Evolutionary engineering of Phaffia rhodozyma for astaxanthin-overproducing strain

Evolutionary engineering is a novel whole- genome wide engineering strategy inspired by natural evolution for strain improvement. Astaxanthin has been widely used in cosmetics, pharmaceutical and health care food due to its capability of quenching active oxygen. Strain improvement ofPhaffia rhodozyma, one of the main sources for natural astaxanthin, is of commercial interest for astaxanthin production. In this study a selection procedure was developed for adaptive evolution of P. rhodozyma strains under endogenetic selective pressure induced by additive in environmental niches. Six agents, which can induce active oxygen in cells, were added to the culture medium respectively to produce selective pressure in process of evolution. The initial strain, P. rhodozyma AS2-1557, was mutagenized to acquire the initial strain population, which was then cultivated for 550 h at selective pressure and the culture was transferred every 48h. Finally, six evolved strains were selected after 150 generations of evolution. The evolved strains produced up to 48.2% more astaxanthin than the initial strain. Our procedure may provide a promising alternative for improvement of highproduction strain.

微生物进化工程育种技术进展与展望

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超突变细胞在进化工程中的开发与应用

通过进化工程技术改造微生物细胞的生理表型是生物技术和生物炼制领域的重要研究方向,但是现阶段的各种进化工程技术面临效率低或连续性差的问题。超突变细胞能够进行自发、连续的胞内诱变,将其应用于进化工程技术能够实现连续、高效的菌种改造。本文详细介绍了自然界中超突变细胞产生的遗传机制和相应的人工超突变细胞的构建策略,以及应用此类超突变系统在微生物细胞生理性能改造和蛋白质突变体文库高效构建上取得的进展。随着相关领域认识和技术的加强,未来人工超突变细胞的构建将继续向着不同维度上可控性、靶向性不断提高的方向发展,从而为细胞和蛋白的改造提供更强更优的进化动力。