过表达RkCrtYB基因对红冬孢酵母类胡萝卜素合成的影响  

Effects of RkCrtYB overexpression on carotenoid production in Rhodosporidium kratochvilovae

作　　者：范晶蝶 刘韬 陈波 邱婧雯 陈媛[1] 张琦[1] FAN Jingdie;LIU Tao;CHEN Bo;QIU Jingwen;CHEN Yuan;ZHANG Qi(College of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500,Yunnan,China)

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《云南大学学报(自然科学版)》2025年第2期373-381,共9页Journal of Yunnan University(Natural Sciences Edition)

基　　金：国家自然科学基金(31660454,31960466);云南省教育厅科学研究基金(202350145)。

摘　　要：八氢番茄红素合成酶/番茄红素环化酶(CrtYB)是合成类胡萝卜素的重要酶.研究克隆了红冬孢酵母(Rhodosporidium kratochvilovae)YM25235菌株中的CrtYB基因,并命名为RkCrtYB.RkCrtYB基因全长1785 bp,编码594个氨基酸.通过对RkCrtYB基因编码的氨基酸序列进行分析,发现其与已知的CrtYB具有高度相似性.进一步的功能验证分析表明,过表达RkCrtYB基因使YM25235菌株中的类胡萝卜素质量比提高了约35.98%,同时油脂质量比也提高了约17.32%.这种现象可能是由于类胡萝卜素合成的增加需要合成更多的脂滴以储存,从而促进油脂的积累,基因转录水平的分析结果也支持了这一推测.所有结果表明,YM25235菌株中过表达RkCrtYB基因能够显著提高类胡萝卜素的合成和油脂的积累,为通过代谢工程手段开发高产类胡萝卜素的产油菌株提供了新的思路和参考.Lycopene cyclase/β-hydroxy-lycopene synthase(CrtYB)is a key enzyme involved in carotenoidbiosynthesis.In this study,we cloned the CrtYB gene from the Rhodosporidium kratochvilovae YM25235 strainand designated it RkCrtYB.The full-length RkCrtYB gene is 1785 bp and encodes 594 amino acids.Sequenceanalysis of the RkCrtYB gene revealed a high degree of similarity to known CrtYB sequences.Functionalvalidation further showed that overexpression of RkCrtYB in YM25235 resulted in a 35.98%increase in carotenoidcontent,while the lipid content also increased by approximately 17.32%.This phenomenon is likely due to theincreased synthesis of carotenoids,which requires the accumulation of additional lipid droplets for storage,therebypromoting lipid accumulation.Gene expression analysis supported this hypothesis.Collectively,our findingsdemonstrate that overexpression of RkCrtYB in YM25235 significantly enhances both carotenoid biosynthesis andlipid accumulation.This study provides new insights and a reference for the development of oil-producing strainswith high carotenoid yields through metabolic engineering.To our knowledge,similar studies have not beenreported in oil-producing red yeasts.

关 键 词：红冬孢酵母 RkCrtYB基因 过表达 类胡萝卜素 油脂 

分 类 号：Q786[生物学—分子生物学]