title: Reduced production of ethyl carbamate for wine fermentation by deleting CAR1 in Saccharomyces cerevisiae.
abstract: Ethyl carbamate (EC), a pluripotent carcinogen, is mainly formed by a spontaneous chemical reaction of ethanol with urea in wine. The arginine, one of the major amino acids in grape musts, is metabolized by arginase (encoded by CAR1) to ornithine and urea. To reduce the production of urea and EC, an arginase-deficient recombinant strain YZ22 (Δcarl/Δcarl) was constructed from a diploid wine yeast, WY1, by successive deletion of two CAR1 alleles to block the pathway of urea production. The RT-qPCR results indicated that the YZ22 almost did not express CAR1 gene and the specific arginase activity of strain YZ22 was 12.64 times lower than that of parent strain WY1. The fermentation results showed that the content of urea and EC in wine decreased by 77.89 and 73.78 %, respectively. Furthermore, EC was forming in a much lower speed with the lower urea during wine storage. Moreover, the two CAR1 allele deletion strain YZ22 was substantially equivalent to parental strain in terms of growth and fermentation characteristics. Our research also suggested that EC in wine originates mainly from urea that is produced by the arginine.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.|Tianjin Food Safety and Low Carbon Manufacturing Collaborative Innovation Center, Tianjin, 300457, People's Republic of China.
authors.forename: Xue-Wu
authors.initials: XW
authors.lastname: Guo
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.
authors.forename: Yuan-Zi
authors.initials: YZ
authors.lastname: Li
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: Guo
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.
authors.forename: Shi-Yong
authors.initials: SY
authors.lastname: Huang
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.|Tianjin Food Safety and Low Carbon Manufacturing Collaborative Innovation Center, Tianjin, 300457, People's Republic of China.
authors.forename: Ye-Fu
authors.initials: YF
authors.lastname: Chen
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China.|Tianjin Food Safety and Low Carbon Manufacturing Collaborative Innovation Center, Tianjin, 300457, People's Republic of China.
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Du
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, Tianjin, 300457, People's Republic of China. xdg@tust.edu.cn.|College of Bioengineering, Tianjin University of Science and Technology, No. 29, 13th Avenue, TEDA-Tianjin Economic-Technological Development Area, Tianjin, 300457, People's Republic of China. xdg@tust.edu.cn.|Tianjin Food Safety and Low Carbon Manufacturing Collaborative Innovation Center, Tianjin, 300457, People's Republic of China. xdg@tust.edu.cn.
authors.forename: Dong-Guang
authors.initials: DG
authors.lastname: Xiao
mesh_terms: D000483:Alleles; D001119:Arginase / Q000235:genetics* / Q000378:metabolism; D001120:Arginine / Q000378:metabolism; D002273:Carcinogens / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D009952:Ornithine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D014508:Urea / Q000378:metabolism; D014520:Urethane / Q000378:metabolism*; D014920:Wine / Q000032:analysis* / Q000382:microbiology*
chemical_list: D002273:Carcinogens; D029701:Saccharomyces cerevisiae Proteins; D014520:Urethane; D000431:Ethanol; D014508:Urea; D001120:Arginine; D009952:Ornithine; D001119:Arginase; C490028:CAR1 protein, S cerevisiae
keywords: CAR1; Ethyl carbamate; Saccharomyces cerevisiae; Urea; Wine
references.citation: J Biol Chem. 1973 Sep 10;248(17):6203-9
references.pmid: 4580053
references.citation: J AOAC Int. 1994 Nov-Dec;77(6):1530-6
references.pmid: 7819763
references.citation: Int J Food Microbiol. 2014 Jun 16;180:19-23
references.pmid: 24769164
references.citation: Mol Cell Biol. 1986 Nov;6(11):3954-64
references.pmid: 3025621
references.citation: DNA Seq. 1991;2(1):19-32
references.pmid: 1802034
references.citation: Appl Microbiol Biotechnol. 2007 Mar;74(4):843-50
references.pmid: 17216464
references.citation: Cancer Lett. 2003 Aug 20;198(2):139-44
references.pmid: 12957351
references.citation: Toxicol Lett. 2003 Apr 30;142(1-2):111-7
references.pmid: 12765245
references.citation: Nucleic Acids Res. 2002 Mar 15;30(6):e23
references.pmid: 11884642
references.citation: J Ind Microbiol Biotechnol. 2015 Apr;42(4):617-25
references.pmid: 25616436
references.citation: Appl Environ Microbiol. 1991 Jan;57(1):301-6
references.pmid: 2036017
references.citation: J Agric Food Chem. 2014 Apr 2;62(13):2797-802
references.pmid: 24611619
references.citation: Free Radic Biol Med. 2002 Sep 1;33(5):703-14
references.pmid: 12208357
references.citation: Food Chem Toxicol. 2005 Jan;43(1):1-19
references.pmid: 15582191
references.citation: J Agric Food Chem. 2007 Feb 7;55(3):608-13
references.pmid: 17263449
references.citation: Curr Genet. 1989 Dec;16(5-6):339-46
references.pmid: 2692852