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Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae.
Appl Microbiol Biotechnol. 2008 Nov; 81(2):243-55.AM

Abstract

A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis has the ability to convert xylose to ethanol together with the unfavorable excretion of xylitol, which may be due to cofactor imbalance between NADPH-preferring XR and NAD(+)-dependent XDH. To reduce xylitol formation, we have already generated several XDH mutants with a reversal of coenzyme specificity toward NADP(+). In this study, we constructed a set of recombinant S. cerevisiae strains with xylose-fermenting ability, including protein-engineered NADP(+)-dependent XDH-expressing strains. The most positive effect on xylose-to-ethanol fermentation was found by using a strain named MA-N5, constructed by chromosomal integration of the gene for NADP(+)-dependent XDH along with XR and endogenous xylulokinase genes. The MA-N5 strain had an increase in ethanol production and decrease in xylitol excretion compared with the reference strain expressing wild-type XDH when fermenting not only xylose but also mixed sugars containing glucose and xylose. Furthermore, the MA-N5 strain produced ethanol with a high yield of 0.49 g of ethanol/g of total consumed sugars in the nonsulfuric acid hydrolysate of wood chips. The results demonstrate that glucose and xylose present in the lignocellulosic hydrolysate can be efficiently fermented by this redox-engineered strain.

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Authors+Show Affiliations

Matsushika A
Biomass Technology Research Center, National Institute of Advanced Industrial Science and Technology, 2-2-2 Hirosuehiro, Kure, Hiroshima, 737-0197, Japan. a-matsushika@aist.go.jp
Watanabe S
No affiliation info available
Kodaki T
No affiliation info available
Makino K
No affiliation info available
Inoue H
No affiliation info available
Murakami K
No affiliation info available
Takimura O
No affiliation info available
Sawayama S
No affiliation info available

MeSH

Aldehyde ReductaseD-Xylulose ReductaseEthanolNADPPichiaRecombinant ProteinsSaccharomyces cerevisiaeXylose

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18751695

Citation

Matsushika, Akinori, et al. "Expression of Protein Engineered NADP+-dependent Xylitol Dehydrogenase Increases Ethanol Production From Xylose in Recombinant Saccharomyces Cerevisiae." Applied Microbiology and Biotechnology, vol. 81, no. 2, 2008, pp. 243-55.
Matsushika A, Watanabe S, Kodaki T, et al. Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae. Appl Microbiol Biotechnol. 2008;81(2):243-55.
Matsushika, A., Watanabe, S., Kodaki, T., Makino, K., Inoue, H., Murakami, K., Takimura, O., & Sawayama, S. (2008). Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae. Applied Microbiology and Biotechnology, 81(2), 243-55. https://doi.org/10.1007/s00253-008-1649-1
Matsushika A, et al. Expression of Protein Engineered NADP+-dependent Xylitol Dehydrogenase Increases Ethanol Production From Xylose in Recombinant Saccharomyces Cerevisiae. Appl Microbiol Biotechnol. 2008;81(2):243-55. PubMed PMID: 18751695.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae. AU - Matsushika,Akinori, AU - Watanabe,Seiya, AU - Kodaki,Tsutomu, AU - Makino,Keisuke, AU - Inoue,Hiroyuki, AU - Murakami,Katsuji, AU - Takimura,Osamu, AU - Sawayama,Shigeki, Y1 - 2008/08/27/ PY - 2008/06/16/received PY - 2008/08/03/accepted PY - 2008/07/31/revised PY - 2008/8/30/pubmed PY - 2008/12/17/medline PY - 2008/8/30/entrez SP - 243 EP - 55 JF - Applied microbiology and biotechnology JO - Appl Microbiol Biotechnol VL - 81 IS - 2 N2 - A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis has the ability to convert xylose to ethanol together with the unfavorable excretion of xylitol, which may be due to cofactor imbalance between NADPH-preferring XR and NAD(+)-dependent XDH. To reduce xylitol formation, we have already generated several XDH mutants with a reversal of coenzyme specificity toward NADP(+). In this study, we constructed a set of recombinant S. cerevisiae strains with xylose-fermenting ability, including protein-engineered NADP(+)-dependent XDH-expressing strains. The most positive effect on xylose-to-ethanol fermentation was found by using a strain named MA-N5, constructed by chromosomal integration of the gene for NADP(+)-dependent XDH along with XR and endogenous xylulokinase genes. The MA-N5 strain had an increase in ethanol production and decrease in xylitol excretion compared with the reference strain expressing wild-type XDH when fermenting not only xylose but also mixed sugars containing glucose and xylose. Furthermore, the MA-N5 strain produced ethanol with a high yield of 0.49 g of ethanol/g of total consumed sugars in the nonsulfuric acid hydrolysate of wood chips. The results demonstrate that glucose and xylose present in the lignocellulosic hydrolysate can be efficiently fermented by this redox-engineered strain. SN - 1432-0614 UR - https://www.unboundmedicine.com/medline/citation/18751695/Expression_of_protein_engineered_NADP+_dependent_xylitol_dehydrogenase_increases_ethanol_production_from_xylose_in_recombinant_Saccharomyces_cerevisiae_ L2 - https://dx.doi.org/10.1007/s00253-008-1649-1 DB - PRIME DP - Unbound Medicine ER -