Cloning and expression in Saccharomyces cerevisiae of the NAD(P)H-dependent xylose reductase-encoding gene (XYL1) from the xylose-assimilating yeast Pichia stipitis

Amore, René; Kötter, Peter; Küster, C; Ciriacy, Michael; Hollenberg, Cornelis P.

doi:10.1016/0378-1119(91)90592-y

Cited by 138 publications

(84 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The amino acid sequence of the P . stipitis xylose reductase indicates the presence of three cysteine residues per subunit (Amore et al, 1991 ; Hallborn et al, 1991 ; Takuma et al, 1991) and the order of reaction determined from our study suggests that one of these residues was modified to effect inactivation.…”

Section: Inactivation Of Xylose Reductase By Pmbs and Dtnbmentioning

confidence: 50%

“…Although xylose reductases have been purified from several yeasts (Bolen et al, 1985(Bolen et al, , 1986Ditzelmuller et al, 1984;Ho et al, 1990;Rizzi et al, 1988;Scher & Horecker, 1966;Suzuki & Onishi, 1975;Verduyn et al 1985;Watson et al, 1969) and the amino acid sequence of the P. stipitis xylose reductase has recently been published (Amore et al, 1991;Hallborn et al, 1991;Takuma et al, 1991), little is known about the enzyme's catalytic mechanism or the nature of its catalytic sites. Yeast xylose reductases have been classified as members of the aldose reductase enzyme family (alditol : NAD(P)+ 1-oxidoreductase, EC 1 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of xylose reductase from the yeast Pichia stipitis: Evidence for functional thiol and histidyl groups

Webb

Lee

1992

Journal of General Microbiology

View full text Add to dashboard Cite

Xylose reductase (EC 1.1.1.21) from the yeast Pichia stipitis NRC 2548 was purified to homogeneity via a twostep protocol using anion-exchange and gel-filtration chromatography. The pH-activity profile revealed the presence of two ionizable groups with pK,,, values of 5.8 and 8.1, suggesting the catalytic involvement of histidyl and thiol groups, respectively. Additional evidence supporting the involvement of these residues was provided by the use of group-specific inhibitors. The enzyme was rapidly inactivated in a pseudo-first order manner by the thiolspecific modifier p-chloromercuriphenylsulphonate (PMBS) and analysis of the order-of-reaction suggested that one essential cysteine residue was modified to effect inactivation. Treatment of the enzyme with another thiolspecific modifier, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), showed that modification of one cysteine per monomer led to 90 % loss of activity, further supporting the existence of one essential cysteine residue. Inactivation by PMBS was reversed by adding 1 mM-P-mercaptoethanol. Inactivation of xylose reductase by the histidinespecific modifier diethylpyrocarbonate (DEP) followed a pseudo-first order process, and analysis of the order-ofreaction suggested that one essential histidine residue was modified to effect inactivation. Treatment of DEPinactivated enzyme with 0.2 M-neutral hydroxylamine resulted in the recovery of 45% of enzyme activity. Protection of xylose reductase from PMBS-and DEP-inactivation was provided by NADPH and NADH but not by NADP+, D-XylOSe or Dbglyceraldehyde. This suggests that the essential histidine and cysteine residues may be involved with binding of cofactor by the P(, stipitis xylose reductase.

show abstract

Section: Inactivation Of Xylose Reductase By Pmbs and Dtnbmentioning

confidence: 50%

Section: Introductionmentioning

confidence: 99%

Characterization of xylose reductase from the yeast Pichia stipitis: Evidence for functional thiol and histidyl groups

Webb

Lee

1992

Journal of General Microbiology

View full text Add to dashboard Cite

show abstract

“…For maximum ethanol production, it requires a system where the reduction of acetaldehyde under limited oxygen conditions is possible, and additionally oxidation of xylitol to xylulose [80]. The capability of S. cerevisiae to assimilate and utilize xylose was clearly observed with the isolation and transformation of two XR genes (XYL1 and XYL2) from P. stipites, which were responsible for the assimilation of xylose [141,142]. The genetically engineered S. cerevisiae with those two genes had the ability to utilize xylose by oxidation processes and generate xylitol in the absence of any additional co-metabolizable carbon sources for xylose assimilation processes.…”

Section: Pathway Of Xylose Reductase and Xylitol Dehydrogenasementioning

confidence: 99%

Bioethanol a Microbial Biofuel Metabolite; New Insights of Yeasts Metabolic Engineering

et al. 2018

View full text Add to dashboard Cite

Scarcity of the non-renewable energy sources, global warming, environmental pollution, and raising the cost of petroleum are the motive for the development of renewable, eco-friendly fuels production with low costs. Bioethanol production is one of the promising materials that can subrogate the petroleum oil, and it is considered recently as a clean liquid fuel or a neutral carbon. Diverse microorganisms such as yeasts and bacteria are able to produce bioethanol on a large scale, which can satisfy our daily needs with cheap and applicable methods. Saccharomyces cerevisiae and Pichia stipitis are two of the pioneer yeasts in ethanol production due to their abilities to produce a high amount of ethanol. The recent focus is directed towards lignocellulosic biomass that contains 30-50% cellulose and 20-40% hemicellulose, and can be transformed into glucose and fundamentally xylose after enzymatic hydrolysis. For this purpose, a number of various approaches have been used to engineer different pathways for improving the bioethanol production with simultaneous fermentation of pentose and hexoses sugars in the yeasts. These approaches include metabolic and flux analysis, modeling and expression analysis, followed by targeted deletions or the overexpression of key genes. In this review, we highlight and discuss the current status of yeasts genetic engineering for enhancing bioethanol production, and the conditions that influence bioethanol production.

show abstract

“…X59465) and XYL2 (GenBank accessioin no. AF127801) [9,10]. The PCR reaction was performed in 25-ll reaction mixtures (0.15 lM each primer, 1 ll of template DNA [about 10 ng of genomic DNA], 12.5 ll PCR premix [Invitrogen 403061]).…”

Section: Strains and Mediamentioning

confidence: 99%

Metabolic engineering of Saccharomyces cerevisiae for increased bioconversion of lignocellulose to ethanol

Cai

2012

Indian J Microbiol

View full text Add to dashboard Cite

The absence of pentose-utilizing enzymes in Saccharomyces cerevisiae is an obstacle for efficiently converting lignocellulosic materials to ethanol. In the present study, the genes coding xylose reductase (XYL1) and xylitol dehydrogenase (XYL2) from Pichia stipitis were successfully engineered into S. cerevisae. As compared to the control transformant, engineering of XYL1 and XYL2 into yeasts significantly increased the microbial biomass (8.1 vs. 3.4 g/L), xylose consumption rate (0.15 vs. 0.02 g/h) and ethanol yield (6.8 vs. 3.5 g/L) after 72 h fermentation using a xylose-based medium. Interestingly, engineering of XYL1 and XYL2 into yeasts also elevated the ethanol yield from sugarcane bagasse hydrolysate (SUBH). This study not only provides an effective approach to increase the xylose utilization by yeasts, but the results also suggest that production of ethanol by this recombinant yeasts using unconventional nutrient sources, such as components in SUBH deserves further attention in the future.

show abstract

Cloning and expression in Saccharomyces cerevisiae of the NAD(P)H-dependent xylose reductase-encoding gene (XYL1) from the xylose-assimilating yeast Pichia stipitis

Cited by 138 publications

References 40 publications

Characterization of xylose reductase from the yeast Pichia stipitis: Evidence for functional thiol and histidyl groups

Characterization of xylose reductase from the yeast Pichia stipitis: Evidence for functional thiol and histidyl groups

Bioethanol a Microbial Biofuel Metabolite; New Insights of Yeasts Metabolic Engineering

Metabolic engineering of Saccharomyces cerevisiae for increased bioconversion of lignocellulose to ethanol

Contact Info

Product

Resources

About