1a. Objectives (from AD-416)
The objective of the collaborative work is to isolate and express the lipase gene(s), and then proceed to characterize the enzymatic properties of the gene-products from Candida bombicola yeast used in the production of the biosurfactant sophorolipid.
1b. Approach (from AD-416)
Using a polymerase-chain-reaction gene-fishing approach developed and routinely used in our laboratory, the lipase gene(s) will be cloned from Candida bombicola – a yeast species that produces sophorolipid biosurfactant. The genes will be expressed in E.coli or other suitable host, and the enzymes will be purified and characterized by following common methodologies. Their application potential in the synthesis of nutraceuticals through interesterification reactions and in the formulation of washing mixes will be assessed.
3. Progress Report
Project researchers (PRs) carried out computer program-aided alignment of known pseudomonas lipase sequences and, based on the data, designed degenerative oligonucleotide-primers to use in a subsequent successful PCR-cloning of lipase and its modulator genes from polyhydroxyalkanoate-synthesizing bacterium, Pseudomonas resinovorans. In silico sequence analyses were performed to characterize the identity and structural properties of the gene products. Study is in progress to verify the function of the gene products, and to subclone the genes in E. coli for expression and large-scale production of the lipase for testing in food-related applications. Progress of the research project was monitored through frequent face-to-face meetings with the visiting scientist while on-site, and e-mail contacts and teleconference calls.