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酒酒球菌mleA基因重组表达质粒pYELmleA的构建及其在酿酒酵母中的表达 被引量：1

Construction of Recombinant Expression Plasmid pYELmleA of mleA Gene and Expression in Saccharomyces cerevisiae
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摘要 苹果酸-乳酸酶是进行MLF的功能酶.笔者进行酒酒球菌SD-2a的苹果酸-乳酸酶基因重组表达质粒的构建,利用来自重组质粒pLmleA的mleA基因,以PGK1强启动子和ADH1终止子为调控元件,以大肠杆菌-酵母菌穿梭质粒YEp352为载体,构建了重组表达质粒pYELmleA,并转化酿酒酵母(Saccharomyces cerevisiae)YS58.酵母转化子用含有亮氨酸、组氨酸和色氨酸的YNB平板筛选鉴定.SDS-PAGE检测表明获得的转化子表达了约60KD的目标蛋白,斑点杂交检测表明目的基因mleA转化到受体菌中.获得的转化子在添加了L-苹果酸的培养基中培养4d;取培养液上清用HPLC检测L-苹果酸及L-乳酸含量,采用t检验进行差异显著性分析,结果表明nleA基因进行了功能性的表达,将L-苹果酸转化成L-乳酸,L-苹果酸和L-乳酸含量分别与对照差异极显著和显著,L-乳酸的生成量为1002-1106mg/L,苹果酸的相对降低率为19.16-22.34%.
作者 刘延琳 蒋思欣 何秀萍 李华 张博润
机构地区 西北农林科技大学葡萄酒学院 中国科学院微生物研究所
出处 《中国农学通报》 CSCD 2005年第4期48-52,共5页 Chinese Agricultural Science Bulletin
基金 陕西省自然科学基金，西北农林科技大学校科研和教改项目
关键词 重组表达质粒 LEA基因 酿酒酵母 构建 球菌 L-苹果酸 乳酸含量 L-乳酸 显著性分析 转化子 重组质粒 调控元件 强启动子 穿梭质粒 大肠杆菌 筛选鉴定 PAGE 目的基因 杂交检测 HPLC MLF 酶基因 终止子 酵母菌 亮氨酸 色氨酸
分类号 S631.201 [农业科学—蔬菜学] Q78 [生物学—分子生物学]
  • 相关文献

参考文献16

  • 1Pascal R G, Denis D, Bernard D, et al. Handbook of Enology[M]. London: John Wiley & Sons Ltd. 2000.
  • 2Labarre C, Guzzo J, Cavin J F, Divies C. Cloning and characterization of the genes encoding malolactic enzyme and the malate permease of Leuconostoc oenos[J]. Appl Environ Microbiol. 1996,62(4): 1274～1282.
  • 3Dequin S. The potential of genetic engineering for improving wine-making and baking yeasts[J]. Appl Microbiol Biotechnol,2001, 56:577～588.
  • 4Ansanay V, Dequin S, Blondin B, Barre P. Cloning, sequence and expression of the gene encoding the malolactic enzyme from Lactococcus lactis[J]. FEBS Lett., 1993, 332(1-2):74～80.
  • 5Denayrolles M, Aigle M, Lonvaud-Funel A. Functional expression in Saccharomyce scerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme[J]. FEMS Microbiol Lett., 1995,125(1):37～43.
  • 6刘延琳,蒋思欣,李华,张博润.苹果酸-乳酸发酵相关基因克隆及其在酵母中的表达[J].中国生物工程杂志,2003,23(5):27-30. 被引量：5
  • 7刘延琳,李华,蒋思欣,张博润.苹果酸-乳酸发酵的相关酶和基因的研究进展[J].微生物学通报,2003,30(4):103-107. 被引量：11
  • 8Ansanay V, Dequin S, Camarasa C, et al. Malolactic fermentation by engineered Saccharomyces cerevisiae as compared with engineered Schizosaccharomyces pombe[J]. Yeast, 1996, 12(3):215～225.
  • 9Bony M, Bidart F, Camarasa C, et al. Metabolic analysis of S. cerevisiae strains engineered for malolactic fermentation [J]. FEB S Lett., 1997,410(2-3):452～6.
  • 10Volschenk H, Viljoen M, Grobler J, et al. Engineering pathways for malate degradation in Saccharomyces cerevisiae [J]. Nat Biotechnol.1997,15(3):253～7.

二级参考文献40

  • 1[2]Labarre C, Guzzo J, Cavin J F, et al. Cloning and characterization of the genes encoding malolactic enzyme and the malate permease of Leuconostoc oenos [J]. Appl Environ Microbiol, 1996,62(4) : 1274-1282.
  • 2[3]Dequin S. The potential of genetic engineering for improving wine-makingand baking yeasts[J]. Appl Microbiol Biotechnol,2001, 56: 577-588.
  • 3[4]Ansanay V, Dequin S, Blondin B, et al. Cloning, sequence and expression of the gene encoding the malolactic enzyme from Lactococcus lactis[J]. FEBS Lett, 1993, 11:332 (1-2): 74-80.
  • 4[5]Denayrolles M, Aigle M, Lonvaud-Funel A. Functional expression in Saccharomyce scerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme[J]. FEMS Microbiol Lett, 1995 Jan 1;125(1): 37-43.
  • 5[9][美]Sambrook J,Fritsch E F and ManiatisT著,金冬雁,黎孟枫泽.分子克隆实验指南[M](第2版).北京:科学出版社.1998:924-936.
  • 6[10]Zavaleta A J, Martinez-Murcia A J and Rodriguez-Valera F, et al. Intraspecific genetic diversity of Oenococcus oeni as derived from DNA fingerprinting and sequence analyses[J]. Appl Environ Microbiol, 1997, 634: 1261-1267.
  • 7Lonvaud-Funel A. Lactic acid bacteria in the quality improvement and depreciation of wine. Antonie Van Leeuwenhoek, 1999,76( 1-4) :317 ~ 331.
  • 8Maieas S. The use of alternative technologies to develop malolactic fermentation in wine. Appl Microbiol Biotechnol,2001,56(1-2) :35~ 39.
  • 9Dequin S. The potential of genetic engineering for improving wine-making and baking yeasts. Appl Microbiol Biotechnol,2001,56:577- 588.
  • 10Williams S A,Hodges R A,Strike T L, et al. Cloning the gene for the malolactic fermentation of wine from Lactobacillus delbrueckii in Escherichia coli and yeasts. Appl Environ Microbiol, 1984,47:288 ~ 293.

共引文献24

同被引文献12

  • 1刘延琳,蒋思欣,李华,何秀萍,张博润.酒酒球菌苹果酸-乳酸酶基因的测序及分析[J].微生物学杂志,2004,24(6):29-30. 被引量：5
  • 2李华,刘延琳.酒类酒球菌mle基因座的克隆和测序[J].农业生物技术学报,2005,13(3):400-401. 被引量：2
  • 3Adams A,Gottschling D E and Kaiser C A.Methods in Yeast Genetics:A Cold Spring Harbor Laboratory Course Manual (J).New York:Cold Spring Harbor Laboratory Press,1997.99～102
  • 4Ansanay V,Dequin S,Blondin B and Barre P.Cloning,sequence and expression of the gene encoding the malolactic enzyme from Lactococcus lactis(J).FEBS Letters,1993,332(1～2):74～80
  • 5Ausubel F M,Kingston R E and Seidman J G.Short Protocols in Molecular Biology(M).Beijing:Science Press,1999.
  • 6Bony M,Bidart F,Camarasa C,Ansanay V,Dulau L,Barre P and Dequin S.Metabolic analysis of S.cerevisiae strains engineered for malolactic fermentation (J).FEBS Letters,1997,30,410 (2～3):452～456
  • 7Denayrolles M,Aigle M and Lonvaud-Funel A.Functional expression in Saccharomyce scerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme(J).FEMS Microbiology Letters,1995,125(1):37～43
  • 8Labarre C,Guzzo J,Cavin J F and Divies C.Cloning and characterization of the genes encoding malolactic enzyme and the malate permease of Leuconostoc oenos(J).Applied and Environmental Microbiology,1996,62 (4):1274～1282
  • 9Ling D W(凌代文)and Dong X Z(东秀珠).Lactic Acid Bacterium Classification & Identification and experimental Methods (M).Beijing:China Light Industry Publishing House,1999.
  • 10Sambrook J,Fritsch E F and Maniatia T.Molecular Cloning:A Laboratory Manual (M).3nd edition.Beijing:Science Press,2002.

引证文献1

二级引证文献8

中国农学通报
2005年 第4期

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