Evaluation and screening of efficient promoters to improve astaxanthin production in Xanthophyllomyces dendrorhous
Evaluation and screening of efficient promoters to improve astaxanthin production in Xanthophyllomyces dendrorhous
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DOI：
10.1007/s00253-014-5727-2
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发表时间：
2014-08-01
影响因子：
5
通讯作者：
Kondo, Akihiko
中科院分区：
文献类型：
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作者：
Hara, Kiyotaka Y.;Morita, Toshihiko;Kondo, Akihiko
虾青素是一种有价值的类胡萝卜素，广泛应用于水产养殖、食品、制药和化妆品行业。Xanthophyllomyces dendrorhous是一种合成类胡萝卜素的酵母菌株，其产生的虾青素作为其主要色素。虽然利用基因操作的代谢工程是提高虾青素产量的一种有价值的方法，但X。树状结构发育不良。在本研究中，三个已知的X.使用绿色荧光蛋白（GFP）作为表达水平指示蛋白，评价了树状结构、甘油-3-磷酸脱氢酶（gpd）启动子（Pgpd）、葡萄糖脱氢酶（gdh）启动子（Pgdh）和肌动蛋白（act）启动子（Pact）在过表达靶蛋白中的用途。与Pgpd和Pgdh相比，肌动蛋白启动子Pact显示出最高的GFP表达水平。另外，为了获得新的启动子，使目的蛋白在X.对13个候选启动子评估了树状的细胞内GFP强度。醇脱氢酶启动子Padh 4显示出比Pact更有效的GFP表达。crtE基因编码类胡萝卜素合成的限速酶的adh 4启动子下的过表达产生了约1.7倍，与对照菌株相比，细胞内的虾青素含量的增加。本研究中鉴定的启动子对提高X.树枝状的
Astaxanthin is a valuable carotenoid that is widely used in the aquaculture, food, pharmaceutical, and cosmetic industries. Xanthophyllomyces dendrorhous is a carotenoid-synthesizing yeast strain that produces astaxanthin as its main pigment. Although metabolic engineering using gene manipulation is a valuable way to improve astaxanthin production, a gene expression system for X. dendrorhous has been poorly developed. In this study, three known promoters of X. dendrorhous, glycerol-3-phosphate dehydrogenase (gpd) promoter (Pgpd), glucose dehydrogenase (gdh) promoter (Pgdh), and actin (act) promoter (Pact), were evaluated for use in the overexpression of target proteins using green fluorescence protein (GFP) as an expression level indicator protein. The actin promoter, Pact, showed the highest expression level of GFP when compared with Pgpd and Pgdh. Additionally, to obtain new promoters for higher expression of target protein in X. dendrorhous, intracellular GFP intensity was evaluated for 13 candidate promoters. An alcohol dehydrogenase promoter, Padh4, showed more efficient expression of GFP rather than Pact. Overexpression of crtE gene encoding rate-limiting enzyme of carotenoid synthesis under the adh4 promoter yielded an increase in intracellular astaxanthin content of about 1.7-fold compared with the control strain. The promoters identified in this study must be useful for improving carotenoids production in X. dendrorhous.