Title of Invention	"A PROCESS FOR THE PRODUCTION OF BIODEGRADABLE FILM FROM MICROBIAL SOURCES"
Abstract	In the invention Pseudomonas caryophilli CFR 1705 bacteria has been cultured in whey, a byproduct of dairy industry, as carbon source to induce formation of polysaccharide for the production of biodegradable film to be used as packaging material. The polysaccharide produced by the said microorganism are very specific in their physico-chemical characteristics and functional properties.

Full Text

The present invention relates to a process for the production of biodegradable film from microbial sources. The process in particular uses polysaccharide from Pseudomonas caryophilli CFR 1705 as microbial source. The novel aspect of the present invention is that, the polysaccharide produced is used to make biodegradable films. Petrochemical based plastics such as polyethylene, polyamides, polystyrene and polyolefens are being used to displace traditional packaging materials such as tin cans, glass containers, paperboard, etc,. Though the plastics have made significant inroads in packaging they are currently under attack from environmental point of view. A new realization has dawned in both producers and consumers about the disposal of plastics. As plastics are petroleum based long chain hydrocarbons, they cannot be degraded by biological system such as fungi and bacteria. Disposal of plastic is a global problem. Incineration is one of the method of waste which results in toxic fumes in the environment which can pose a serious pollution problem. Recalcitrant plastics accumulate in the environment at a rate of 25 million tons each year. The fate of these polymers in the environment and the time required for their total mineralization to carbon dioxide have yet to be fully understood. Biodegradable polymers offer an attractive alternative to the petroleum based non biodegradable polymers. Search for biodegradable polymers is on. Reference may be made to the work of Peter D. Hoagland and Nicholas Parris (1996), who have carried out work on chitosan - animal origin films. They made chitosan and pectin laminated films by using glycerol or lactic acid as plastisizers. Reference may also be made to the work of Garcia M.A., Martino M.N., and Zartzky (2000). They have formulated starch based films from plant sources. They have used glycerol and sorbitol as the plasticizers. Reference may be made to the work of Coffin,D.R., Fishman, M.L., (US patent 5,646,206) carried out work on the fabrication of films from mixture of pectin and polyvinyl alcohol. Reference may be made to the work of Fishma, M.L., Coffin, D.R., (US patent 5,451,673) who have carried out work on the fabrication of films from mixture of pectin and starch. hi all the above cited references the source of obtaining the film uses from animal or plant origin where as the present invention is based on microbial source. The main objective of this invention is to provide a process for the production of biodegradable film from microbial sources. Novelty of the process is that , the polysaccharide produced by Pseudomonas caryophilli CFR 1705 has the ability to form into biodegradable films . Unlike other biobased films such as starch, pectin and chitin, it is not cost prohibitive and uses whey, a byproduct as the carbon source. The polysaccharides produced by microorganisms are very specific in their physico- chemical characteristics and functional properties. This particular strain CFR1705 is unique in its performance in producing polysaccharide based on lactose/whey as a carbon source besides its film forming capability. Accordingly the present invention provides a process for the production of biodegradable films from microbial sources which comprises, i) growing the Pseudomonas caryophilli CFR 1705 bacteria in a conventional media such as herein described followed by culturing in whey for a period of 72 hrs at 28 ° C at 200 rpm rotation to get culture broth, ii) pasteurizing the culture broth of step (i) at 60°C for 15 minutes, iii) precipitating polysaccharide from pasteurized broth using solvent as herein described , iv) washing of precipitate with acetone v) drying the precipitate in conventional drier for 8 to 10 hrs vi) grinding of dried polysaccharide material for a mesh size of 40 to 60 vii) dissolving the ground material of step (vi) in water at ambient temperature for a period of 45 min to 1 hour with vigorous stirring viii) heating the dissolved polysaccharide solution to 100°C for a minimum period of 20 minutes ix) cooling the solution to ambient temperature and spread the solution over a rigid sheet for a minimum thickness of 100 micron x) drying of film to a moisture content of 8 to 10% using mechanicak dryer to obtain desired biodegradable films. In an embodiment of the present invention, the organism used may be from Pseudomonas caryophilli CFR 1705. This culture is deposited at CFTRI, culture center and cab be made available to the public as per the adopted international norms to provide access of these cultures to the interested member of public. In an another embodiment of the present invention, biodegradable material may be produced having the film forming characteristics. Yet another embodiment of the present invention, biodegradable film may be prepared using the polysaccharide produced from Pseudomonas caryophilli CFR1705. Soil samples collected from different habitats are screened for the organism producing polysaccharide, on agar plate containing sucrose-2%, Na2HPO4-0.5%, KH2PO4-0.3%,MgSO4-0.1%, NaCL-0.1%,CaCl2 - 0.01% tryptone- 0.25%, yeast extract-0.1%.coloniy with slimy appearance is isolated and transferred on sterilised agar medium of the same composition and repeated several times. The purified isolate cultivated in a sterilised medium composed of sucrose-3%, Na2HPO4 - 0.5%, KH2PO4-0.3%, MgS04-0.1%, Nad- 0.1%, CaCl2- 0.01%, tryptone the morphological and biochemical studies according to the eighth edition of Bergy's manual and was identified as Pseudomonas caryophilli later numbered as CFR 1705. In the preliminary trials lactose was found to be preferred carbon source over the other carbohydrate sources such as sucrose, glucose, maltose, starch, cellulose, and pectin. The polysaccharide elaborated is found to be higher with an efficiency of 60 to 70%. The the industrial waste from the dairy industry especially the whey was found to favour the polysaccharide production by P.caryophilli CFR 1705. The yield of polysaccharide was comparable to that of pure lactose indicating better utilisation of whey as a carbon source. The following examples are given with illustration of present invention: EXAMPLE 1 This example illustrates the isolation and identification of polysaccharide producing organism. In this example the medium used with the composition of sucrose-2%, Na2HP04 - 0.5%, KH2P04 - 0.3%, MgSO4 -0.1%, NaCl - 0.!%, CaCl2 - 0.01%, Tryptone - 0.25%, yeast extract - 0.1% was sterilized by means of autoclaving at 151b for 20 minutes. The sterilized media are poured on to sterilized petriplates. The soils from different sources are screened by serial dilution and pour plate method. Petriplates were incubated at 37° C for 24 hrs. slimy colony was isolated. Isolated colony was identified based on microbiological and biochemical tests. (Table-l& Fig-1). Table-1 Cultural characteristics of Pseudomonas caryophilli CFR 1705 (Table Removed) EXAMPLE 2 This example illustrates the production of polysaccharide from Pseudomonas caryophilli CFR1705. In this example the medium used with the composition of lactose-3%,peptone-0.25%, sodium phosphate dibasic-0.5%, potassium phosphate-0.3%, magnesium sulphate-0.1%, sodium chloride-0.1%, yeast extract-0.1%, calcium chloride-0.01%, was sterilized by means of autoclaving at 151b for 20 minutes. The sterilized media were inoculated with 10% inoculum of 24 hrs. inoculated flasks were incubated for the period of 72hrs at ambient temperature on a rotary shaker with 200rpm. EXAMPLE-3 This example illustrates the production of polysaccharide form whey, hi this example whey was used as medium with adjustment of pH to 7. Whey (100ml) was taken in 500ml Erlenmeyer flasks and sterilized by means of autoclaving at 151b for 20 minutes. The sterilized media were inoculated with 10% inoculum of 24hrs old culture. The flasks were incubated for the period of 72hrs at 28°C on a rotary shaker with 200 'rpm EXAMPLE 4 After 72 hrs the culture broth is pasteurized at 60°C for 15 minutes and polysaccharide was precipitated with 1:2 volume of isopropyl alcohol. The precipitate was washed with acetone and dried in an oven for 8 to 10 hrs. Then the polysaccharide is ground to a mesh size of 40 to 60. The powder is stored in a bottle at ambient temperature. (Fig-2) EXAMPLE 5 This example illustrate the production of film from polysaccharide. In this example different concentration of polysaccharide is dissolved in water at ambient temperature. The polysaccharide solution is heated to a minimum period of 20 minutes on water bath. Heated solution is cooled and spread over a rigid sheet. Sheet is leveled with leveler to get uniform thickness of the film. The films were dried for 8to 10 hrs in an hot air oven. The dried films were peeled off from the plate and stored in an ambient temperature.(Fig-3) The main advantages of the present invention are 1. A process for the production of polysaccharide, which can be made into biodegradable films, from microbial sources. 2. Utilisation of cheaper carbon source. We Claim: 1. A process for the production of biodegradable films from microbial sources which comprises, i) growing the Pseudomonas caryophilli CFR I705 bacteria in a conventional media such as herein described followed by culturing in whey for a period of 72 hrs at 28 ° C at 200 rpm rotation to get culture broth, ii) pasteurizing the culture broth of step (i) at 60°C for 15 minutes, iii) precipitating polysaccharide from pasteurized broth using solvent as herein described , iv) washing of precipitate with acetone v) drying the precipitate in conventional drier for 8 to 10 hrs vi) grinding of dried polysaccharide material for a mesh size of 40 to 60 vii)dissolving the ground material of step( vi) in water at ambient temperature for a period of 45 min to 1 hour with vigorous stirring viii) heating the dissolved polysaccharide solution to 100°C for a minimum period of 20 minutes ix) cooling the solution to ambient temperature and spread the solution over a rigid sheet for a minimum thickness of 100 micron x) drying of film to a moisture content of 8 to 10% using mechanical laier to obtain desired biodegradable films. 2. A process as claimed in claim 1 wherein the biodegradable material is produced by precipitating polysaccharides from culture broth using isopropyl alcohol as solvent. 3. A process for the production of biodegradable film from microbial sources substantially as herein described with reference to the examples and figures accompanying this specification.

Documents:

440-del-2001-abstract.pdf

440-del-2001-claims.pdf

440-del-2001-correspondence-others.pdf

440-del-2001-correspondence-po.pdf

440-del-2001-description (complete).pdf

440-del-2001-drawings.pdf

440-del-2001-form-1.pdf

440-del-2001-form-18.pdf

440-del-2001-form-2.pdf

440-del-2001-form-3.pdf