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Volume 153, Issue 9

Ethanol production from xylose by recombinant expressing protein-engineered NADH-preferring xylose reductase from Free

Abstract

A recombinant strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from (XR and XDH, respectively) has the ability to convert xylose to ethanol together with the unfavourable excretion of xylitol, which may be due to intercellular redox imbalance caused by the different coenzyme specificity between NADPH-preferring XR and NAD-dependent XDH. In this study, we focused on the effect(s) of mutated NADH-preferring XR in fermentation. The R276H and K270R/N272D mutants were improved 52- and 146-fold, respectively, in the ratio of NADH/NADPH in catalytic efficiency [( / with NADH)/( / with NADPH)] compared with the wild-type (WT), which was due to decrease of with NADPH in the R276H mutant and increase of with NADPH in the K270R/N272D mutant. Furthermore, R276H mutation led to significant thermostabilization in XR. The most positive effect on xylose fermentation to ethanol was found by using the Y-R276H strain, expressing XR R276H mutant and XDH WT: 20 % increase of ethanol production and 52 % decrease of xylitol excretion, compared with the Y-WT strain expressing XR WT and XDH WT. Measurement of intracellular coenzyme concentrations suggested that maintenance of the of NADPH/NADP and NADH/NAD ratios is important for efficient ethanol fermentation from xylose by recombinant .

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Keyword(s): AKR, aldo-keto reductase , CD, circular dichroism , CtXR, XR from Candida tenuis , PGK, phosphoglycerate kinase , PsXDH, XDH from Pichia stipitis , PsXR, XR from Pichia stipitis , Tm, thermal unfolding transition temperature , WT, wild-type , XDH, xylitol dehydrogenase , XI, xylose isomerase and XR, xylose reductase
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2007-09-01
2024-04-19
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Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing protein-engineered NADH-preferring xylose reductase from Pichia stipitis
Microbiology 153, 3044 (2007); https://doi.org/10.1099/mic.0.2007/007856-0
/content/journal/micro/10.1099/mic.0.2007/007856-0
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