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首页> 外文期刊>Applied Microbiology and Biotechnology >Novel glucoamylase-type enzymes from Thermoactinomyces vulgaris and Methanococcus jannaschii whose genes are found in the flanking region of the α-amylase genes
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Novel glucoamylase-type enzymes from Thermoactinomyces vulgaris and Methanococcus jannaschii whose genes are found in the flanking region of the α-amylase genes

机译：来自寻常嗜热放线菌和詹氏甲烷球菌的新型葡糖淀粉酶型酶，其基因位于α-淀粉酶基因的侧翼区域

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摘要

A region downstream of the gene for pullulan-hydrolyzing α-amylase, TVA II, of Thermoactinomyces vulgaris R-47 was sequenced, and an open reading frame encoding an enzyme homologous to glucoamylase was found. The nucleotide sequence of this enzyme, designated TGA, consists of 1,953 base pairs corresponding to a protein of 651 amino acid residues. The TGA gene was subcloned and expressed in Escherichia coli. Enzymatic analyses showed that, like other glucoamylases, TGA produced β-D-glucose from its substrate. However, TGA hydrolyzed maltooligosaccharides such as maltotetraose and maltose more efficiently than starch, while fungal glucoamylases preferred starch to maltooligosaccharides. The primary structure of TGA resembled a putative glucoamylase from the hyperthermophilic archaeon Methanococcus jannaschii (MGA), while homologies between TGA and the fungal glucoamylases were low. The enzymatic properties of recombinant MGA produced in E. coli cells were similar to those of TGA. These findings indicate that TGA and MGA are novel glucoamylase-type enzymes with oligosaccaharide-metabolizing activity.
机译：对寻常型嗜热放线菌R-47的支链淀粉水解α-淀粉酶TVA II的基因的下游区域进行测序，发现了编码与葡糖淀粉酶同源的酶的开放阅读框。该酶的核苷酸序列称为TGA，由1,953个碱基对组成，对应于651个氨基酸残基的蛋白质。将TGA基因亚克隆并在大肠杆菌中表达。酶促分析表明，TGA与其他葡糖淀粉酶一样，从其底物产生β-D-葡萄糖。但是，TGA水解麦芽低聚糖（例如麦芽四糖和麦芽糖）比淀粉更有效，而真菌葡糖淀粉酶比淀粉低聚麦芽糖更优选淀粉。 TGA的主要结构类似于来自超嗜热古生詹氏甲烷球菌（MGA）的推定葡糖淀粉酶，而TGA与真菌葡糖淀粉酶之间的同源性较低。在大肠杆菌细胞中产生的重组MGA的酶学性质与TGA相似。这些发现表明，TGA和MGA是具有葡糖低聚糖代谢活性的新型葡糖淀粉酶型酶。

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  • 来源
    《Applied Microbiology and Biotechnology》 |2001年第4期|465-473|共9页
  • 作者

    R. Uotsu-Tomita; T. Tonozuka; H. Sakai; Y. Sakano;

  • 作者单位

    Department of Applied Biological Science Tokyo University of Agriculture and Technology 3-5-8 Saiwai-cho Fuchu Tokyo 183-8509 Japan;

    Department of Applied Biological Science Tokyo University of Agriculture and Technology 3-5-8 Saiwai-cho Fuchu Tokyo 183-8509 Japan;

    School of Food and Nutritional Science University of Shizuoka Shizuoka 422-8526 Japan;

    Department of Applied Biological Science Tokyo University of Agriculture and Technology 3-5-8 Saiwai-cho Fuchu Tokyo 183-8509 Japan;

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