法夫酵母中产类胡萝卜素特性与其合成相关基因表达关系的研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.016

生物技术通报 ›› 2016, Vol. 32 ›› Issue (2): 123-130.doi: 10.13560/j.cnki.biotech.bull.1985.2016.02.016

法夫酵母中产类胡萝卜素特性与其合成相关基因表达关系的研究

郑晨华¹, 杜希萍¹, 李利君¹, 李天丽¹, 曹樱¹, 倪辉^1,2

1.集美大学食品与生物工程学院，厦门 361021；2.福建省食品微生物与酶工程重点实验室，厦门 361021

收稿日期:2015-04-30 出版日期:2016-02-24 发布日期:2016-02-25
作者简介:郑晨华，男，硕士，研究方向：微生物遗传育种；E-mail：zch52840@163.com
基金资助:
国家自然科学基金项目(20702019)，福建省自然科学基金项目(2012J01137)，集美大学校科研启动金项目(ZQ2007009)

The Relationship Between Characteristics of Yielding Carotenoid and Expression of Carotenogenic Genes in Phaffia rhodozyma

ZHENG Chen-hua¹, DU Xi-ping¹, LI Li-jun¹, LI Tian-li¹, CAO Ying¹, NI Hui^1,2

(1.College of Food and Biological Engineering，Jimei University，Xiamen 361021；2.Key Laboratory of Food Microbiology and Enzyme Engineering of Fujian Province，Xiamen 361021)

Received:2015-04-30 Published:2016-02-24 Online:2016-02-25

摘要/Abstract

摘要： 为研究法夫酵母中类胡萝卜素合成代谢调控机理，通过高效液相色谱法和实时定量PCR技术分别对JMU-VDL668和JMU-N3法夫酵母菌株中类胡萝卜素和类胡萝卜素合成相关基因的表达差异进行分析，并研究了在JMU-VDL668菌株培养过程中添加β-胡萝卜素对其产虾青素的影响。结果显示，法夫酵母类胡萝卜素代谢途径的关键酶crtE, crtYB, crtI和crtS基因在JMU-VDL668菌株中的表达水平整体低于JMU-N3菌株，而JMU-N3菌株中没有检测到crtR基因的表达。在JMU-VDL668菌株培养过程中添加β-胡萝卜素发现，在72 h和96 h添加时能促进虾青素的合成，其中96 h添加时所得虾青素比对照组提高了83%。结果表明，这两株菌的产类胡萝卜素特性与类胡萝卜素合成相关基因的表达量存在一定的正相关关系，且JMU-VDL668菌株中总类胡萝卜素细胞产率在一定范围内，可能存在反馈调节机制。

关键词: 法夫酵母, 虾青素, 实时定量PCR, 代谢调控

Abstract: In order to study the metabolic regulation mechanism of carotenoid synthesis in Phaffia rhodozyma, the carotenoid biosynthesis characteristics and the expression differences of carotenogenic genes in different strains of P.rhodozyma, JMU-VDL668 and JMU-N3 were analyzed by HPLC and real-time qPCR, respectively.The effects of adding β-carotene during the culture of strain JMU-VDL668 to the yield of astaxanthin were also analyzed.The results showed that, the expression levels of the carotenogenic genes crtE, crtI, crtYB and crtS of key enzymes in the metabolic pathway of carotenoid in P.rhodozyma were lower in JMU-VDL668 than JMU-N3, even the expression of crtR was not detected in JMU-N3.The synthesis of astaxanthin increased when β-carotene was added at 72 h and 96 h of culturing JMU-VDL668, and the astaxanthin increased 83% while adding β-carotene at 96 h compared to the control group.The results indicated that the characteristics of yielding carotenoid was positively correlated to the expression level of carotenogenic genes.Moreover, the productivity of cells for total carotenoid in JMU-VDL668 was in a certain range, suggesting that there was a feedback regulation.

Key words: Phaffia rhodozyma, astaxanthin, real-time qPCR, metabolic regulation

郑晨华, 杜希萍, 李利君, 李天丽, 曹樱, 倪辉. 法夫酵母中产类胡萝卜素特性与其合成相关基因表达关系的研究[J]. 生物技术通报, 2016, 32(2): 123-130.

ZHENG Chen-hua, DU Xi-ping, LI Li-jun, LI Tian-li, CAO Ying, NI Hui. The Relationship Between Characteristics of Yielding Carotenoid and Expression of Carotenogenic Genes in Phaffia rhodozyma[J]. Biotechnology Bulletin, 2016, 32(2): 123-130.

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法夫酵母中产类胡萝卜素特性与其合成相关基因表达关系的研究

The Relationship Between Characteristics of Yielding Carotenoid and Expression of Carotenogenic Genes in Phaffia rhodozyma

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