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Improved Astaxanthin Production with Corynebacterium glutamicum by Application of a Membrane Fusion Protein

机译:应用膜融合蛋白改善谷氨酸棒杆菌生产虾青素的能力

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摘要

Astaxanthin is one of the strongest natural antioxidants and a red pigment occurring in nature. This C40 carotenoid is used in a broad range of applications such as a colorant in the feed industry, an antioxidant in cosmetics or as a supplement in human nutrition. Natural astaxanthin is on the rise and, hence, alternative production systems are needed. The natural carotenoid producer is a potent host for industrial fermentations, such as million-ton scale amino acid production. In , astaxanthin production was established through heterologous overproduction of the cytosolic lycopene cyclase CrtY and the membrane-bound β-carotene hydroxylase and ketolase, CrtZ and CrtW, in previous studies. In this work, further metabolic engineering strategies revealed that the potential of this GRAS organism for astaxanthin production is not fully exploited yet. It was shown that the construction of a fusion protein comprising the membrane-bound β-carotene hydroxylase and ketolase (CrtZ~W) significantly increased astaxanthin production under high glucose concentration. An evaluation of used carbon sources indicated that a combination of glucose and acetate facilitated astaxanthin production. Moreover, additional overproduction of cytosolic carotenogenic enzymes increased the production of this high value compound. Taken together, a seven-fold improvement of astaxanthin production was achieved with 3.1 mg/g CDW of astaxanthin.
机译:虾青素是最强的天然抗氧化剂之一,也是自然界中的红色素。这种C40类胡萝卜素的用途广泛,例如饲料工业中的着色剂,化妆品中的抗氧化剂或人体营养补充剂。天然虾青素正在增加,因此需要替代生产系统。天然的类胡萝卜素生产者是工业发酵的强大宿主,例如百万吨级氨基酸的生产。在以前的研究中,虾青素的产生是通过胞质番茄红素环化酶CrtY和膜结合的β-胡萝卜素羟化酶和酮醇酶,CrtZ和CrtW的异源过量产生而建立的。在这项工作中,进一步的代谢工程学策略表明该GRAS生物体生产虾青素的潜力尚未得到充分利用。结果表明,在高葡萄糖浓度下,构建包含膜结合的β-胡萝卜素羟化酶和酮醇酶(CrtZ〜W)的融合蛋白可以显着提高虾青素的产量。对用过的碳源的评估表明,葡萄糖和乙酸盐的组合促进了虾青素的产生。此外,胞质类胡萝卜素生成酶的额外过量生产增加了这种高价值化合物的生产。两者合计,用3.1mg / g CDW的虾青素可以将虾青素的产量提高七倍。

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