Antisense-mediated inhibition of arginase (CAR1) gene expression in Saccharomyces cerevisiae

Park, Heui-Dong; Shin, Min-Choul; Woo, Im-Sun

doi:10.1016/s1389-1723(01)80302-9

Cited by 18 publications

(13 citation statements)

References 22 publications

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Section: Methodsmentioning

confidence: 99%

“…S. cerevisiae TCY1 (a, lys2, ura3) was used for yeast transformation and acid trehalase activity assay (Park et al 2001). Plasmids YMCp31 (Park et al 2001) and pLG669z (Kovari et al 1990) were used as a plasmid vector. Yeast was grown at 30°C in YPD media (1% yeast extract, 2% Bacto-peptone, and 2% glucose) for seed culture and YNB media (0.17% yeast nitrogen base w/o amino acid and ammonium sulfate, 0.5% ammonium sulfate, 2% glucose) supplemented with 25 mg L-lysine l )1 for the selection of yeast transformants by plasmids containing the URA3 gene as a selective marker (Kaiser et al 1989).…”

Section: Methodsmentioning

confidence: 99%

“…The sequences of the primers are shown in Figure 1. A KpnI site at 5¢-end and a BamHI site at 3¢-end of the PCR products were constructed using PCR primers for the cloning into the BamHI and KpnI sites downstream of the yeast CYC1 promoter on the YMCp31 plasmid (Park et al 2001). After PCR, using the primers anti-ATH1F and anti-ATH1R shown in the Figure 1 and the yeast chromosomal DNA as a template, the PCR products were resolved in a 1% agarose gel and confirmed as containing the amplified 500 bp DNA.…”

Section: Construction Of Recombinant Plasmids Containing Antisense Atmentioning

confidence: 99%

“…After PCR, using the primers anti-ATH1F and anti-ATH1R shown in the Figure 1 and the yeast chromosomal DNA as a template, the PCR products were resolved in a 1% agarose gel and confirmed as containing the amplified 500 bp DNA. The amplified DNA, digested with BamHI and KpnI, was fused to the yeast CYC1 promoter on the plasmid YMCp31 (Park et al 2001) as shown in Figure 1 to produce a recombinant plasmid YJE12 containing antisense ATH1 DNA fused to yeast CYC1 promoter.…”

Section: Construction Of Recombinant Plasmids Containing Antisense Atmentioning

confidence: 99%

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Antisense-Mediated Inhibition of Acid Trehalase (ATH1) Gene Expression Promotes Ethanol Fermentation and Tolerance in Saccharomyces cerevisiae

Jung

Park

2005

Biotechnol Lett

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Acid trehalase gene (ATH1) expression was decreased using the antisense-RNA technique in Saccharomyces cerevisiae. The 500 bp DNA fragments containing anti-ATH1 gene between +1 and +500 were amplified using PCR and fused to yeast ADH1, CYC1 and ATH1 promoters. Yeast cells harboring the recombinant plasmids had a low activity of acid trehalase and promoted ethanol fermentation compared to the control yeast cells harboring the vector plasmid only. The recombinant yeast had a high viability with 8% (v/v) ethanol.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Construction Of Recombinant Plasmids Containing Antisense Atmentioning

confidence: 99%

Section: Construction Of Recombinant Plasmids Containing Antisense Atmentioning

confidence: 99%

See 2 more Smart Citations

Antisense-Mediated Inhibition of Acid Trehalase (ATH1) Gene Expression Promotes Ethanol Fermentation and Tolerance in Saccharomyces cerevisiae

Jung

Park

2005

Biotechnol Lett

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“…However, a high EC concentration (73.2 µg L −1 -243.9 µg L −1 ) was detected in YW (Fu et al, 2010;Wang et al, 2010;Wu et al, 2014). EC can be reduced by adding urease into fermentation mash or beverage to decompose the urea (Fujinawa et al, 1990;Ough et al, 1990;Miyagawa et al, 1999) or by inhibiting the arginase activity in yeast with antisense RNA interference or Car1 gene knockout (Parr et al, 2001). Car1 gene is deleted successfully from the sake yeast strain (Kitamoto et al, 1991), but there is no report on construction of genetically modified YW yeast to reduce EC formation.…”

Section: Introductionmentioning

confidence: 99%

Construction of car1 Deletion Mutant fromSaccharomyces cerevisiaeand Evaluation of Its Fermentation Ability

Zhang

Chen

2015

Food Biotechnology

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In order to reduce ethyl carbamate content in yellow wine (YW), a car1 deletion mutant (S car1) was constructed by SFH-PCR-mediated gene knockout and the YW fermentation process of the S car1 were analyzed. The results indicated that S car1 was successfully constructed. The stationary phase of the S car1 was at 30 h and 6 h later than that of the wild strain (24 h).The arginase activity in cultural supernatant of S car1 was not detected, while the 5.2 U activity was detected in that of the wild strain after being cultured for 18 h in YNBG. The fermentation profile of the S car1 was basically the same with that of the wild strain. The arginine (5.29 g/100 L), lysine (3.16 g/100 L) and cysteine concentrations (1.38 g/100 L) in the YW brewed with S car1 were higher than that in the YW brewed with wild strain (0.44 g/100 L,1.46 g/100 L and 0.63 g/100 L, respectively),but the ornithine concentration (0.58 g/100 L) was lower than that in the YW brewed with wild strain (1.98 g/100 L).The EC concentration in YW brewed with S car1 (5.93 µg/L) was notably lower than that in YW brewed with wild strain (40.65 µg/L).The successful construction of S car1 would provide a strain to produce YW with low concentration EC. However, the safety of the mutant should be assessed before it is used as starter.

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Enhancement of urea uptake in Chinese rice wine yeast strain N85 by the constitutive expression ofDUR3for ethyl carbamate elimination

Shen

et al. 2015

J. Inst. Brew.

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Most of the fermented alcoholic beverages, particularly Chinese rice wine, contain the potentially human carcinogenic compound ethyl carbamate (EC). As a major EC precursor in Chinese rice wine, urea in fermentations can be transported into the yeast cell by urea permease and finally metabolized by urea carboxylase and allophanate hydrolase in vivo. To eliminate EC in Chinese rice wines, the present study constructed high urea uptake yeast strains N1-D, N2-D and N-D, by introducing a strong promoter (PGK1p) into the urea permease gene (DUR3) of the industrial Chinese rice wine yeast N85, and by the restoration of the URA3 gene at the same time. With these self-cloned, high urea uptake strains, the urea and EC in the terminal Chinese rice wine samples were reduced to different extents. With two copies of overexpressed DUR3, the N-D strain could reduce the urea and the EC by 53.4 and 26.1%, respectively. No difference in fermentation characteristics was found between the engineered strains and the parental industrial yeast strain N85. These results could help to optimize the genetic manipulation strategy for EC elimination in Chinese rice wine production.

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Antisense-mediated inhibition of arginase (CAR1) gene expression in Saccharomyces cerevisiae

Cited by 18 publications

References 22 publications

Antisense-Mediated Inhibition of Acid Trehalase (ATH1) Gene Expression Promotes Ethanol Fermentation and Tolerance in Saccharomyces cerevisiae

Antisense-Mediated Inhibition of Acid Trehalase (ATH1) Gene Expression Promotes Ethanol Fermentation and Tolerance in Saccharomyces cerevisiae

Construction of car1 Deletion Mutant fromSaccharomyces cerevisiaeand Evaluation of Its Fermentation Ability

Enhancement of urea uptake in Chinese rice wine yeast strain N85 by the constitutive expression ofDUR3for ethyl carbamate elimination

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