Stable Gene Variants of Lipases
The present technology encompasses methods for generating variation in gene sequences (of E. coli Lipase), provides protocols for screening the enzymes with higher thermostability and also protocols for sequencing and expression. The method according to present technology includes four steps: Ist step – To generate variation in the primary sequence of the lipase gene by PCR methods 2nd step - The mutant sequences are cloned into an expression vector and the protein expressed in the culture lysates. The expressed proteins are screened for their ability to withstand higher temperature was tested in a large population using medium-throughput methods. 3rd step - The promising variants are pooled by a family shuffling procedure and further tested for thermostability. In addition to the shuffling procedures selective changes in the primary sequence were also incorporated by standard molecular biology procedures. 4th step - The positive sequences are over-expressed in large cultures and proteins are purified. The purified proteins are tested for their thermostability.
Sector: Medical Technologies
Area of Application: Biotechnology, Food industry, Pharmaceutical industry
Keywords: lipases, gene variants, sequencing, expression, mutant
Advantages: This Method give access to production of thermostable lipases. These tailored thermostable lipases designed for high temperature use and thus improves the overall performance production process Tailored Lipases improve the yield and reduces the cost in bioreactors. Fit to use for production of a wide variety of foods, especially from milk, fat and oil. Lipases finds use in production of detergents, pharmaceuticals, perfumes, flavor enhancers and texturising agents in cosmetics products. Ubiquitous enzymes of considerable physiological significance and industrial potential
Environmental aspects: Not Applicable
Development Status: Laboratory Model
Legal Protection: Patent
Transfer Terms: Technology Licensing
Target Countries: India
Estimated cost (US$):
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