为获得适合粉丝制作工艺的普鲁兰酶,研究了嗜热脂肪土芽孢杆菌(Geobacillus stearothermophilus)普鲁兰酶基因GsP在不同枯草芽孢杆菌(Bacillus subtilis)宿主菌中的表达及重组酶的应用效果。分别以淀粉酶基因缺失的枯草芽孢杆菌1A717和蛋白酶基因缺失的菌株WB600为宿主菌,构建了表达GsP的重组菌,酶活检测和SDS-PAGE分析表明,两种重组菌产生的普鲁兰酶均为胞外酶,分别命名为GsP1a和GsPwb。两种重组酶分别用于粉丝制作,以GsP1a处理的芡糊制作的粉丝成品断条率接近于0,由GsPwb处理的芡糊制作的粉丝断条率高于前者,这可能与宿主菌WB600在发酵过程中产生的α-淀粉酶有关。该研究表明,中性普鲁兰酶是粉丝制作中明矾的理想替代物,酶液中α-淀粉酶活性的去除有利于其应用性能的提升。
In order to obtain a recombinant pullulanase suitable for vermicelli production, the pullulanase encoding gene GsP from Geobacillus stearothermophilus was secretorily expressed in Bacillus subtilis 1A717 and WB600. The recombinant pullulanase expressed in B. subtilis 1A717 and B. subtilis WB600 were GsP1a and GsPwb, respectively. Cooking experiments showed that the broken rate of vermicelli made from GsP1a-treated paste was almost 0, while the broken rate of vermicelli made from GsPwb-treated paste was notably higher due to starch degradation by α-amylase produced by host strain WB600. This study shows that neutrophilic pullulanase is an ideal alum substitute in vermicelli production, and inactivating α-amylase is beneficial to improve the quality of the product.
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