酿酒酵母中高效积累β-胡萝卜素的代谢途径构建  

Metabolic Pathway Construction ofβ-Carotene Accumulation in Saccharomyces cerevisiae

作　　者：王思懿 张悦 王燕燕[1] 于放[1] WANG Si-yi;ZHANG Yue;WANG Yan-yan;YU Fang(Coll.of Bio-Engin.,Dalian Polytech.Uni.,Dalian 116034)

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034

出　　处：《微生物学杂志》2023年第4期10-17,共8页Journal of Microbiology

基　　金：辽宁省教育厅科学研究项目(J2020098,J2020042);国家自然科学基金项目(31970324);大连市科技创新基金项目(2021JJ13SN76)。

摘　　要：β-胡萝卜素是一种橘黄色的脂溶性色素,属于类胡萝卜素家族,基于其丰富的生物学活性,在食品、医药、化妆品等领域具有广泛的应用。国内外对β-胡萝卜素市场需求量巨大,特别是天然来源的β-胡萝卜素具有广阔的市场前景,建立适合天然β-胡萝卜素生产的微生物细胞工厂显得尤为重要。本研究利用基因工程手段克隆了β-胡萝卜素生物合成途径中的三个基因,分别是来源于酿酒酵母(Saccharomyces cerevisiae)的香叶基香叶基焦磷酸合成酶基因(ScBTS1)、红法夫酵母(Xanthophyllomyces dendrorhous)的八氢番茄红素脱氢酶基因(XdCrtI)以及同时具有八氢番茄红素合成酶和番茄红素环化酶的双功能酶基因(XdCrtYB),构建组成型表达载体pYES2-Kan-CrtI-CrtYB-BTS1,并转化至酿酒酵母MKP-o中,最终筛选获得能够生产β-胡萝卜素的酿酒酵母基因工程菌株。结果显示,此工程菌摇瓶产量达到14.53 mg/g细胞干重(DCW)且发酵周期短,不仅可以稳定高效地积累β-胡萝卜素,同时发酵过程中无需添加诱导剂,有利于节约生产成本。因此,可以将其作为一株具有较高β-胡萝卜素生产能力的底盘菌株,为后续建立稳定高产β-胡萝卜素的微生物细胞工厂提供参考。β-carotene is a kind of orange lipid-soluble pigment,belonging to the carotenoid family.Based on its rich biological activity,β-carotene has been widely used in food,medicine,cosmetics and other fields.There is a huge market demand forβ-carotene both at home and abroad,especially the natural source ofβ-carotene has a broad market prospect.It is particularly important to establish a microbial cell factory suitable for the production of naturalβ-carotene.In this study,three genes in theβ-carotene biosynthesis pathway were cloned by genetic engineering methods.They are the geranylgeranyl diphosphate synthase gene(ScBTS1),phytoene desaturase gene(XdCrtI)and have both phytoene synthase and lycopene cyclase genes(XdCrtYB)from Saccharomyces cerevisiae and Xanthophyllomyces dendrorhous,respectively.The constitutive expression vector pYES2-Kan-CrtI-CrtYB-BTS1 was constructed and transformed into Saccharomyces cerevisiae MKP-o.Finally,a genetically engineered strain of S.cerevisiae capable of producingβ-carotene was obtained,and the yield of the shake flask reached 14.53 mg/g dry cell weight(DCW).The engineered strain has food safety and short fermentation period,which can not only accumulateβ-carotene stably and efficiently,but also save production cost without adding inducers in the fermentation process.Therefore,it can be used as a chassis strain with highβ-carotene production capacity,laying the foundation for the subsequent establishment of a microbial cell factory with stable and highβ-carotene production.

关 键 词：Β-胡萝卜素 酿酒酵母 红法夫酵母 组成型表达 代谢途径构建 萜类合成 

分 类 号：Q935[生物学—微生物学]