摘要
采用常温常压等离子体(ARTP)诱变方法,以耐20 g/L的苯乙酮毒性和酮基还原酶酶活作为筛选标记,对乳酸克鲁维酵母(Kluyveromyces lactis)进行选育,得到酮基还原酶酶活2.72 U/m L的突变菌株KL5,并研究了该菌株在不同碳源、氮源中的生长特性及对底物苯乙酮转化能力。结果表明,蔗糖、酵母浸粉分别是突变株的最佳碳源和氮源;在一次添加苯乙酮20 g/L底物,蔗糖22.0 g/L,酵母浸粉10.0 g/L,磷酸二氢钾3.0 g/L,七水合硫酸镁1 g/L的催化条件下,28℃、200 r/min转化48 h,诱变菌株KL5对苯乙酮转化率可达到91.8%,比出发菌株提高2.5倍。该菌株具有广阔的生物催化应用前景。
Using resistance 20 g/L acetophenone toxicity ketoreductase activity as selection marker, Kluyveromyces lactis was screened by atomspher- ic room temperature plasma （ART P） mutation method, and the mutant strain KL5 with 2.72 U/ml ketoreductase activity was obtained. The growth characteristics in different carbon sources and nitrogen sources and the biotransformation ability to acetophenone of the mutant strain KL5 were fur-ther investigated. The results showed that the sucrose and yeast extract powder were optimal carbon source and nitrogen source of mutant strain, respectively. Under the catalysis conditions of substrate acetophenone 20 g/L, sucrose 22.0 g/L, yeast extract powder 10.0 g/L, KH2 PO4 3.0 g/L and MgSO·7H2O 1 g/L, at the conversion condition of 28 ℃, 200 r/min for 48 h, the conversion rate of mutant strain KL5 to acetophenone was up to 91.8%, which was 2.5 times higher than that of the starting strain. The strain had a wide application prospect in biocatalysis field.
作者
曹纲
刘云
郭金玲
吕育财
余华顺
龚大春
CAO Gang;LIU Yun;GUO Jinling;LYU Yucai;YU Huashun;GONG Dachun(Biocatalysis Key Laboratory,China Three Gorges University,Yichang 443002, China;Department of Enzyme Production,An g e l Yeast Co.,Ltd.,Yichang 443002, China)
出处
《中国酿造》
CAS
北大核心
2018年第5期42-47,共6页
China Brewing
基金
国家自然基金项目(21776162)
宜昌市科技攻关项目(A2015-001)
