Proteinoid is a material prepared from protein material either by acid or alkali hydrolysis. One of the important operations, the pickled pelts are subjected to during tanning operations with the help of various tanning auxiliaries to provide permanent stability to collagen matrix, against micro organism to convert putrescible skin into imputrescible leather. The tanning operation has a major role to play in connection with hydrothermal stability, tightness, fullness, lightweightness and softness of the final leather thereby increasing the hydrothermal stability, strength property of the resulting leather. Different tanning agents are conventionally used depending on the requirement of the properties and end uses of the final leather. Tanning agents have usually been of non-proteinous nature. But retanning agents of protein base have been already in use. Retanning agents are synthetically prepared which will behave like tanning agent to impart some of the properties such as grain tightness, softness and filling properties to the resulting leather. Ramamurthy et al (Journal of society of leather technologists and chemists.m 73,168,1985) provided a process for the preparation of keratin hydrolyzate to use as syntan in post tanning operation of leather processing. The keratin hydrolyzate is prepared by hydrolyzing keratin with acid or alkali by conventional process. The advantages of the keratin hydrolyzate are to increase the exhaustion of chrome to 85%-90% and also as a filling material in post tanning process. The major limitation associated in this hydrolyzate is that it cannot be used as a tanning agent because it donot react with reactive groups present in the collagen by forming any crosslinks there by not providing stability to the fibres and also decaying tendency of protein nature. The storage period keratin hydrolyzate is 6 months only. Another limitations associated with this syntan are that there is no improvement in grain tightness and softness. Our copending Indian Patent application No.210/DEL/2001 has provided a process for the preparation of proteinoid based acrylic syntan which has the wide application in leather processing. The syntan is prepared by hydrolyzing the protein source (flesh, chrome shaving, buffing dust and trimming) with acid or alkali and adjusting the pH of 6-7 and then subjected to coplymerisation with the help of acrylates. The advantages are that it can be used as a filling agent, grain tightening agent and can with stand the temperature of 150°C while at glazing. The syntan increases the exhaustion of dye to the level of 80-90% in the resulting leather. But the limitation associated with the syntan is that storage period of maximum 1 year and also not reacting with reactive groups by forming stable cross-links. The penetration of the same upto fibre level was also difficult because of the molecular weight range of 15000-20000 KD. Moreover it cannot be used as tanning agent because of decaying tendency of proteinous nature. Both keratin hydrolyzate and protein based acrylic syntan cannot be used as a tanning agent because the inability to preserve the leather and lack of softness, buffing property and fullness. The important limitation associated with keratin and protein based acrylic syntan was that it donot react with reactive groups present in collagen to form any cross link thereby providing permanent stability to the collagen fibres. The above limitations have prompted the researchers to look for better option in the realm of tanning agent from the proteinous source. The main object of the present invention is to provide a process for the preparation of novel proteinoid for industrial application, which obviates the drawback, stated above. Another object of the present invention is to provide a process for the preparation of novel proteinoid for tanning process to provide softness to the resulting leather. Yet another object of the present invention is to provide a process for the preparation of novel proteinoid for tanning process to provide fullness and buffing property to the resulting leather. Still another object of the present invention is to provide a process for the preparation of novel proteinoid tanning material in the molecular weight range of 7000-10000KD. Yet another object of the present invention is to provide a process for the preparation of novel proteinoid by means of oxidation of lysine group by using organo oxidizing agent present in the poly peptide chain of collagen to aldehyde group to react chemically with reactive groups of collagen. Accordingly the present invention provides a process for the preparation of a novel proteinoid tanning material having molecular weight in the range of 7000-10000 KD for industrial applications characterized in that treating the proteinous material with organo oxidizing agent selected from pyridinium chlorochromate, pyridinium bromochromate, piperidinium chlorochromate , said process comprises the step of, i) hydrolyzing proteinous material selected from fleshings, fibrin, keratin, elastin, collagen, for a period of 30 minutes followed by adjusting of pH of the hydrolyzate in the range of 6 - 7 and subsequent sterilization of the resulting hydrolyzate for a period in the range of 10 - 30 minutes, ii) separating the sterilized protein hydrolyzate, as formed in step (i) and adjusting the pH of the resulting solution in the range of 6 - 7 , iii) treating the hydrolyzate, as formed in step (ii), with 0.1% - 0.5% w/w, of organo oxidizing agent as defined above , in aqueous medium at temperature in the range of 4 - 12°C for a period of 24 - 72 hours to form oxidized proteinous hydrolyzate solution, iv) separating the proteinoid from the solution as formed in step (iii), by dialysis or resin adsorption under agitation for a period of 48 - 72 hours , at the temperature range of 4 - 12°C to obtain the novel proteinoid. 48-72 hours in the temperature range of 4-12°C to obtain the novel proteinoid. In an embodiment of the present invention, the protein material used may be selected from fleshings, fibrin, keratin, elastin, collagen. In another embodiment of the present invention, known method of sterilization used may be selected from the group consisting of ethylene oxide, gamma radiation, ultraviolet radiation In yet another embodiment of the present invention, known method of separation used may be selected from dialysis, resin absorption. In still another embodiment of the present invention, organo oxidising agent used may be selected from the group consisting of pyridinium chlorochromate, pyridinium bromochromate, piperidinium chlorochromate. In yet another enmbodiment of the present invention, the amount of oxidising agent used may be in the range of 0.1-0.5%w/v. The process of the present invention is described below in details. The proteinous source is taken and washed well to remove unwanted extraneous material. It is then subjected to conventional alkali or acid hydrolysis. Then the pH is adjusted to 6-7 by known method. The resulting mixture is sterilised by conventional method . Then the sterilised material is treated with organo oxidising agent in the range of 0.1-0.5%w/v for 24-72 hours at temperature range of 4-12°C to get oxidised proteinous material. The material Is separated by conventional method such as dialysis or resin adsorption to get novel proteinoid. Resin adsorption is basically an ion exchange phenomenon. Ion exchange resins, when placed in a solution, reach an equilibrium state between ions in solution and ions on the resin. The ions can then be selectively removed with help of suitable solvent system. For example cationic materials with the help of dilute alkalis. Dialysis is basically a desalting process in which excess salts formed during the previous operation in a protein mixture is removed by taking the material in a dialysis bag. The dialysis bag that we have used is commercially available and is made up of cellulose, which retains most proteins of molecular weight in the range of 12000 and greater. The novelty and non-obviousness of the present invention lies in the treatment of basic groups of lysine present in the proteinous source with organo oxidising agent at pH in the range of 6-7 thereby facilitating formation of cross link of the reactive group of the protein, resulting in not only in the improvement in stability but also providing softness, buffing property and light weightness property when it is used as tanning agent in leather processing. The following examples are given by way of illustration and therefore should not be construed to limit the scope of the present invention. Example 1 Dried and limed fleshing weighing 1kg was taken in a flask, washed with water to remove lime and other unwanted extraneous material. 100g sodium hydroxide was added to the washed material and hydrolyzed. The pH of the filtrate material around was 11. 3gm of hydrchloric acid was added to the hydrolysed material and the pH was adjusted to 6. Then the material was sterilised by UV radiation for 10 minutes. The hydrolyzate was then subjected to oxidation in the presence of 500 mg of pyridinium chloro chromate at an temperature of 4 °C for a period of 24 hours. The basic groups present in the hydrolyzed polypeptide proteins are converted into aldehydes to get proteinoid material. Then the proteinoid was seperated by resin absorption 50 ml of the proteinoid is taken and mixed with commercially available DOWEX ion exchange resin (pre-swelled with elution buffer). As the proteinoid material is cationic in nature, we have used dilute Hydrochloric acid as the elution buffer. The mixed material is packed on to a column of 50 cm length x 2cm width column tube and eluted with dilute Hydrochloric acid. The collected material is freeze dried in a lyophilizer. The resulting proteinoid material was used in leather processing as a tanning agent. Example 2 Fibrin weighing 1kg was taken in a flask, washed with water to remove unwanted extraneous material. 150g sodium hydroxide was added to the washed material and hydrolyzed. The pH of the filtrate material around was 11.5. 5gm of hydrchloric acid was added to the hydrolysed material the pH was adjusted to 7. Then the material was sterilised by gamma radiation for 20 minutes. The hydrolyzate was then subjected to oxidation in the presence of 300 mg of pyridinium bromo chromate at an temperature of 8°C for a period of 48 hours. The basic groups present in the hydrolysed polypeptide proteins 50 ml of the proteinoid is taken and mixed with commercially available DOWEX ion exchange resin (pre-swelled with elution buffer). As the proteinoid material is cationic in nature, we have used dilute Hydrochloric acid as the elution buffer. The mixed material is packed on to a column of 50 cm length x 2cm width column tube and eluted with dilute Hydrochloric acid. The collected material is freeze dried in a lyophilizer. The resulting proteinoid material was used in leather processing as a tanning agent. The resulting proteinoid material was used in leather processing as a tanning agent. Example 4 Collagen of pickled pelt weighing 1 kg was taken and washed thoroughly with brine solution to remove unwanted extraneous material. Then the pelts were depickled with 1 litre of water, 50 gm of salt and 10gm of sodium formate and the pH was adjusted to 4.5. Subsequently the depickled pelts were treated with 30gm of the proteinoid material and drummed for 3 hours for the penetration of tanning. Then 10 gm of sodium formate was added followed by 5gm of bicarbonate was given in three installments at 15 minutes interval for the fixation of tanning. Then the tanned leathers were aged for overnight and it was tested for the shrinkage temperature, fullness and softness. The leather showed softness, lightweightness and fullness. Advantages 1 The process of the present invention is very simple and involves no difficult parameter. 2.The process is very economical. 3. A process as claimed in calims 1-2 wherein hydrologises of proteinan material in carried out by treting with alkli hydroxide. 5.The novel proteinoid prepared from the protein source can be stored for more than 1 year. 4.The novel proteinoid prepared is able to react with reactive groups of collagen and forms stable cross links. 4.The novel proteinoid gives lightweightness to the resulting leather. 5.The leather produced by this tanning agent ensures good napp property to the suede leather. 6. The tanning agent prepared can be used as self-tanning material, results in softness and fullness. We Claim: 1. A process for the preparation of a novel proteinoid tanning material having molecular weight 7000-10000 KD for industrial applications characterized in that treating the proteinous material with organo oxidizing agent selected from pyridinium chlorochromate, pyridinium bromochromate, piperidinium chlorochromate , said process comprises the step of, i) hydrolyzing proteinous material selected from fleshings, fibrin, keratin, elastin, collagen, for a period of 30 minutes followed by adjusting of pH of the hydrolyzate in the range of 6 - 7 and subsequent sterilization of the resulting hydrolyzate for a period in the range of 10 - 30 minutes, ii) separating the sterilized protein hydrolyzate, as formed in step (i) and adjusting the pH of the resulting solution in the range of 6 - 7 , iii) treating the hydrolyzate, as formed in step (ii), with 0.1% - 0.5% w/w, of organo oxidizing agent as defined above , in aqueous medium at temperature in the range of 4 - 12°C for a period of 24 - 72 hours to form oxidized proteinous hydrolyzate solution, iv) separating the proteinoid from the solution as formed in step (iii), by dialysis or resin adsorption under agitation for a period of 48 - 72 hours , at the temperature range of 4 - 12°C to obtain the novel proteinoid. 2. A process as claimed in claims 1 - 2, wherein hydrolysis of proteinoid material is carried out by treating with alkali hydroxide . 3. A process as claimed in claims 1-3 wherein the pH of hydrolysate in step (i) & (ii) is adjusted by hydrochloric acid. 4. A process as claimed in claims 1 and 3, wherein method of sterilization used is selected from ethylene oxide, gamma radiation, ultraviolet radiation. 5. A process for the preparation of a novel proteinoid for industrial applications substantially as herein described with reference to the examples.

Documents:

172-del-2002-abstract.pdf

172-del-2002-claims.pdf

172-del-2002-correspondence-others.pdf

172-del-2002-correspondence-po.pdf

172-del-2002-description (complete).pdf

172-del-2002-form-1.pdf

172-del-2002-form-18.pdf

172-del-2002-form-2.pdf

172-del-2002-form-3.pdf