食品科学技术学报2025，Vol.43Issue(6)：96-106,11.DOI:10.12301/spxb202500210
热袍菌α-葡聚糖磷酸化酶在枯草芽孢杆菌中的表达及应用
Expression and Application of α-Glucan Phosphorylase from Thermotoga sp.in Bacillus subtilis
摘要
Abstract
α-D-Glucose-1-phosphate(α-G1P)is a metabolic intermediate involved in a variety of metabolic reactions,and is also a potential drug,which has great development potential in the field of medicine and health supplements.It is mainly produced by α-glucan phosphorylase(αGP)using starch as substrate.In this study,α-glucan phosphorylase ThαGP from Thermotoga sp.SG1 was heterologously expressed in Bacillus subtilis.Then the types of promoters,fermentation medium formulation,temperature and time were optimized.And the application experiment of preparing α-G1P was carried out.The results showed that the enzyme activity of recombinant strain B.subtilis WS9/pHY300PLk-SpovG-amyQ-thαgp was 7.01-fold higher than that of the original strain fermentation.After optimization of shake flask fermentation conditions,the total activity of ThαGP reached 173.38 U/mL after fermentation at 33℃for 60 h in a shaking flask using medium with 18 g/L tryptone and 30 g/L yeast extract as complex nitrogen source and 15 g/L soluble starch as carbon source,which was 4.13 times that of the original fermentation.When ThαGP was combined with pullulanase and 4-α-glucosyltransferase,the conversion rate of α-G1P catalyzed by ThαGP was up to 51.62%.It was hoped that the study would provide a reference and technical foundation for the food grade fermentation preparation of ThαGP and industrial production of α-G1P.关键词
α-D-葡萄糖-1-磷酸/α-葡聚糖磷酸化酶/枯草芽孢杆菌/启动子优化/发酵优化Key words
α-D-glucose-1-phosphate/α-glucan phosphorylase/Bacillus subtilis/promoter optimization/fermentation optimization分类
轻工纺织引用本文复制引用
LIANG Xinsheng,WU Jing,XIA Wei..热袍菌α-葡聚糖磷酸化酶在枯草芽孢杆菌中的表达及应用[J].食品科学技术学报,2025,43(6):96-106,11.基金项目
国家重点研发计划项目(2022YFC2104900).National Key Research and Development Program of China(2022YFC2104900). （2022YFC2104900）
