MOLECULAR-CLONING OF THE GENE FOR MICROBIAL TRANSGLUTAMINASE FROM STREPTOVERTICILLIUM AND ITS EXPRESSION IN STREPTOMYCES-LIVIDANS
MOLECULAR-CLONING OF THE GENE FOR MICROBIAL TRANSGLUTAMINASE FROM STREPTOVERTICILLIUM AND ITS EXPRESSION IN STREPTOMYCES-LIVIDANS
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DOI：
10.1271/bbb.58.82
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发表时间：
1994-01-01
影响因子：
1.6
通讯作者：
TAKEUCHI, K
中科院分区：
文献类型：
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作者：
WASHIZU, K;ANDO, K;TAKEUCHI, K
微生物转氨酶（TGase）生产菌株S-8112 [Agric. Biol. Chem.，53，2613-2617（1989）]被鉴定为茂原轮枝链霉菌的变体。我们使用从TGase的氨基酸序列合成的寡核苷酸通过聚合酶链反应（PCR）扩增部分基因片段，并使用PCR扩增片段作为探针克隆TGase的基因。该基因编码一个由406个氨基酸残基组成的TGase前体，包括75个氨基酸残基的前体区和331个氨基酸残基的成熟区。我们在变铅青链霉菌中表达了酪氨酸酶基因，并发现了一个有活性的成熟的重组酶，表明基因产物的加工。
The microbial transglutaminase (TGase)-producing strain S-8112 [Agric. Biol. Chem., 53, 2613-2617 (1989)] was identified as a variant of Streptoverticillium mobaraense. We amplified a partial gene fragment by polymerase chain reaction (PCR) using oligonucleotides synthesized from the amino acid sequence of TGase, and cloned the gene for TGase using the PCR amplified fragment as a probe. The gene encoded a precursor of TGase consisting of 406 amino acid residues, which comprised the prepro region of 75 amino acid residues and the mature region of 331 amino acid residues. We expressed the TGase gene in Streptomyces lividans under a tyrosinase promoter, and found an active and mature recombinant enzyme, indicating the processing of the gene product.