光照刺激卷枝毛霉β-胡萝卜素积累的转录组学分析  

Transcriptome analysis ofβ-carotene accumulation by light exposure in Mucor circinelloides

作　　者：党文瑞 李鹤群 袁洪娟 周凤艳 张怀渊 DANG Wenrui;LI Hequn;YUAN Hongjuan;ZHOU Fengyan;ZHANG Huaiyuan(College of Agricultural Engineering and Food Science,Shandong University of Technology,Zibo 255000,China;All Mine Testing Co.,Ltd.,Heze 274008,China)

机构地区：[1]山东理工大学农业工程与食品科学学院,淄博255000 [2]奥迈检测有限公司,菏泽274008

出　　处：《农业工程学报》2024年第19期300-309,共10页Transactions of the Chinese Society of Agricultural Engineering

基　　金：山东省自然科学基金面上项目(ZR2022MC108)。

摘　　要：为明确光照刺激微生物合成β-胡萝卜素具体代谢过程中的分子机制,该研究使用超高效液相色谱法及Illumina Hiseq 2000平台分别对卷枝毛霉WJ11光照刺激后β-胡萝卜素积累及代谢通路中的基因变化及功能注释进行转录组学分析。结果表明光照后细胞干质量比黑暗条件稍高,葡萄糖的消耗速率更快且脂质含量明显提高,达到细胞干质量的35.3%,而光照后β-胡萝卜素合成急剧增加,持续光照1 440 min时β-胡萝卜素质量分数达到最大值458.3μg/g,是黑暗条件的2.1倍。分别收取光照与黑暗条件下不同时间点的样品进行转录组学分析,结果表明光照刺激后120 min糖酵解途径增强,积累的丙酮酸在细胞质中转化为乙醛并激活细胞质中丙酮酸-乙醛-乙酸循环,从而提供更多的前体乙酰辅酶A,甲羟戊酸途径基因表达量增强,使得β-胡萝卜素合成剧烈增加。而光照1 440 min后脂质和β-胡萝卜素合成趋于稳定,推测乙酰辅酶A有所积累导致乙酰辅酶A乙酰转移酶活性增强甲羟戊酸途径活性反而下降,β-胡萝卜素积累有所下降。此外乙酰辅酶A合成酶和ATP:柠檬酸裂解酶的表达量下降,使得碳源更多流向三羧酸循环。3个与光响应相关的光受体蛋白-1转录水平在光照120 min后均发生显著下调,推测其是光照刺激β-胡萝卜素合成的关键调控蛋白。研究结果为β-胡萝卜素合成分子机制研究及深化β-胡萝卜素遗传改造提供了理论支持。β-carotene is one of the most diverse and effective antioxidants in natural fat-soluble pigments in the food,pharmaceutical,and cosmetic industries.Much attention has been attracted in recent years,due to its potential antibacterial properties and anticancer agents.Among them,microbial synthesis can be expected to serve as the ever-increasingly important source ofβ-carotene,due to the lower cost,higher safety,and easy large-scale production,compared with animal and plant extraction and industrial synthesis.Light stimulation of carotenoid synthesis is also the earliest and most studied regulatory mechanism in fungi.The genome of Mucor circinelloides can be better used in theβ-carotene synthesis mechanism,due to the efficient genetic transformation,gene replacement,and silencing,as well as the ability to express foreign genes.This study aims to clarify the molecular mechanism in the specific metabolic process ofβ-carotene synthesis,where the microorganisms were stimulated by light.A systematic transcriptomic analysis was then made using high-performance liquid chromatography and Illumina Hiseq 2000.The gene changes and functional annotation was determined inβ-carotene accumulation and metabolic pathway of Mucor circinelloides WJ11 after light stimulation,respectively.The fermentation results showed that the cell dry weight was slightly higher under light exposure than under dark conditions,the maximum of which was up to 14.8 g/L under light exposure for 1440 min.There was a high rate of glucose consumption.The concentration of residual glucose was about 7.0 g/L in the medium at the late fermentation stage.In addition,the lipid content significantly increased,reaching 35.3%of the dry weight of cells.β-carotene was at the lower level under the two cultures before light.There was no difference in production.Mucor circinelloides WJ11 accumulated the maximum pigment amount under 1440 min of continuous light.The pigment amounts under light and dark conditions were 458.3 and 218.7μg/g,respectively,and then theβ-ca

关 键 词：卷枝毛霉 光照刺激 Β-胡萝卜素 转录组学 光受体蛋白-1 

分 类 号：TS201.3[轻工技术与工程—食品科学]