Investigation of the Role of a Conserved Glycine Motif in the Saccharomyces cerevisiae Xylose Reductase

被引:0
|
作者
Byron C. H. Chu
Hung Lee
机构
[1] University of Guelph,Department of Environmental Biology
来源
Current Microbiology | 2006年 / 53卷
关键词
Xylose; Xylitol; Aldose Reductase; Xylose Reductase; Column Buffer;
D O I
暂无
中图分类号
学科分类号
摘要
All yeast xylose reductases, with the exception of that from Schizosaccharomyces pombe, possess the catalytic and coenzyme-binding elements from both the aldo–keto reductase and short-chain dehydrogenase–reductase (SDR) enzyme families in their primary sequences. In the Saccharomyces cerevisiae xylose reductase (XR), the SDR-like coenzyme-binding GXXXGXG motif (Gly motif) is located between residues 128 and 134, with the third Gly residue being replaced by an Asp. We used site-directed mutagenesis to study the role of this SDR-like Gly motif in the S. cerevisiae xylose reductase. Site-directed mutagenesis of the individual conserved Gly residue positions (G128A, G132A, D134G, and D134A) did not significantly affect the specific activity, kinetic constants (Km, Kcat, and Kcat/Km), or dissociation constants (Kd) in any of the variants compared with the wild type. Deletion of the entire Gly motif produced an unstable protein that could not be purified. These results indicate that the SDR-like Gly motif likely provides support to the overall structure of the enzyme, but it does not contribute directly to coenzyme binding in this XR.
引用
收藏
页码:118 / 123
页数:5
相关论文
共 50 条
  • [1] Investigation of the role of a conserved glycine motif in the Saccharomyces cerevisiae xylose reductase
    Chu, Byron C. H.
    Lee, Hung
    CURRENT MICROBIOLOGY, 2006, 53 (02) : 118 - 123
  • [2] Mutational study of the role of tyrosine-49 in the Saccharomyces cerevisiae xylose reductase
    Jeong, EY
    Sopher, C
    Kim, IS
    Lee, H
    YEAST, 2001, 18 (11) : 1081 - 1089
  • [3] Effect of glucose on xylose utilization in Saccharomyces cerevisiae harboring the xylose reductase gene
    Ji-Hye Han
    Ju-Yong Park
    Kye Sang Yoo
    Hyun Woo Kang
    Gi-Wook Choi
    Bong-Woo Chung
    Jiho Min
    Archives of Microbiology, 2011, 193 : 335 - 340
  • [4] Effect of glucose on xylose utilization in Saccharomyces cerevisiae harboring the xylose reductase gene
    Han, Ji-Hye
    Park, Ju-Yong
    Yoo, Kye Sang
    Kang, Hyun Woo
    Choi, Gi-Wook
    Chung, Bong-Woo
    Min, Jiho
    ARCHIVES OF MICROBIOLOGY, 2011, 193 (05) : 335 - 340
  • [5] On the Role of a Balanced Cofactor Recycling between Xylose Reductase and Xylitol Dehydrogenase in Recombinant Xylose Fermenting Strains of Saccharomyces cerevisiae
    Krahulec, S.
    Longus, K.
    Klimacek, M.
    Nidetzky, B.
    JOURNAL OF BIOTECHNOLOGY, 2010, 150 : S12 - S12
  • [6] The effect of xylose reductase genes on xylitol production by industrial Saccharomyces cerevisiae in fermentation of glucose and xylose
    Yang, Bai-Xue
    Xie, Cai-Yun
    Xia, Zi-Yuan
    Wu, Ya-Jing
    Li, Bo
    Tang, Yue-Qin
    PROCESS BIOCHEMISTRY, 2020, 95 : 122 - 130
  • [7] High activity of xylose reductase and xylitol dehydrogenase improves xylose fermentation by recombinant Saccharomyces cerevisiae
    Karhumaa, Kaisa
    Fromanger, Romain
    Hahn-Hagerdal, Barbel
    Gorwa-Grauslund, Marie-F.
    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2007, 73 (05) : 1039 - 1046
  • [8] High activity of xylose reductase and xylitol dehydrogenase improves xylose fermentation by recombinant Saccharomyces cerevisiae
    Kaisa Karhumaa
    Romain Fromanger
    Bärbel Hahn-Hägerdal
    Marie-F. Gorwa-Grauslund
    Applied Microbiology and Biotechnology, 2007, 73 : 1039 - 1046
  • [9] Comparison of the xylose reductase-xylitol dehydrogenase and the xylose isomerase pathways for xylose fermentation by recombinant Saccharomyces cerevisiae
    Kaisa Karhumaa
    Rosa Garcia Sanchez
    Bärbel Hahn-Hägerdal
    Marie-F Gorwa-Grauslund
    Microbial Cell Factories, 6
  • [10] Comparison of the xylose reductase-xylitol dehydrogenase and the xylose isomerase pathways for xylose fermentation by recombinant Saccharomyces cerevisiae
    Karhumaa, Kaisa
    Garcia Sanchez, Rosa
    Hahn-Hagerdal, Barbel
    Gorwa-Grauslund, Marie-F
    MICROBIAL CELL FACTORIES, 2007, 6 (1)