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Effect of the reversal of coenzyme specificity by expression of mutated Pichia stipitis xylitol dehydrogenase in recombinant Saccharomyces cerevisiae.
Lett Appl Microbiol. 2007 Aug; 45(2):184-9.LA

Abstract

AIMS

To determine the effects on xylitol accumulation and ethanol yield of expression of mutated Pichia stipitis xylitol dehydrogenase (XDH) with reversal of coenzyme specificity in recombinant Saccharomyces cerevisiae.

METHODS AND RESULTS

The genes XYL2 (D207A/I208R/F209S) and XYL2 (S96C/S99C/Y102C/D207A/I208R/F209S) were introduced into S. cerevisiae, which already contained the P. stipitis XYL1 gene (encoding xylose reductase, XR) and the endogenously overexpressed XKS1 gene (encoding xylulokinase, XK). The specific activities of mutated XDH in both strains showed a distinct increase in NADP(+)-dependent activity in both strains with mutated XDH, reaching 0.782 and 0.698 U mg(-1). In xylose fermentation, the strain with XDH (D207A/I208R/F209S) had a large decrease in xylitol and glycerol yield, while the xylose consumption and ethanol yield were decreased. In the strain with XDH (S96C/S99C/Y102C/D207A/I208R/F209S), the xylose consumption and ethanol yield were also decreased, and the xylitol yield was increased, because of low XDH activity.

CONCLUSIONS

Changing XDH coenzyme specificity was a sufficient method for reducing the production of xylitol, but high activity of XDH was also required for improved ethanol formation.

SIGNIFICANCE AND IMPACT OF THE STUDY

The difference in coenzyme specificity was a vital parameter controlling ethanolic xylose fermentation but the XDH/XR ratio was also important.

Authors+Show Affiliations

The State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

17651216

Citation

Hou, J, et al. "Effect of the Reversal of Coenzyme Specificity By Expression of Mutated Pichia Stipitis Xylitol Dehydrogenase in Recombinant Saccharomyces Cerevisiae." Letters in Applied Microbiology, vol. 45, no. 2, 2007, pp. 184-9.
Hou J, Shen Y, Li XP, et al. Effect of the reversal of coenzyme specificity by expression of mutated Pichia stipitis xylitol dehydrogenase in recombinant Saccharomyces cerevisiae. Lett Appl Microbiol. 2007;45(2):184-9.
Hou, J., Shen, Y., Li, X. P., & Bao, X. M. (2007). Effect of the reversal of coenzyme specificity by expression of mutated Pichia stipitis xylitol dehydrogenase in recombinant Saccharomyces cerevisiae. Letters in Applied Microbiology, 45(2), 184-9.
Hou J, et al. Effect of the Reversal of Coenzyme Specificity By Expression of Mutated Pichia Stipitis Xylitol Dehydrogenase in Recombinant Saccharomyces Cerevisiae. Lett Appl Microbiol. 2007;45(2):184-9. PubMed PMID: 17651216.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effect of the reversal of coenzyme specificity by expression of mutated Pichia stipitis xylitol dehydrogenase in recombinant Saccharomyces cerevisiae. AU - Hou,J, AU - Shen,Y, AU - Li,X P, AU - Bao,X M, PY - 2007/7/27/pubmed PY - 2007/10/17/medline PY - 2007/7/27/entrez SP - 184 EP - 9 JF - Letters in applied microbiology JO - Lett Appl Microbiol VL - 45 IS - 2 N2 - AIMS: To determine the effects on xylitol accumulation and ethanol yield of expression of mutated Pichia stipitis xylitol dehydrogenase (XDH) with reversal of coenzyme specificity in recombinant Saccharomyces cerevisiae. METHODS AND RESULTS: The genes XYL2 (D207A/I208R/F209S) and XYL2 (S96C/S99C/Y102C/D207A/I208R/F209S) were introduced into S. cerevisiae, which already contained the P. stipitis XYL1 gene (encoding xylose reductase, XR) and the endogenously overexpressed XKS1 gene (encoding xylulokinase, XK). The specific activities of mutated XDH in both strains showed a distinct increase in NADP(+)-dependent activity in both strains with mutated XDH, reaching 0.782 and 0.698 U mg(-1). In xylose fermentation, the strain with XDH (D207A/I208R/F209S) had a large decrease in xylitol and glycerol yield, while the xylose consumption and ethanol yield were decreased. In the strain with XDH (S96C/S99C/Y102C/D207A/I208R/F209S), the xylose consumption and ethanol yield were also decreased, and the xylitol yield was increased, because of low XDH activity. CONCLUSIONS: Changing XDH coenzyme specificity was a sufficient method for reducing the production of xylitol, but high activity of XDH was also required for improved ethanol formation. SIGNIFICANCE AND IMPACT OF THE STUDY: The difference in coenzyme specificity was a vital parameter controlling ethanolic xylose fermentation but the XDH/XR ratio was also important. SN - 0266-8254 UR - https://www.unboundmedicine.com/medline/citation/17651216/Effect_of_the_reversal_of_coenzyme_specificity_by_expression_of_mutated_Pichia_stipitis_xylitol_dehydrogenase_in_recombinant_Saccharomyces_cerevisiae_ L2 - https://doi.org/10.1111/j.1472-765X.2007.02165.x DB - PRIME DP - Unbound Medicine ER -