巴斯德毕赤酵母(Pichia pastoris)是工业上生产异源蛋白的良好平台,且具有合成β-胡萝卜素的潜力,为推进β-胡萝卜素工业化生产,克隆来自锁掷酵母(Sporidiobolus pararoseus)的类胡萝卜素相关基因,并整合到巴斯德毕赤酵母基因组中,产生类胡萝卜素生产菌株Pp-EYBI。通过优化β-胡萝卜素合成限速酶基因拷贝数并过表达MVA限速酶基因,增强前体物质的供应及流向β-胡萝卜素合成的通量,最终获得菌株Pp-EYBI+(YB)3H,其β-胡萝卜素产量为3.7 mg/gDCW。该研究首次克隆来自锁掷酵母的类胡萝卜素相关基因并实现其在毕赤酵母中的表达,为异源合成类胡萝卜素提供了新的思路,且有望通过巴斯德毕赤酵母实现工业生产β-胡萝卜素。
In order to promote the industrial production of β-carotene, carotenoid-related genes were cloned from Sporidiobolus pararoseus and integrated into the genome of Pichia pastoris and a carotenoid-producing strain Pp-EYBI was constructed. Meanwhile, the gene copy number of the rate-limiting enzyme for β-carotene synthesis was optimized and the gene of MVA rate-limiting enzyme was over-expressed to enhance the supply of precursor materials and the flux to β-carotene synthesis. The strain Pp-EYBI+(YB)3H was finally obtained, which yielded as high as 3.7 mg/g (dry cell weight) β-carotene. This lays a foundation for the industrial production of β-carotene by P. pastoris.
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