Title of Invention	"A PROCESS FOR RECOVERY OF BIOSURFACTANT FROM DISTILLERY WASTE"
Abstract	A process for recovery of biosurfactant from fermented distillery waste by adsorbing biosurfactant obtained from fermented distillery waste on wood activated carbon (WAC), recovering the biosurfactant by disorption of biosurfactant by organic solvent selected from ethanol, methanol, acetone, diethylether, and buffer removing solvent to recover colour free biosurfactant

Full Text

The present invention relates to a process for the recovery of biosurfactant produced from distillery waste by adsorption-desorption process process using activated carbon. More specifically, the invention relates to the recovery of colourfree biosurfactatn from fermented distillery waste through the following protocol given in scheme. Biosurfactants are beginning to acquire a status as potential performance-effective molecules in various field as an ecofriendly alternative to synthetic surfactants. Their range of potential industrial applications includes enhanced oil recovery, crude oil drilling, lubricants, surfactants-aided bioremediation of heavy metal and water-insoluble pollutants. In view of wide-spread utilization, process development for large scale production of biosurfactant is an obvious necessity. Therefore, it is of interest to the manufacturing industries to develop the biosurfactant production in which the cost of the raw material and the process are minimal. Hence, it is necessary to give an insight into the economic strategies which emphasizes on utilization of waste streams as no cost substrate and low cost in situ recovery methods for biosurfactant. Both of these objectives are essential for developing a large scale production technology of biosurfactant. In search of cheaper raw material for biosurfactant, use of industrial wastes have shown good promise. Studies on bench scale have shown that biosurfactant production was comparable in distillery and wey wastes and in synthetic medium (Babu. P.S.I vaidya A. N., Bal, A. S., Kapur, R., Juwarkar, A. . and Khanna, P., 1996, Kenetics of biosurfactant production by Pseudomonas aeruginosa strain BS2 from industrial waste. Biotechnology Letters, vol. 18, pp. 263-268; Dubey, K. and Juwarkar, A.I 2001, Distillery and curd whey as viable alternative sources for biosurfactant production. World Journal of Microbiology and Biotechnology, vol. 17, pp. 61-69). It is well known that most of the fermentation products are released in dilute aqueous solutions. Therefore, in many cases down stream processing in many biotechnological processes is responsible for upto 60% of total production cost. Due to economic considerations, most biosurfactant would have to involve either whole-cell spent culture broths or other crude preparations in some of the industries such as oil industry which requires lower purity specifications. However, the cosmetics, food and pharmaceutical industries are low volume, high-value categories which could absorb the higher cost of biosurfactants. Uptil now the methods used for biosurfactant recovery include solvent extraction, precipitation, crystallization, centrifugation and foam fractionation. However, these methods cannot be used when distillery waste is used as no cost complete nutrient medium for biosurfactant production because recovery of biosurfactant by any one of these classic method imparts colour to the biosurfactant making it non aesthetic in appearance. Moreover, these methods cannot be used for continuous product recovery during cultivation. Besides these methods, there are some methods for biosurfactant recovery which can be called in situ recovery in which the products are continuously removed from the culture broth during cultivation. In situ methods for product recovery be of increasing use and adopted in future because they avoid end product inhibition during fermentation, reduce costs of solvent and wastewater treatment, and minimize product degradation. Some examples for in situ biosurfactant recovery are adsorption of the surfactant onto ion exchange resin or other suitable adsorbents as in the case of lipopeptide surfactant produced by Candida petrophilum (Iguchi, T., takeda, I. and Ohsawa, H. 1969, Agricultural and Biological Chemistry, vol. 33, pp. 1657). Due to their lipophilic ability, the rhamnolipid could be adsorbed onto a support, which is termed XAD-2 followed by purification and freeze drying which resulted in only 60% recovery of biosurfactant with 90% purity (Reiling, H.F., Wyass, U.T., Guerra-Santos, I.H., Hirt, R., Kappeli, O. and Fiecheter, A., 1986, Pilot plant production of rhamnolipid biosurfactant by Pseudomonas aeruginosa. Applied and Environmental Microbiology, vol. 81, pp. 985-989). One of the main disadvantages of these exchangers when used for recovery of organic macromolecules in that the delicate structure of the molecules is disrupted and the product cannot be recovered without denaturation. Additionally adsorption capacity is generally small. One of the most successful techniques involved a continuous in situ removal of surfactin biosurfactant produced by Bacillus subtilis from fermentation broth by foam fractionation. In this technique foam was collected and acidified. To pH - 2 with concentrated HCI and the precipitated surfactant was extracted in dichloromethane (Copper, D.G., MacDonald , C.R., Dull, S.J.B. and Kosaric, N., 1981, Enhanced production of surfactin from B. subtilis by continuous product removal and metal cation additions. Applied and Environmental Microbiology, vol. 42, pp. 408-412). However, use of this method is not appropriate in case of biosurfactant removal from fermented distillery waste owing to interferences of colour impurities in the recovered product. Hence, main objective of the present invention is to provide a new process for recovery of biosurfactant from distillery waste by adsorption-desorption process using activated carbon, which obviates the above-described drawbacks. Also the other objective of the present invention is to explore the feasibility of biosurfactant revovery without having colour impurities from distillery waste which is used as a viable no-cost complete nutrient medium for biosurfactant production by P. aeruginosa given accession No. BS2 by screening of a suitable adsorbent among several such as silica gel, activated alumina, wood activated carbon, and zeolite so that selective adsorption and recovery of biosurfactant from fermented distillery waste is feasible. Still another object of the present invention is to demonstrate the usefulness of wood activated carbon (WAC) which is screened as an efficient adsorbent for biosurfactant recovery from fermented distillery waste through optimization of various parameters which affects adsorption of biosurfactant on the carbon in batch mode. Yet another object of the present invention is to demonstrate the feasibility of biosurfactant removal from collapsed foam (collected by foam fractionation method) which has 5 folds higher concentration of biosurfactant than that present in fermented distillery waste by adsorption-desorption process. Yet another objective of the present invention is to elute the adsorbed biosurfactant by using suitable eluant in a colourfree form and to assess the improvement in the surface active characteristic of the recovered biosurfactant based on critical micelle concentration determination. Still yet another objective of the present invention is the demonstration of usefulness of activated carbon through scanning electron microscopic studies to obtain colourless biosurfactant mediated by adsorption-desorption process. Accordingly the present invention provides a process for recovery of biosurfactant from fermented distillery waste which comprises: a) characterized in that adsorbing biosurfactant obtained from fermented distillery waste on wood activated carbon (WAC), b) recovering the biosurfactant by disorption of biosurfactant by organic solvent selected from ethanol, methanol, acetone, diethylether, and buffer. c) removing solvent to recover colour free biosurfactant. The present invention provides a new process for recovery of biosurfactant from distillery waste by adsorption-desorption process using activated carbon which comprises of: screening of a suitable adsorbent for adsorption of biosurfactant from fermented distillery waste demonstration of usefulness of activated carbon for adsorption of biosurfactant from fermented waste through optimization of various parameters which affects adsorption of biosurfactant from fermented distillery waste studying the feasibility of removing biosurfactant by adsorption on to activated carbon from collapsed foam in which biosurfactant is concentrated by 5 folds. Screening of suitable eluant for desorption of colourfree biosurfactant from activated carbon Demonstration of usefulness of WAC for adsorption-desorption of biosurfactant from fermented waste by SEM studies Pseudomonas aeruginosa strain BS2, a potential biosurfactant producing culture was isolated at NEERI from an oily sludge (Dubey, K. and Juwarkar, A., 2001, Distillery and curd whey as viable alternative sources for biosurfactant production. World Journal of Microbiology and Biotechnology, vol. 17, pp. 61- 69). Batch fermentation was carried out in New Bruswick BIO FLO II C fermenter using (1:3) diluted distillery spent wash as nutrient medium for proliferation of the strain and biosurfactant production under following conditions i.e. temperature 37°C, agitation 400 ppm, incubation period 96 hours and aeration at 2 SLPM. 1 and 1/2 WP (volume per air per volume liquid). After 96 hours the fermented waste was made cell free (by centrifugation at 8000 rpm and 0.45 um membrane filtration ) and 0.2% v/v formaldehyde was added to present microbial growth. The fermented cell free waste was analysed for pH surface tension (ST), Fcmc i.e. dilution factor to reach critical micelle concentration and biosurfactant yield Claim: 1. A process for recovery of biosurfactant from fermented distillery waste which comprises: a) characterized in that adsorbing biosurfactant obtained from fermented distillery waste on wood activated carbon (WAC), b) recovering the biosurfactant by disorption of biosurfactant by organic solvent selected from ethanol, methanol, acetone, diethylether, and buffer. c) removing solvent to recover colour free biosurfactant. 2. A process as claimed in claim 1 wherein the distillery waste is obtained from fermentation of distillery waste by Pseudomonal aeruginosa strain BS2 or bacillus subtilis by known batch fermentation method or foam fermentation method. 3. A process as claimed in claims 1-2, wherein the activated carbon screened as the best adsorbent among silicagel, zeolite-A and activated alumina. 4. A process as claimed in claims 1 - 3, wherein the contact time for biosurfactant adsorption is varying from 10-19 min. 5. A process as claimed in claims 1 - 4, wherein the carbon dose is ranging between from 0.2-0.4 % w/v. 6. A process as claimed in claims 1 - 5, wherein the volume of the fermented distillery waste is varying from 10-100 ml keeping the carbon dose constant (0.5g). 7. A process as claimed in claims 1 - 6, wherein the adsorption of biosurfactant is carried out at a temperature ranging between 30 - 50°C keeping carbon dose constant at 1% w/v. 8. A process as claimed in claims 1 -7 wherein, adsorption of biosurfactant is carried out at varied pH ranging between 3-10. 9. A process as claimed in claims 1 -8 wherein WAC dose is ranging from 1-6% for removal of biosurfactant from collapsed foam in which biosurfactant is concentrated by 5 folds. 10. A process as claimed in claims 1-9 wherein recovered biosurfactant had much lower critical micelle concentration 0.013 mg/ml indicating no loss in its structure and improvements surface active property. 11. A process for recovery of biosurfactant from fermented distillery waste substantially as herein described with reference to the examples.

Documents:

351-DEL-2002-Abstract-(06-11-2008).pdf

351-del-2002-abstract.pdf

351-DEL-2002-Claims-(06-11-2008).pdf

351-del-2002-claims.pdf

351-DEL-2002-Correspondence-Others-(06-11-2008).pdf

351-del-2002-correspondence-others.pdf

351-del-2002-correspondence-po.pdf

351-DEL-2002-Description (Complete)-(06-11-2008).pdf

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

351-del-2002-form-1.pdf

351-del-2002-form-18.pdf

351-DEL-2002-Form-2-(06-11-2008).pdf

351-del-2002-form-2.pdf

351-del-2002-form-3.pdf