Specific mutation of transglutaminase gene from Streptomyces hygroscopicus H197 and characterization of microbial transglutaminase
作者: Wenjie WanDonglan HeZhijun XueZewen Zhang
作者单位: 1South-Central University for Nationalities
刊名: Journal of Biosciences, 2017, Vol.42 (4), pp.537-546
来源数据库: Springer Journal
DOI: 10.1007/s12038-017-9707-4
关键词: Enzymatic propertiesMicrobial transglutaminaseOverlapping extension PCRPurification
英文摘要: Microbial transglutaminase (MTG) gene ( mtg ) from Streptomyces hygroscopicus H197 strain was cloned by PCR and mutated by deleting a specific 84 bp fragment using overlapping extension PCR. The mutant MTG and the wild MTG genes expressed by recombinant plasmid pET32a+-mutant mtg and pET32a+-mtg, respectively, and were harvested by alternating freeze–thaw steps and purified by Ni column. The purified mutant MTG and the wild MTG exhibited 0.22 U/mg and 0.16 U/mg activity, respectively, and 0.69 U/mg and 0.54 U/mg activity, respectively, after activated by trypsin. The molecular weight of mutant MTG was estimated as 67 kDa by SDS-PAGE. Both MTGs showed optimum activity at pH 6–8 for hydroxamate formation from N-CBZ-Gln-Gly and hydroxylamine, and exhibited higher...
原始语种摘要: Microbial transglutaminase (MTG) gene ( mtg ) from Streptomyces hygroscopicus H197 strain was cloned by PCR and mutated by deleting a specific 84 bp fragment using overlapping extension PCR. The mutant MTG and the wild MTG genes expressed by recombinant plasmid pET32a+-mutant mtg and pET32a+-mtg, respectively, and were harvested by alternating freeze–thaw steps and purified by Ni column. The purified mutant MTG and the wild MTG exhibited 0.22 U/mg and 0.16 U/mg activity, respectively, and 0.69 U/mg and 0.54 U/mg activity, respectively, after activated by trypsin. The molecular weight of mutant MTG was estimated as 67 kDa by SDS-PAGE. Both MTGs showed optimum activity at pH 6–8 for hydroxamate formation from N-CBZ-Gln-Gly and hydroxylamine, and exhibited higher...
全文获取路径: Springer  (合作)
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影响因子:1.759 (2012)

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关键词翻译
关键词翻译
  • mutant 突变体
  • purified 净化的
  • mutation 变种
  • concentration 浓度
  • expressed 榨出的
  • extension 延长
  • activity 活度
  • addition 添加
  • plasmid 质粒
  • formation 建造