萃取破碎法提取酵母醇脱氢酶的研究  

Simultaneous Cell Disruption and Aqueous Two-phase Extraction for Separation of Alcohol Dehydrogenase from Yeast

作　　者：修志龙[1] 苏志国[1] 

机构地区：[1]大连理工大学生物化学教研室

出　　处：《生物工程学报》1993年第4期342-347,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金会青年基金

摘　　要：将双水相萃取同细胞破碎结合在一起，以水平搅拌式珠磨机为设备，以聚乙二醇和硫酸铵为杨相体系，从酿酒酵母中以边破碎边萃取的方式提取醇脱氢酶。节省了萃取设备和时间，并利用双水相对蛋白质的保护作用提高了醇脱氢酶的活性。经过萃取破碎的酵母匀浆离心分相后，细胞碎片滞留在下相，９０％以上的醇脱氢酶分配在上相，分配系数大于１０，纯化倍数大于２。Aqueous two-phase extraction and cell disruption were combined and performed simultaneously in a horizontal stirred bead mill (Dyno-mill) for the separation of alcohol dehydrogenase from yeast Saccharomyces cerevisiae. Polyethylene glycol (PEG) 600 and ammonium sulphate were used as the phase forming system. The advantages of the novel process included saving additional extractor and processing time, and increasing the enzyme activity due to the protective effect of the aqueous two-phase during cell disruption. The resulting homogenate was directly centrifuged for phase separation. The cell debris was retained in the bottom phase. More than 90% of alcohol dehydrogenase was partitioned into the top phase with the partition coefficient greater than 10 and the purification factor greater than 2.

关 键 词：酵母 醇脱氢酶 提取 双水相萃取 

分 类 号：Q81[生物学—生物工程]