绍兴黄酒成品麦曲中微生物胞外酶的双向电泳技术的建立

Establishment of a Two-Dimensional Electrophoresis Protocol for Extracellular Enzymes Excreted by Microorganisms from Shaoxing Rice Wine Wheat Qu

应用宏蛋白质组学理论和思想,利用双向电泳技术分离绍兴黄酒成品麦曲中主要包含各种微生物胞外酶的可溶性总蛋白。通过研究不同等电聚焦条件、不同pH值浸提缓冲液以及不同上样量等关键因素对双向电泳结果的影响,建立了适合于绍兴黄酒成品麦曲微生物胞外酶的双向电泳条件,得到了分辨率较高,重复性较好的双向电泳图谱,为进一步通过质谱分析手段全面鉴定麦曲中微生物胞外酶的组成,明确其微生物来源奠定了基础。

In this study,the two-dimensional（2-D） electrophoresis was used to separate the total extracellular enzymes were excreted by the microorganisms of wheat qu of Shaoxing rice wine.A 2-D electrophoresis technique which was suitable for the extracellular enzymes was established through investigating the the isoelectric focusing conditions,pH values of extraction buffer,and sampling volumes and other key factors on the effects of 2-D electrophoresis..It was demonstrated that the 2-D electrophoresis profiles had high resolution and good reproducibility.The results present here provideda basis for further identification of the extracellular enzymes excreted by the microorganisms from Shaoxing rice wine wheat qu by MALDI-TOF-MS/MS analysis.