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Title 

Enhanced secretion of Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae by translational fusion to cellulose-binding domain

Authors 

Jungoh AhnEui Sung ChoiHong-Weon LeeS H HwangC S KimH W JangS J HaamJ K Jung

Publisher 

Springer Verlag (Germany)

Issue Date 

2004

Citation 

Applied Microbiology and Biotechnology, vol. 64, no. 6, pp. 833-839

Keywords 

cellulosetriacylglycerol lipasebinding affinitygeobacillus stearothermophilusprotein domainprotein secretionsaccharomyces cerevisiaebacillus stearothermophilusbinding siteslipase

Abstract 

The secretion of Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae was investigated by employing a fusion partner, a cellulose-binding domain (CBD) from Trichoderma harzianum endoglucanase II (THEG). The CBD was connected to the N-terminal of L1 lipase through an endogenous linker peptide from THEG. The expression cassette for the fusion protein in S. cerevisiae was constructed using the α-amylase signal peptide and the galactose-inducible GAL10 promoter. Secretion of CBD-linker-L1 lipase by this fusion construct was dramatically 7-fold enhanced, compared with that of the mature L1 lipase without CBD-fusion. The fusion protein was secreted into the culture medium, reaching levels of approximately 1.3 g/l in high-cell-density fed- batch cultures. Insertion of a KEX2 cleavage site into the junction between CBD-linker and L1 lipase resulted in the same level of enhanced secretion, indicating that the CBD- linker fusion probably plays a critical role in secretion from endoplasmic reticulum to Golgi apparatus. Therefore, the CBD from THEG can be used both as an affinity tag and as a secretion enhancer for the secretory production of heterologous proteins in S. cerevisiae, since in vivo breakage at the linker was almost negligible.

ISSN 

0175-7598

Link 

http://dx.doi.org/10.1007/s00253-003-1547-5

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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