Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis
A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis (PsXR and PsXDH, respectively) has the ability to convert xylose to ethanol together with the unfavourable excretion of xylitol, which may be due to interc...
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| Format: | Article |
| Language: | English |
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CSIRO Publishing
2007
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| Online Access: | http://ir.unimas.my/id/eprint/13992/ http://ir.unimas.my/id/eprint/13992/1/Seiya.pdf |
| _version_ | 1848837541048877056 |
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| author | Seiya, Watanabe Abu Saleh, Ahmed Narayana, Visu Annaluru |
| author_facet | Seiya, Watanabe Abu Saleh, Ahmed Narayana, Visu Annaluru |
| author_sort | Seiya, Watanabe |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis (PsXR and PsXDH, 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 PsXR in fermentation. The R276H and K270R/N272D mutants were improved 52- and 146-fold, respectively, in the ratio of NADH/NADPH in catalytic efficiency [(kcat/Km with NADH)/(kcat/Km with NADPH)] compared with the wild-type (WT), which was due
to decrease of kcat with NADPH in the R276H mutant and increase of Km with NADPH in the K270R/N272D mutant. Furthermore, R276H mutation led to significant thermostabilization in PsXR. The most positive effect on xylose fermentation to ethanol was found by using the
Y-R276H strain, expressing PsXR R276H mutant and PsXDH WT: 20% increase of ethanol production and 52% decrease of xylitol excretion, compared with the Y-WT strain expressing
PsXR WT and PsXDH 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 S. cerevisiae. |
| first_indexed | 2025-11-15T06:41:18Z |
| format | Article |
| id | unimas-13992 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:41:18Z |
| publishDate | 2007 |
| publisher | CSIRO Publishing |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-139922021-07-12T15:40:25Z http://ir.unimas.my/id/eprint/13992/ Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis Seiya, Watanabe Abu Saleh, Ahmed Narayana, Visu Annaluru TP Chemical technology A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis (PsXR and PsXDH, 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 PsXR in fermentation. The R276H and K270R/N272D mutants were improved 52- and 146-fold, respectively, in the ratio of NADH/NADPH in catalytic efficiency [(kcat/Km with NADH)/(kcat/Km with NADPH)] compared with the wild-type (WT), which was due to decrease of kcat with NADPH in the R276H mutant and increase of Km with NADPH in the K270R/N272D mutant. Furthermore, R276H mutation led to significant thermostabilization in PsXR. The most positive effect on xylose fermentation to ethanol was found by using the Y-R276H strain, expressing PsXR R276H mutant and PsXDH WT: 20% increase of ethanol production and 52% decrease of xylitol excretion, compared with the Y-WT strain expressing PsXR WT and PsXDH 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 S. cerevisiae. CSIRO Publishing 2007 Article PeerReviewed text en http://ir.unimas.my/id/eprint/13992/1/Seiya.pdf Seiya, Watanabe and Abu Saleh, Ahmed and Narayana, Visu Annaluru (2007) Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis. Microbiology Australia, 153. pp. 3044-3054. ISSN 1324-4272 https://www.researchgate.net/publication/6072782 DOI: 10.1099/mic.0.2007/007856-0 ยท |
| spellingShingle | TP Chemical technology Seiya, Watanabe Abu Saleh, Ahmed Narayana, Visu Annaluru Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title | Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title_full | Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title_fullStr | Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title_full_unstemmed | Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title_short | Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing proteinengineered NADH-preferring xylose reductase from Pichia stipitis |
| title_sort | ethanol production from xylose by recombinant saccharomyces cerevisiae expressing proteinengineered nadh-preferring xylose reductase from pichia stipitis |
| topic | TP Chemical technology |
| url | http://ir.unimas.my/id/eprint/13992/ http://ir.unimas.my/id/eprint/13992/ http://ir.unimas.my/id/eprint/13992/ http://ir.unimas.my/id/eprint/13992/1/Seiya.pdf |