Washizu K, Ando K, Koikeda S, Hirose S, Matsuura A, Takagi H, Motoki M, Takeuchi K
Tsukuba Research Laboratories, Amano Pharmaceutical Co., Ltd., Ibaraki, Japan.
Biosci Biotechnol Biochem. 1994 Jan;58(1):82-7. doi: 10.1271/bbb.58.82.
The microbial transglutaminase (TGase)-producing strains 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.
产微生物转谷氨酰胺酶（TGase）的菌株S-8112[《农业生物化学》，53，2613 - 2617(1989)]被鉴定为茂原链霉菌的一个变种。我们使用根据TGase氨基酸序列合成的寡核苷酸，通过聚合酶链反应（PCR）扩增了一个部分基因片段，并以PCR扩增片段为探针克隆了TGase基因。该基因编码一个由406个氨基酸残基组成的TGase前体，其中包括75个氨基酸残基的前原区域和331个氨基酸残基的成熟区域。我们在酪氨酸酶启动子的控制下，在变铅青链霉菌中表达了TGase基因，并发现了一种有活性的成熟重组酶，这表明该基因产物得到了加工处理。
