亚氨基糖类碳水化合物活性酶抑制剂在代谢综合征、病毒感染和癌症方面具有治疗应用。与化学合成相比，微生物亚胺糖生产具有成本、可持续性和优化的优点。在这项研究中，来自贝莱斯芽孢杆菌 MBLB0692 的 1-脱氧野尻霉素 (DNJ) 生物合成基因簇及其单个基因被克隆到谷氨酸棒杆菌 (Cg) 中。使用带有单个和簇 (TYB) 基因的纯化酶和全细胞生物催化剂对编码的转氨酶 GabT1、磷酸酶 Yktc1 和脱氢酶 GutB1 进行表征。 GabT1 的半反应表现出可变模式，且周转缓慢。 GutB1 是一种碱性脱氢酶，具有广泛的底物特异性，不依赖于二价离子，而锌依赖性磷酸酶 Yktc1 具有既依赖于 pH 又依赖于离子的底物特异性。 CgYktc1 和 CgGutB1 全细胞是可行的生物催化剂，比其酶对应物具有更广泛的底物范围。 CgTYB 细胞产生甘露糖苷酶抑制亚胺糖，对应于甘露尻霉素脱水物 (162m/z) 和脱氧甘露尻霉素 (164m/z)。甘露糖苷酶抑制剂已被发现可有效治疗孤儿疾病、癌症和病毒感染，并且重组谷氨酸棒杆菌的生物合成可优化用于工业生产和新药开发。版权所有 © 2024。由 Elsevier B.V. 出版。

The iminosugar class of carbohydrate-active enzyme inhibitors has therapeutic applications in metabolic syndrome conditions, viral infections and cancer. Compared to chemical synthesis, microbial iminosugar production has benefits of cost, sustainability and optimization. In this study, the 1-deoxynojirimycin (DNJ) biosynthetic gene cluster from Bacillus velezensis MBLB0692, and its individual genes, were cloned into Corynebacterium glutamicum (Cg). Characterizations of the encoded aminotransferase GabT1, phosphatase Yktc1, and dehydrogenase GutB1, were performed with purified enzymes and whole cell biocatalysts bearing individual and clustered (TYB) genes. GabT1 showed a variable pattern in its half-reaction with a slow turnover. GutB1 was an alkaline dehydrogenase with a broad substrate specificity and no divalent ion dependency while the zinc-dependent phosphatase Yktc1 had substrate specificity that was both pH- and ion-dependent. The CgYktc1 and CgGutB1 whole cells were viable biocatalysts with wider ranges of substrates than their enzyme counterparts. The CgTYB cells produced mannosidase-inhibiting iminosugars corresponding to mannojirimycin dehydrate (162 m/z) and deoxymannojirimycin (164 m/z). Mannosidase inhibitors have been found to be effective in treating orphan diseases, cancer and viral infections, and their biosynthesis by recombinant C. glutamicum can be optimized for industrial production and novel drug development.Copyright © 2024. Published by Elsevier B.V.