Production of the α-glycosidase inhibitor 1-deoxynojirimycin from Bacillus species
Production of the α-glycosidase inhibitor 1-deoxynojirimycin from Bacillus species
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DOI：
10.1016/j.foodchem.2012.11.012
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发表时间：
2013-05-01
期刊：
影响因子：
8.8
通讯作者：
Miyazawa, Teruo
中科院分区：
文献类型：
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作者：
Onose, Shinji;Ikeda, Ryoichi;Miyazawa, Teruo
1-脱氧野尻霉素（DNJ）是一种有效的α-糖苷酶抑制剂，在治疗HIV、戈谢病和糖尿病方面具有治疗应用。DNJ已从天然来源（桑葚）中提取用于治疗目的;然而，DNJ成分供应有限，并且大规模获得成本高。由于某些芽孢杆菌属和链霉菌属物种的菌株据报道产生DNJ，它们可以作为高产量DNJ生产的潜在来源。在这项研究中，我们获得了证据的DNJ生产枯草芽孢杆菌DSM 704的亲水相互作用色谱-串联质谱。此外，从750种微生物的筛选中，我们鉴定了另外的大量产生DNJ的芽孢杆菌菌株（解淀粉芽孢杆菌AS 385和枯草芽孢杆菌B4）。使用枯草芽孢杆菌DSM 704和DNJ高产芽孢杆菌菌株研究各种培养条件的影响，提供了山梨醇补充对DNJ前体2-氨基-2-脱氧-D-甘露醇产量的重要性的证据，从而增加DNJ产量。山梨醇在增加DNJ产量中的作用得到了所观察到的生物合成基因gabT 1 mRNA表达增加的支持。当在补充有山梨醇的培养基中培养解淀粉芽孢杆菌AS 385时，细胞外DNJ浓度达到最大值460 mg/l培养基（相当于9.20 mg/g冻干培养基），表明该菌株可作为食品级和药品级产品的来源。这些发现不仅导致对DNJ生物合成途径的进一步理解，而且还提出了用于治疗应用的微生物大量生产DNJ的方法。(C)2012爱思唯尔有限公司保留所有权利。
1-Deoxynojirimycin (DNJ), a potent alpha-glycosidase inhibitor, has therapeutic applications in treatments of HIV, Gaucher's disease, and diabetes. DNJ has been extracted from natural sources (mulberry leaves) for therapeutic purposes; however, DNJ ingredients are in limited supply and are costly to obtain on a large scale. Since certain strains of Bacillus and Streptomyces species reportedly produce DNJ, they may serve as potential sources for high-yield DNJ production. In this study, we obtained evidence for a DNJ production in Bacillus subtilis DSM704 by hydrophilic interaction chromatography-tandem mass spectrometry. In addition, from a screen of 750 microorganisms, we identified additional Bacillus strains (Bacillus amyloliquefaciens AS385 and Bacillus subtilis B4) that produce DNJ in large quantities. Investigation of the effect of various culture conditions, using Bacillus subtilis DSM704 and the DNJ high-production Bacillus strains, provided evidence for the importance of sorbitol supplementation on the yield of the DNJ precursor, 2-amino-2-deoxy-D-mannitol, thereby increasing DNJ production. The role of sorbitol in increased DNJ production was supported by an observed increase in mRNA expression of the biosynthetic gene, gabT1 When Bacillus amyloliquefaciens AS385 was cultured in medium supplemented with sorbitol, extracellular DNJ concentration reached a maximum of 460 mg/l of medium (equivalent to 9.20 mg/g of freeze-dried medium), indicating that this strain can serve as a source for food- and drug-grade products. These findings not only lead to a further understanding of the DNJ biosynthetic pathway, but also suggest a method for microbial mass production of DNJ for therapeutic applications. (C) 2012 Elsevier Ltd. All rights reserved.