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Title 

Cloning of a gene encoding dextranase from Lipomyces starkeyi and its expression in Pichia pastoris

Authors 

H K KangJi-Young ParkJ S AhnS H KimD Kim

Publisher 

The Korean Society for Applied Microbiology

Issue Date 

2009

Citation 

Journal of Microbiology and Biotechnology, vol. 19, no. 2, pp. 172-177

Keywords 

dextrandextranaseexpressionlipomyces starkeyipichia pastoris

Abstract 

A gene (lsd1) encoding dextranase from Lipomyces starkeyi KSM22 has been previously cloned, sequenced, and expressed in Saccharomyces cerevisiae. The gene consisting of 1,824 base pairs and encoding a protein of 608 amino acids was then cloned into and secretively expressed in Pichia pastoris under the control of the AOX1 promoter. The dextranase productivity of the P. pastoris transformant (pPIC9K-LSD1, 134,000 U/l) was approximately 4.2-fold higher than that of the S. cerevisiae transformant (pYLSD1, 32,000 U/l) cultured in an 8-1 fermentor. Over 0.63 g/l of active dextranase was secreted into the medium after methanol induction. The dextranase of the P. pastoris transformant, as analyzed by SDS-PAGE and Western blotting, showed only one homogeneous band. This dextranase of the P. pastoris transformant showed a broad band near 73 kDa. Rabbit monoclonal antibodies against a synthetic LSD1 peptide mix also recognized approximately 73 kDa.

ISSN 

1017-7825

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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