Astaxanthin overproduction in yeast by strain engineering and new gene target uncovering.
Astaxanthin overproduction in yeast by strain engineering and new gene target uncovering.
复制标题
通过菌株工程和新基因靶标的发现在酵母中过量生产虾青素
DOI：
10.1186/s13068-018-1227-4
复制
发表时间：
2018
影响因子：
6.3
通讯作者：
Yuan Y
中科院分区：
文献类型：
--
作者：
Jin J;Wang Y;Yao M;Gu X;Li B;Liu H;Ding M;Xiao W;Yuan Y
虾青素是一种天然类胡萝卜素，具有很强的抗氧化活性和商业价值。利用代谢工程微生物生产虾青素已成为一种很有前途的替代方法。尽管在异源模块和前体池的调整方面做了很大的努力，但这些非胡萝卜素产生菌的虾青素产量仍然不能令人满意地实现商业化，这表明除了在合理的代谢设计指导下有针对性地剪裁有限的靶标外，结合更多的全球干扰对虾青素生物合成系统的影响以及揭示新的分子机制似乎对进一步的开发更为关键。由于代谢工程和筛选诱变相结合是实现更多全球变异甚至发现更多分子靶标的有力工具，本研究将采用整合异源模块工程和常温等离子体诱变的综合方法来促进酿酒酵母虾青素的生产。从橙色农杆菌获得的新CrtZ菌株PC-1使虾青素产量提高到1.78倍，虾青素比率从66.6%提高到88.7%。通过ARTP诱变，虾青素的产量进一步提高了0.83倍(达到10.1 mg/g DCW)，这是迄今为止报道的酵母摇瓶水平上的最高产量。通过比较基因组学分析首次发现了与虾青素生物合成相关的三个潜在分子靶点(CSS1，YBR012W-BandDAN4)。值得注意的是，CSS1的个体缺失可以恢复ARTP诱变获得的虾青素产量的75.6%的提高，表明CSS1是一个非常有前途的分子靶标。在不添加诱导剂的情况下，最终在5-L发酵罐中获得了217.9 mg/L虾青素(虾青素率为89.4%，虾青素产量达到13.8g/gDCW)。结论通过对途径模块的合理设计和对宿主的随机诱变，本研究逐步提高了虾青素在酵母中的产量，达到了迄今报道的最高水平。这项工作不仅打破了在酵母中生产虾青素的上限，而且实现了关于类异戊二烯微生物过量生产的潜在分子靶标库。
BackgroundAstaxanthin is a natural carotenoid pigment with tremendous antioxidant activity and great commercial value. Microbial production of astaxanthin via metabolic engineering has become a promising alternative. Although great efforts have been conducted by tuning the heterologous modules and precursor pools, the astaxanthin yields in these non-carotenogenic microorganisms were still unsatisfactory for commercialization, indicating that in addition to targeted tailoring limited targets guided by rationally metabolic design, combining more globe disturbances in astaxanthin biosynthesis system and uncovering new molecular mechanisms seem to be much more crucial for further development. Since combined metabolic engineering with mutagenesis by screening is a powerful tool to achieve more global variations and even uncover more molecular targets, this study would apply a comprehensive approach integrating heterologous module engineering and mutagenesis by atmospheric and room temperature plasma (ARTP) to promote astaxanthin production inSaccharomyces cerevisiae.ResultsHere, compared to the strain with β-carotene hydroxylase (CrtZ) fromAlcaligenessp. strain PC-1, involving new CrtZ fromAgrobacterium aurantiacumenhanced astaxanthin yield to 1.78-fold and increased astaxanthin ratio to 88.7% (from 66.6%). Astaxanthin yield was further increased by 0.83-fold (to 10.1 mg/g DCW) via ARTP mutagenesis, which is the highest reported yield at shake-flask level in yeast so far. Three underlying molecular targets (CSS1,YBR012W-BandDAN4) associated with astaxanthin biosynthesis were first uncovered by comparative genomics analysis. To be noted, individual deletion ofCSS1can recover 75.6% improvement on astaxanthin yield achieved by ARTP mutagenesis, indicatingCSS1was a very promising molecular target for further development. Eventually, 217.9 mg/L astaxanthin (astaxanthin ratio was 89.4% and astaxanthin yield was up to 13.8 mg/g DCW) was obtained in 5-L fermenter without any addition of inducers.ConclusionsThrough integrating rational engineering of pathway modules and random mutagenesis of hosts efficiently, our report stepwise promoted astaxanthin yield to achieve the highest reported one in yeast so far. This work not only breaks the upper ceiling of astaxanthin production in yeast, but also fulfills the underlying molecular targets pools with regard to isoprenoid microbial overproductions.
登录
查看更多内容
影响因子：
6.4
作者：
Martín JF;Gudiña E;Barredo JL
通讯作者：
Barredo JL
影响因子：
6.4
作者：
Chen Y;Xiao W;Wang Y;Liu H;Li X;Yuan Y
通讯作者：
Yuan Y
DOI：
10.1007/978-1-4939-1363-3_1
发表时间：
2014-01-01
期刊：
YEAST GENETICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Gietz, R. Daniel
通讯作者：
Gietz, R. Daniel
开发裂殖壶菌生产二十二碳六烯酸的实时生物过程监测方法。
影响因子：
11.4
作者：
Guo, Dong-Sheng;Ji, Xiao-Jun;Huang, He
通讯作者：
Huang, He
影响因子：
3.9
作者：
Hua, Xiufu;Wang, Jun;Liu, Zheng
通讯作者：
Liu, Zheng