|
KMID : 0613820170270121403
|
|
Journal of Life Science
2017 Volume.27 No. 12 p.1403 ~ p.1409
|
|
|
Expression System for Optimal Production of Xylitol Dehydrogenase (XYL2) in Saccharomyces cerevisiae
|
|
Jung Heo-Myung
Kim Yeon-Hee
|
|
|
Abstract
|
|
|
|
In this study, the xylitol dehydrogenase (XYL2) gene was expressed in Saccharomyces cerevisiae as a host cell for ease of use in the degradation of lignocellulosic biomass (xylose). To select suitable expression systems for the S.XYL2 gene from S. cerevisiae and the P.XYL2 gene from Pichia stipitis, pGMFα-S.XYL2, pGMFα-P.XYL2, pAMFα-S.XYL2 and pAMFα-P.XYL2 plasmids with the GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor α (MFα) signal sequence was also connected to each promoter to allow secretion. Each plasmid was transformed into S. cerevisiae SEY2102△trp1 strain and the xylitol dehydrogenase activity was investigated. The GAL10 promoter proved more suitable than the ADH1 promoter for expression of the XYL2 gene, and the xylitol dehydrogenase activity from P. stipitis was twice that from S. cerevisiae. The xylitol dehydrogenase showed NAD+-dependent activity and about 77% of the recombinant xylitol dehydrogenase was secreted into the periplasmic space of the SEY2102△trp1/pGMFα-P.XYL2 strain. The xylitol dehydrogenase activity was increased by up to 41% when a glucose/xylose mixture was supplied as a carbon source, rather than glucose alone. The expression system and culture conditions optimized in this study resulted in large amounts of xylitol dehydrogenase using S. cerevisiae as the host strain, indicating the potential of this expression system for use in bioethanol production and industrial applications.
|
|
|
KEYWORD
|
|
|
Pichia stipitis, promoter strength, Saccharomyces cerevisiae, secretion production, xylitol dehydrogenase
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|