Expression and secretion of recombinant inulinase under the control of GAL or GAP promoter in Saccharomyces cerevisiae = Saccharomyces cerevisiae에서 GAL 또는 GAP 프로모터 조절에 의한 재조합 inulinase의 발현 및 분비
Cited 0 time in
- Expression and secretion of recombinant inulinase under the control of GAL or GAP promoter in Saccharomyces cerevisiae = Saccharomyces cerevisiae에서 GAL 또는 GAP 프로모터 조절에 의한 재조합 inulinase의 발현 및 분비
- Su Wan Nam; Hyun Houng Lim; Bong Hyun Chung; Yong Keun Chang
- Bibliographic Citation
- Korean Journal of Biotechnology and Bioengineering, vol. 11, no. 4, pp. 445-452
- Publication Year
- To investigate the promoter effect on heterologous gene expression in S. cerevisiae，the recombinant plasmids pYIll, pYI12, pYI10-2, and pYIGP were constructed to contain the inulinase gene (INU1、as a reporter under the control of GAL10, GAL7, GALI, and GAP promoters, respectively. When the yeasts transformants were cultivated on galactose-containing rich media, the cell growth reached to 36 -39 OD6oo at 72 hours of cultivation. The specific growth rates of the cells harboring the four different plasmids decreased similarily : they dropped from 0.24 h_l during the glucose-consuming period to 0.04 -0.10 h_, during the galactose-consuming period (gene expression phase for GAL promoter system). After the depletion of glucose, the expression of inulinase gene was started and reached to maximal levels of 4.3(GAL1 promoter), 4.0(GAL10 promoter), 3.S(GAL7 promoter), and 1.6(CL4P promoter) unit/mL at 72 hours of cultivation. Based on the maximal expression level and activity staining on the plate, the promoter strength was in the order of GALI, GAL10, GAL7 and GAP promoter. While the GAL-promoter systems showed a high plasmid stabilities of more than 78%, the GAP-promoter plasmid revealed a lower plasmid stability of 55%. Most of inulinase activity (98%) was found in the ex tracellular medium, indicating that the secretion efficiency of inulinase is independent on the type of promoter.
- Korea Soc-Assoc-Inst
- Appears in Collections:
- 1. Journal Articles > Journal Articles
- Files in This Item:
Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.