School of Food Science and Technology, Jiangnan University, Wuxi 214122, PR China.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, PR China; School of Food Science and Technology, Jiangnan University, Wuxi 214122, PR China; Collaborative Innovation Center for Food Safety and Quality Control, Jiangnan University, Wuxi 214122, PR China.
Food Res Int. 2021 Dec;150(Pt A):110735. doi: 10.1016/j.foodres.2021.110735. Epub 2021 Sep 30.
Maltopentaose-forming amylases are promising enzymes for their ability to hydrolyze starch and produce functional maltooligosaccharides. Two maltopentaose-forming amylase genes from Bacillus megaterium (BmMFA) and Saccharophagus degradans (SdMFA) were expressed heterologously and their characteristics were analyzed. BmMFA has substantial thermostability and SdMFA owns superior product specificity. The carbohydrate-binding module of SdMFA was fused with BmMFA and the fused protein showed ideal enzymatic properties and displayed potential for industrial production of maltopentaose. Under the optimized conditions, the final product containing 47.41% maltopentaose was obtained with a conversion rate of 92.67% from starch. This study provides a novel strategy for the directed modification of MFAses through protein fusion approach.
麦芽糖五糖形成淀粉酶因其能够水解淀粉并产生功能性麦芽寡糖而备受关注。本研究从巨大芽孢杆菌（Bacillus megaterium）和嗜热解淀粉沙雷氏菌（Saccharophagus degradans）中表达了两种麦芽糖五糖形成淀粉酶基因，并分析了它们的特性。BmMFA 具有很强的热稳定性，而 SdMFA 则具有优异的产物特异性。将 SdMFA 的碳水化合物结合模块与 BmMFA 融合，融合蛋白表现出理想的酶学特性，并显示出在工业生产麦芽糖五糖方面的潜力。在优化条件下，从淀粉获得的最终产物中含有 47.41%的麦芽糖五糖，转化率为 92.67%。本研究通过蛋白融合方法为 MFAses 的定向修饰提供了一种新策略。
