菊粉酶基因克隆表达与油脂的组分分析  

Cloning and expression of inulinase gene and analysis of oil components

作　　者：赵春海 王志鹏 池振明[3] 

机构地区：[1]滨州职业学院生物工程学院,山东滨州256603 [2]中国水产科学研究院黄海水产研究所,山东青岛266071 [3]中国海洋大学海洋生命学院,山东青岛266000

出　　处：《中国酿造》2017年第12期115-119,共5页China Brewing

基　　金：山东省中青年科学家科研奖励基金(博士基金:BS2012SW001);山东省高等学校科技计划项目(J14LE58;J17KA122);国家自然科学基金(31500032)

摘　　要：为了实现解脂亚罗威亚酵母(Yarrowia lipolytica)直接利用菊粉进行油脂生产,将外切菊粉酶基因INU1与表达质粒p INA1317连接,在解脂亚罗威亚酵母(Y.lipolytica)ACA-DC尿嘧啶缺陷突变菌株中表达。以尿嘧啶缺陷型筛选作为筛选标记获得转化子C37,经过培养菊粉酶酶活达到37.15 U/mL。在2 L发酵罐中,转化子C37以菊粉为底物进行发酵,油脂产量和细胞干质量分别为49%和14 g/L。脂肪酸分析结果显示棕榈酸、硬脂酸和油酸总和占总脂肪酸的92%以上,其中油酸含量高达59%,表明通过菊粉酶基因在解脂亚罗威亚酵母中的表达,实现了以菊粉为底物一步发酵产单细胞油脂。In order to realize oil production directly from inulin by Yarrowia lipolytica, exoinulinase gene INU1 was connected with expression plasmid pINA1317 and expressed in mutant strain Y. lipolytica ACA-DC with uracil deficiency. With uracil deficiency as selection marker, transformant C37 was obtained and the inulase activity was up to 37.15 U/ml by culture. In 2 L fermenter, transformant C37 was fermented with inulin as substrate, the oil yield and cell dry mass were 49% and 14 g/L, respectively. The results of fatty acids analysis showed that the total contents of palmitic acid, stearic acid and oleic acid were more than 92% of total fatty acids, and the oleic acid content was up to 59%, which indicated that the one-step fermentation was realized to produce single cell oil with inulin through the expression of the inulinase gene in Y. lipolytica.

关 键 词：解脂亚罗威亚酵母:菊粉酶 单细胞油脂 基因表达 

分 类 号：Q815[生物学—生物工程]