Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Atypon full text link Atypon Free PMC article
Full text links

Actions

Share

.1998 May;64(5):1852-9.
doi: 10.1128/AEM.64.5.1852-1859.1998.

Genetically engineered Saccharomyces yeast capable of effective cofermentation of glucose and xylose

Affiliations

Genetically engineered Saccharomyces yeast capable of effective cofermentation of glucose and xylose

N W Ho et al. Appl Environ Microbiol.1998 May.

Abstract

Xylose is one of the major fermentable sugars present in cellulosic biomass, second only to glucose. However, Saccharomyces spp., the best sugar-fermenting microorganisms, are not able to metabolize xylose. We developed recombinant plasmids that can transform Saccharomyces spp. into xylose-fermenting yeasts. These plasmids, designated pLNH31, -32, -33, and -34, are 2 microns-based high-copy-number yeast-E. coli shuttle plasmids. In addition to the geneticin resistance and ampicillin resistance genes that serve as dominant selectable markers, these plasmids also contain three xylose-metabolizing genes, a xylose reductase gene, a xylitol dehydrogenase gene (both from Pichia stipitis), and a xylulokinase gene (from Saccharomyces cerevisiae). These xylose-metabolizing genes were also fused to signals controlling gene expression from S. cerevisiae glycolytic genes. Transformation of Saccharomyces sp. strain 1400 with each of these plasmids resulted in the conversion of strain 1400 from a non-xylose-metabolizing yeast to a xylose-metabolizing yeast that can effectively ferment xylose to ethanol and also effectively utilizes xylose for aerobic growth. Furthermore, the resulting recombinant yeasts also have additional extraordinary properties. For example, the synthesis of the xylose-metabolizing enzymes directed by the cloned genes in these recombinant yeasts does not require the presence of xylose for induction, nor is the synthesis repressed by the presence of glucose in the medium. These properties make the recombinant yeasts able to efficiently ferment xylose to ethanol and also able to efficiently coferment glucose and xylose present in the same medium to ethanol simultaneously.

PubMed Disclaimer

Figures

FIG. 1
FIG. 1
Construction of high-copy-number yeast-E. coli shuttle plasmids pLNH31, pLNH32, pLNH33, and pLNH34. containing theXYL three-gene cassette KK-AR (or A*R)-KD. TheXhoI DNA fragment containing KK-AR (or A*R)-KD was inserted into pUCKm10 at itsSalI site.
FIG. 2
FIG. 2
Construction ofE. coli plasmids containing theXYL gene cassette KK-AR (or A*R)-KD. KK,S. cerevisiae XK gene fused to the promoter of theS. cerevisiae pyruvate kinase gene; AR (or A*R),P. stipitis XR gene fused to the promoter of theS. cerevisiae alcohol dehydrogenase gene; KD,P. stipitis XDH gene fused to the promoter of theS. cerevisiae pyruvate kinase gene. The double dagger (‡) indicates that theXhoI site was regenerated after ligation, and the asterisks (∗) indicate an intact ADC1 promoter.
FIG. 3
FIG. 3
Growth ofS. cerevisiae in rich medium with or without a carbon source (sugar). (A)S. cerevisiae AH22 cultured in YEP or YEPD. (B)S. cerevisiae AH22 cultured in YEP or YEPXylu (1% yeast extract, 2% peptone, 2% xylulose). Symbols: ○, YEP; •, YEPD; ▴, YEPXylu.
FIG. 4
FIG. 4
Comparison of the abilities of recombinantSaccharomyces sp. strain 1400(pLNH32) and parent strain 1400 to coferment glucose and xylose. (A) Strain 1400(pLNH32). (B) Strain 1400. Utilization of glucose and xylose and production of ethanol, xylitol, and glycerol were determined. Symbols: ▪, glucose; •, xylose; ▴, ethanol; □, xylitol; ▵, glycerol.
FIG. 5
FIG. 5
Fermentation of xylose by recombinantSaccharomyces sp. strain 1400(pLNH32) cultured in the xylose-containing medium YEPX. Utilization of xylose and production of ethanol, xylitol, and glycerol were determined. Symbols: •, xylose; ▴, ethanol; □, xylitol; ▵, glycerol.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ammerer G. Expression of genes in yeast using the ADC1 promoter. Methods Enzymol. 1983;101:192–201. - PubMed
    1. Armstrong K A, Som T, Volkert F C, Rose A, Broach J R. Propagation and expression of genes in yeast using 2-micron circle vectors. In: Barr P J, Brake A J, Valenzuela P, editors. Yeast genetic engineering. Boston, Mass: Butterworths; 1989. pp. 165–192. - PubMed
    1. Becker D, Guarente L. High efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–186. - PubMed
    1. Bennetzen J L, Hall B D. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase I. J Biol Chem. 1982;257:3018–3025. - PubMed
    1. Bolen P L, Detroy R W. Induction of NADPH-linked d-xylose reductase and NAD-linked xylitol dehydrogenase activities in Pachysolen tannophilus by d-xylose, l-arabinose, or d-galactose. Biotechnol Bioeng. 1985;27:302–307. - PubMed

Publication types

MeSH terms

Substances

Related information

LinkOut - more resources

Full text links
Atypon full text link Atypon Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp