Abstract:
:Xylose reductase catalyzes the NAD(P)H-dependent reduction of xylose to xylitol and is essential for growth on xylose by yeasts. To understand the nature of coenzyme binding to the Pichia stipitis xylose reductase, we investigated the role of the strictly conserved Lys270 in the putative IPKS coenzyme binding motif by site-directed mutagenesis. The Lys270Met variant exhibited lower enzyme activity than the wild-type enzyme. The apparent affinity of the variant for NADPH was decreased 5-16-fold, depending on the substrate used, while the apparent affinity for NADH, measured using glyceraldehyde as the substrate, remained unchanged. This resulted in 4.3-fold higher affinity for NADH over NADPH using glyceraldehyde as the substrate. The variant also showed a 14-fold decrease in Km for xylose, but only small changes were observed in Km values for glyceraldehyde. The wild-type enzyme, but not the Lys270Met variant, was susceptible to modification by the Lys-specific pyridoxal 5'-phosphate. Results of our chemical modification and site-directed mutagenesis study indicated that Lys270 is involved in both NADPH and D-xylose binding in the P. stipitis xylose reductase.
journal_name
FEMS Microbiol Lettjournal_title
FEMS microbiology lettersauthors
Kostrzynska M,Sopher CR,Lee Hdoi
10.1111/j.1574-6968.1998.tb12848.xsubject
Has Abstractpub_date
1998-02-01 00:00:00pages
107-12issue
1eissn
0378-1097issn
1574-6968pii
S0378-1097(97)00555-7journal_volume
159pub_type
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1998-03-15 00:00:00
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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更新日期:1999-10-15 00:00:00
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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更新日期:2005-08-15 00:00:00
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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更新日期:2007-10-01 00:00:00
abstract::It is now well established that physicochemical properties of exopolysaccharides (EPS) can vary between strains of a given species and according to growth conditions. The EPS production of four strains of Lactobacillus bulgaricus was monitored during growth in milk and in a chemically defined media. All strains, inclu...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
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更新日期:2003-04-25 00:00:00
abstract::Atypical enteropathogenic Escherichia coli (aEPEC) is comprised of a large heterogeneous group of strains and serotypes that carry the intimin gene (eae) but no other EPEC virulence factors. In a previous study, we examined a few aEPEC strains of O157:H16 serotype from the U.S. and France and found these to be nearly ...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
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更新日期:2012-12-01 00:00:00
abstract::Using pulsed-field gel electrophoresis (PFGE), nine profile types were identified in XbaI digests of a panel of strains of Salmonella enteritidis phage type 4 (PT 4) isolated in England and Wales between 1967 and 1992. When applied to 39 isolates from humans, chickens, poultry products and feed, strains belonging to p...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1994.tb06888.x
更新日期:1994-06-01 00:00:00