解脂耶氏酵母异源合成番茄红素的初步研究  

Metabolic Engineering Study of Heterologous Production of Lycopene in Yarrowia lipolytica

作　　者：楼佳佳[1] 韦柳静[1] 花强[1,2] 

机构地区：[1]华东理工大学生物工程学院,生物反应器工程国家重点实验室,上海200237 [2]上海生物制造技术协同创新中心,上海200237

出　　处：《华东理工大学学报（自然科学版）》2015年第3期314-320,共7页Journal of East China University of Science and Technology

基　　金：国家重点实验室专项经费(2060204);国家自然科学基金(31200025)

摘　　要：为了探究番茄红素在解脂耶氏酵母中的生物合成,利用两个解脂耶氏酵母整合表达质粒(单拷贝质粒pINA1269和多拷贝质粒pINA1312),将成团泛菌来源的番茄红素生物合成基因(crtE、crtB、crtI)克隆到解脂耶氏酵母营养缺陷型宿主菌Po1f中实现多基因共表达,并成功构建了异源合成番茄红素的解脂耶氏酵母工程菌株。在hp4d启动子和XPR2终止子的控制下,本实验共构建了8个能产番茄红素的解脂耶氏酵母工程菌株;同时筛选了其中5个代表性菌株(Po1f-1269IBE-1312、Po1f-1312E-1269IB、Po1f-1312B-1269IE、Po1f-1312I-1269EB、Po1f-1312IBE-1269)基于摇瓶培养进一步研究其生长特性和产物合成情况。结果表明,工程菌Po1f-1312E-1269IB的培养可以获得最高番茄红素产量,相对于细菌干重其产量约为0.9mg/g。The non-conventional oleaginous yeast Yarrowia lipolytica was used as a starting strain. Three lycopene biosynthetic genes （crtE, crtB, crtI） from Pantoea agglomerans were introduced and coexpressed in the auxotrophic Y. lipolytica strain Polf. In order to systematically investigate the metabolic functions of these genes in Y. lipolytica, single-copy （pINA1269） and multi-copy （pINA1312） integrative plasmids were used to regulate the expression level of the crtE, crtB and crtI genes, under the control of the hp4d promoter and an XPR2 terminator. Eight lycopene-producing recombinant Y. lipolytica strains were therefore successfully obtained, among which five strains （Polf-1269IBE-1312, Polf-1312E-1269IB, Polf-1312B-1269IE, Polf-13121-1269EB, Polf-1312IBE-1269 ） were further investigated. The highest production of about 0.9 mg of lycopene per gram of dry cell weight was achieved for the strain Polf-1312E-1269IB coexpressed with multi-copy crtE and single-copy crtB and crtI genes.

关 键 词：解脂耶氏酵母 番茄红素 异源共表达 单拷贝整合 多拷贝整合 

分 类 号：Q815[生物学—生物工程]