Genetic Modification of Mucor circinelloides for Canthaxanthin Production by Heterologous Expression of β-carotene Ketolase Gene.
Genetic Modification of Mucor circinelloides for Canthaxanthin Production by Heterologous Expression of β-carotene Ketolase Gene.
复制标题
DOI：
10.3389/fnut.2021.756218
复制
发表时间：
2021
影响因子：
5
通讯作者：
Song Y
中科院分区：
文献类型：
--
作者：
Naz T;Yang J;Nosheen S;Sun C;Nazir Y;Mohamed H;Fazili ABA;Ullah S;Li S;Yang W;Garre V;Song Y
角黄素是一种橙红色的叶黄素，具有较强的抗氧化活性和比胡萝卜素更高的生物利用度，主要用于食品、化妆品、水产养殖和制药工业。天然黄质市场的激增促使研究人员对异源宿主的基因工程进行黄质生产。卷枝毛霉（Mucor circinelloides）是一种产生β-胡萝卜素的二型真菌，是研究类胡萝卜素的模式生物。本研究以产黄嘌呤的M.通过将雨生红球藻β-胡萝卜素酮醇酶基因（bkt）在强启动子zrt 1的控制下整合到真菌基因组中，获得了卷枝藻菌株。首先，构建基本质粒以破坏crgA基因，crgA基因是胡萝卜素生物合成的负调控因子，导致大量的β-胡萝卜素产生，其通过酮醇酶的进一步酶促反应充当黄原胶的构建单元。该基因工程菌株产生显著量（576 ± 28 μg/g）的黄原胶，这是迄今为止在毛霉中报道的最高量。重组菌96 h后的菌体干重可达9.0 g/L以上。通过RT-qPCR分析过表达菌株中bkt的mRNA表达水平，与对照菌株相比，在24、48和72 h分别增加5.3、4.1和3倍。产黄嘌呤的M.本研究获得的一株类卷枝藻为进一步提高虾青素的生物技术生产奠定了基础，并为构建更有价值的类胡萝卜素，如虾青素，提供了一条有用的途径。
Canthaxanthin is a reddish-orange xanthophyll with strong antioxidant activity and higher bioavailability than carotenes, primarily used in food, cosmetics, aquaculture, and pharmaceutical industries. The spiking market for natural canthaxanthin promoted researchers toward genetic engineering of heterologous hosts for canthaxanthin production. Mucor circinelloides is a dimorphic fungus that produces β-carotene as the major carotenoid and is considered as a model organism for carotenogenic studies. In this study, canthaxanthin-producing M. circinelloides strain was developed by integrating the codon-optimized β-carotene ketolase gene (bkt) of the Haematococcus pluvialis into the genome of the fungus under the control of strong promoter zrt1. First, a basic plasmid was constructed to disrupt crgA gene, a negative regulator of carotene biosynthesis resulted in substantial β-carotene production, which served as the building block for canthaxanthin by further enzymatic reaction of the ketolase enzyme. The genetically engineered strain produced a significant amount (576 ± 28 μg/g) of canthaxanthin, which is the highest amount reported in Mucor to date. Moreover, the cell dry weight of the recombinant strain was also determined, producing up to more than 9.0 g/L, after 96 h. The mRNA expression level of bkt in the overexpressing strain was analyzed by RT-qPCR, which increased by 5.3-, 4.1-, and 3-folds at 24, 48, and 72 h, respectively, compared with the control strain. The canthaxanthin-producing M. circinelloides strain obtained in this study provided a basis for further improving the biotechnological production of canthaxanthin and suggested a useful approach for the construction of more valuable carotenoids, such as astaxanthin.
登录
查看更多内容
影响因子：
2.2
作者：
Gutierrez, Adrian;Lopez-Garcia, Sergio;Garre, Victorian
通讯作者：
Garre, Victorian
影响因子：
3.1
作者：
Navarro, E;Lorca-Pascual, JM;Ruiz-Vázquez, RM
通讯作者：
Ruiz-Vázquez, RM
卷枝毛霉基因改造DGLA生产细胞工厂的构建
影响因子：
6.4
作者：
Khan, Md Ahsanul Kabir;Yang, Junhuan;Song, Yuanda
通讯作者：
Song, Yuanda
影响因子：
--
作者：
Naz T;Nazir Y;Nosheen S;Ullah S;Halim H;Fazili ABA;Li S;Mustafa K;Mohamed H;Yang W;Song Y
通讯作者：
Song Y
影响因子：
5
作者：
Nicolas-Molina, Francisco E.;Navarro, Eusebio;Ruiz-Vazquez, Rosa M.
通讯作者：
Ruiz-Vazquez, Rosa M.