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阿卡波糖(Acarbose)作为α-葡萄糖苷酶抑制剂类药物,因其高效、安全而被广泛应用于2型糖尿病的治疗,工业上主要采用游动放线菌(Actinoplanes spp.)发酵生产。解析阿卡波糖生物合成过程中的分子机制并进一步提升产量是目前亟待解决的问题。本研究首先考察了不同麦芽糖浓度对阿卡波糖产量的影响,发现随着麦芽糖浓度提高,阿卡波糖产量呈先上升后下降的趋势,最适浓度为50 g/L。同时,检测了不同麦芽糖浓度下阿卡波糖合成相关基因的转录水平,发现麦芽糖浓度提高显著促进转运蛋白基因acbW的转录。为了进一步考察acbW基因对阿卡波糖产量的影响,在Actinoplanes sp. SE50/110中将其缺失后,获得的突变株生长减弱,且无阿卡波糖积累,而分别加强表达acbW和acbWXY基因后,阿卡波糖产量提高了6.4%和8%。该研究表明,acbWXY编码的转运蛋白在阿卡波糖的生产过程中至关重要,且强化该外运途径可有效提高阿卡波糖产量,为菌株高产改造提供了重要依据。
Abstract：Acarbose, an α-glucosidase inhibitor, is produced in large scale by Actinoplanes spp. and used widely for the treatment of type-2 diabetes because of its high efficiency and safety. Elucidating the mechanism of acarbose biosynthesis and enhancing the yield are urgent. In this study, the effect of different maltose concentrations on acarbose production was determined, the results showed that the acarbose yield tend to increased first then decreased by increasing maltose concentrations, the optimal maltose concentration for cell growth and acarbose production is 50 g/L. Meanwhile, the transcription of acb cluster was determined with different maltose concentrations, the transcription of acbW, a gene proposed to encode the acarbose exporter was dramatically improved by increasing maltose concentrations. To further investigate the involvement of acbW in acarbose production, it was inactivated, the growth of the mutant was weakened and the accumulation of acarbose was abolished in the mutant. Moreover, the overexpression of acbW and acbWXY improved the acarbose yield by 6.4%and 8%, respectively. These results suggested that the exporter AcbWXY is necessary for acarbose production and the overexpression of acbWXY is an efficient strategy for improving acarbose production. This study provides an important reference for high-yield transformation of acarbose-producing strains.
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基本信息:
DOI：10.13417/j.gab.038.004536
中图分类号:TQ464.1;TQ920.1
引用信息:
[1]解慧欣,赵芹芹,白林泉.麦芽糖及转运蛋白对游动放线菌生产阿卡波糖的影响[J].基因组学与应用生物学,2019,38(10):4536-4544.DOI:10.13417/j.gab.038.004536.
基金信息:
国家自然科学基金(No.31470157)资助
2018-04-23
2018-04-23
2018-04-23