A Novel Whole-Cell Biocatalysis Using In Production Of Biodiesel Of Jatropha Curcas

Biocatalysis is the major research field in the transesterification of biodiesel. Discoverying new lipase and developing the whole-cell biocatalysis may be the key problems in biocatalysis of biodiesel. Jatropha curcas is an important lipids resourse of plant, which is valuable in biodiesel. In this paper the lipase of J. curcas was cloned and was expressed in Pichia farinosa to make a novel whole cell biocatalysis. The effect of the biocatalysis was then studied. This will provide a practical performance for the exploitation of Jatropha.The lipase of J. curcas (JcLIP3) was cloned for the first time by gene cloning methods based on bioinformatics. The full-length cDNA was obtained by RACE experiment, which was 1582bp and encoded 356aa protein in its fullest ORF. The gene was expressed in E. coli to characterize the function. The enzyme activity was 0.8U/mL in E. coli. The bioinformatics analysis results showed that JcLIP protein hads the same activity centre as that of Rhizomucor miehei, but a different lid structure from the known structure.In this paper the expression system of Pichia farinosa was first constructed. The structure of integration vector, method of preparation of competent cells and parameters of electroporation was determined. And the genetic stability of this system was tested and results showed that it could be stable in 20 days' continuous culture. The JcLIP3 gene was then expressed in this system. The results of RT-PCR, protein expression analysis and determination of enzyme...