Improving the enzymatic activity of<scp>l</scp>-amino acid α-ligase for imidazole dipeptide production by site-directed mutagenesis
Improving the enzymatic activity of<scp>l</scp>-amino acid α-ligase for imidazole dipeptide production by site-directed mutagenesis
复制标题
通过定点诱变提高<scp>l</scp>-氨基酸α-连接酶生产咪唑二肽的酶活性
DOI：
10.1093/bbb/zbac213
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kuramoto Ayumu
中科院分区：
文献类型：
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作者：
Kino Kuniki;Komabayashi Takuma;Hashida Ayaka;Kuramoto Ayumu
咪唑二肽，含组氨酸的二肽，包括动物肌肉中的肌肽（β-丙氨酰-l-组氨酸）、鹅丝氨酸（β-丙氨酰-3-甲基-l-组氨酸）、鲸氨酸（β-丙氨酰-1-甲基-l-组氨酸）等，具有显着的抗氧化、抗疲劳等生理功能。它们是通过从天然原料（包括鸡肉和鱼肉）中提取而获得的。然而，使用天然原料存在稳定供应和大规模生产的限制。l-氨基酸α-连接酶（Lal）通过与5'-三磷酸腺苷（ATP）水解反应催化未保护的l-氨基酸形成各种二肽。在这项研究中，Lal 的定点诱变被用来建立一种通过酶法生产咪唑二肽的有效方法。与野生型 Lal 相比，我们显着提高了底物氨基酸的转化率。
Imidazole dipeptides, histidine-containing dipeptides, including carnosine (β-alanyl-l-histidine), anserine (β-alanyl-3-methyl-l-histidine), and balenine (β-alanyl-1-methyl-l-histidine) in animal muscles have physiological functions, such as significant antioxidant and antifatigue effects. They are obtained by extraction from natural raw materials, including chicken and fish meat. However, using natural raw materials entails stable supply and mass production limitations.l-amino acid α-ligase (Lal) catalyzes the formation of various dipeptides from unprotectedl-amino acids by conjugating with adenosine 5'-triphosphate (ATP) hydrolysis reaction. In this study, site-directed mutagenesis of Lal was applied to establish an efficient method for producing imidazole dipeptides by the enzymatic process. We significantly improved the conversion rate from substrate amino acids compared with wild-type Lal.