Transglutaminases (TGases) are a class of transferases widely used in the food and biotechnology industries. In this work, we describe the production of recombinant Bacillus amyloliquefaciens TGase in Escherichia coli, obtaining the protein in its soluble and active form. In order to reduce TGase activity inside host cells and consequently its toxicity, we constructed a bicistronic plasmid containing the B. amyloliquefaciens TGase gene fused to the inhibitory Streptomyces caniferus prodomain. To make the enzyme active and avoid the need of prodomain removal in vitro, we also cloned the 3C protease gene into the same plasmid. After a fast single-step purification protocol, we obtained a partially purified recombinant TGase with 37 mU/mg protein activity, that crosslinked bovine serum albumin (BSA). This is the first report on the expression of B. amyloliquefaciens TGase in E. coli in its mature and active form.

中文翻译：

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使用双顺反子载体构建在大肠杆菌中克隆和表达解淀粉芽孢杆菌转谷氨酰胺酶基因

转谷氨酰胺酶 (TGase) 是一类广泛用于食品和生物技术行业的转移酶。在这项工作中，我们描述了在大肠杆菌中生产重组解淀粉芽孢杆菌 TGase，获得可溶性和活性形式的蛋白质。为了降低宿主细胞内的 TGase 活性及其毒性，我们构建了一个双顺反子质粒，其中包含与抑制性 Streptomyces caniferus 前域融合的解淀粉芽孢杆菌 TGase 基因。为了使酶具有活性并避免体外去除前域的需要，我们还将 3C 蛋白酶基因克隆到同一质粒中。经过快速的单步纯化方案，我们获得了部分纯化的重组 TGase，其蛋白活性为 37 mU/mg，即交联牛血清白蛋白 (BSA)。这是关于 B 表达的第一份报告。