Cloning and Overexpression of Gene Encoding the Pullulanase from Bacillus naganoensis in Pichia pastoris
Cloning and Overexpression of Gene Encoding the Pullulanase from Bacillusnaganoensis in Pichia pastoris
- Resource Type
- Article
- Authors
- XU, BO; YUN-JUAN YANG; ZUN-XI HUANG
- Source
- Journal of Microbiology and Biotechnology, 16(8), pp.1184-1191 Aug, 2006
- Subject
- 생물학
- Language
- ISSN
- 1017-7825
The expression of a pullulanase gene in Pichiawas investigated. The gene encoding pullulanase wascloned by PCR using the chromosomal DNA of Bacilusnaganoensis as the template. The expression vector pPIC9K-Puwas constructed by inserting the pullulanase gene into plasmidpPIC9K and then transformed into Pichia pastoris SMD168by electroporation. Activity determination, SDS-PAGE, andPCR amplification indicated that the gene of the pululanase fromB. naganoensisand the molecular size of the expressed recombinant productwas about 119.9 kDa. This is the first report on the successfulexpression of the pullulanase from B. naganoensis in P.pastoris. The transformant secreted recombinant pululanasewith the activity of 350.8 IU/ml in shake-flask culture. Theproperties of the recombinant pululanase were characterized.