Efficient, galactose-free production of Candida antarctica lipase B by GAL10 promoter in Δgal80 mutant of Saccharomyces cerevisiae

Whang, Jake; Ahn, Jungoh; Chang-Soo, Chun; Son, Young Soo; Lee, Hongweon; Choi, Eui‐Sung

doi:10.1016/j.procbio.2009.06.009

Cited by 15 publications

(13 citation statements)

References 12 publications

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“…S1). Thus, the Dgal80 mutant offers efficient P GAL -driven heterologous gene expression with the pronounced transcriptional enhancement of the GAL promoter not only under the aerobic condition observed in our previous study (Whang et al, 2009), but also under an anaerobic alcoholic fermentation condition. Thus, the Dgal80 mutant offers efficient P GAL -driven heterologous gene expression with the pronounced transcriptional enhancement of the GAL promoter not only under the aerobic condition observed in our previous study (Whang et al, 2009), but also under an anaerobic alcoholic fermentation condition.…”

Section: Yeast Researchsupporting

confidence: 54%

“…These results indicate that use of Dgal80 mutant as a host strain could enhance transcription of P GAL10 anaerobically. Thus, the Dgal80 mutant offers efficient P GAL -driven heterologous gene expression with the pronounced transcriptional enhancement of the GAL promoter not only under the aerobic condition observed in our previous study (Whang et al, 2009), but also under an anaerobic alcoholic fermentation condition.…”

Section: Yeast Researchsupporting

confidence: 54%

“…Firstly, efficiency of P GAL10 -driven gene expression at the anaerobic alcoholic fermentation was examined using a variant (CALB14) of lipase B from C. antarctica as a reporter (Kim et al, 2007). Wild-type S. cerevisiae 2805 (MATa pep4::HIS3 prb1-d can1 his3 ura3-52) was transformed with pGAL-MFa opt -CALB14 opt for the secreotory production of CALB14 under a control of P GAL10 (Whang et al, 2009), and then, the transformants were cultivated under aerobic and anaerobic conditions using galactose as the sole carbon source. The lipase activity of culture supernatants was determined by measuring the release of p-nitrophenol by the action of an enzyme on p-nitrophenyl palmitate (Kim et al, 2007).…”

Section: Yeast Researchmentioning

confidence: 99%

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GALpromoter-driven heterologous gene expression inSaccharomyces cerevisiaeΔ strain at anaerobic alcoholic fermentation

et al. 2013

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The removal of Gal80 protein by gene disruption turned into efficient GAL promoter-driven heterologous gene expression under anaerobic alcoholic fermentation of Saccharomyces cerevisiae. Using lipase B from Candida antarctica as a reporter, the relative strength of GAL10 promoter (P(GAL10) ) in Δgal80 mutant that does not require galactose as an inducer was compared to those of ADH1, PDC1, and PGK promoters, which have been known to work well anaerobically in actively fermenting yeast cells under high glucose concentration. P(GAL10) in the Δgal80 mutant showed 0.8-fold (ADH1), fourfold (PDC1), and 50-fold (PGK) in promoter strength.

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Section: Yeast Researchsupporting

confidence: 54%

Section: Yeast Researchsupporting

confidence: 54%

Section: Yeast Researchmentioning

confidence: 99%

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GALpromoter-driven heterologous gene expression inSaccharomyces cerevisiaeΔ strain at anaerobic alcoholic fermentation

et al. 2013

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“…For the secretory production of Bacillus endoxylanase in S. cerevisiae, the ORF of the endoxylanase gene was constructed under the control of inducible yeast GAL10 promoter and fused to the codon-optimized leader sequence of S. cerevisiae mating factor a (MFaopt) [24]. Because the endoxylanase from Bacillus sp.…”

Section: Construction Of the Expression Plasmidsmentioning

confidence: 99%

Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae

Kim¹,

Kim²,

Nam³

et al. 2013

Journal of Life Science

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The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and MFαopt s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the MFαopt s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.

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“…The Kluyveromyces marxianus NCYC2887 strain was purchased from the National Collection of Yeast Cultures (Norwich, UK). The expression vector pYEGα and the host strain Saccharomyces cerevisiae Y2805 (MATα pep::HIS3 prb-∆1.6R can1 his3-20 ura3-52) and its ∆gal80 mutant strain were as previously described [3,14,15].…”

Section: Strains and Plasmidsmentioning

confidence: 99%

Recombinant Production of an Inulinase in a Saccharomyces cerevisiae gal80 Strain

Lim

Lee²,

Sok³

et al. 2010

J. Microbiol. Biotechnol.

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The inulinase gene (INU1) from Kluyveromyces marxianus NCYC2887 was overexpressed by using the GAL10 promotor in a ∆gal80 strain of Saccharomyces cerevisiae. The inulinase gene lacking the original signal sequence was fused in-frame to a mating factor α signal sequence for secretory expression. Use of the ∆gal80 strain allowed for the galactose-free induction of inulinase expression using a glucose-only medium. Shake-flask cultivation in YPD medium produced 34.6 U/ml of the recombinant inulinase, which was approximately 13-fold higher than that produced by K. marxianus NCYC2887. It was found that the use of the ∆gal80 strain improved the expression of inulinase in the recombinant S. cerevisiae in both aerobic and anaerobic conditions by about 2.9-and 1.7fold, respectively. A 5-l fed-batch fermentation using YPD medium was performed under aerobic condition with glucose feeding, which resulted in the inulinase production of 31.7 U/ml at the OD 600 of 67. Ethanol fermentation of dried powder of Jerusalem artichoke, an inulin-rich biomass, was also performed using the recombinant S. cerevisiae expressing INU1 and K. marxianus NCYC2887. Fermentation in a 5-l scale fermentor was carried out at an aeration rate of 0.2 vvm, an agitation rate of 300 rpm, and with the pH controlled at 5.0. The temperature was maintained at 30 o C and 37 o C, respectively, for the recombinant S. cerevisiae and K. marxianus. The maximum productivities of ethanol were 59.0 and 53.5 g/l, respectively.

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Efficient, galactose-free production of Candida antarctica lipase B by GAL10 promoter in Δgal80 mutant of Saccharomyces cerevisiae

Cited by 15 publications

References 12 publications

GALpromoter-driven heterologous gene expression inSaccharomyces cerevisiaeΔ strain at anaerobic alcoholic fermentation

GALpromoter-driven heterologous gene expression inSaccharomyces cerevisiaeΔ strain at anaerobic alcoholic fermentation

Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae

Recombinant Production of an Inulinase in a Saccharomyces cerevisiae gal80 Strain

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