Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, PR China.
Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, PR China; Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University, Hangzhou 310027, PR China.
Metab Eng. 2015 Mar;28:8-18. doi: 10.1016/j.ymben.2014.11.007. Epub 2014 Dec 2.
Balanced utilization of metabolic intermediates and controllable expression of genes in biosynthetic pathways are key issues for the effective production of value-added chemicals in microbes. An inducer/repressor-free sequential control strategy regulated by glucose concentration in the growth environment was proposed to address these issues, and its efficiency was validated using heterologous beta-carotenoid biosynthesis in Saccharomyces cerevisiae as an example. Through sequential control of the downstream, upstream, and competitive pathways of farnesyl diphosphate (FPP), the crucial metabolic node in the biosynthesis of terpenoids, in a predetermined order, a carotenoid production of 1156 mg/L (20.79 mg/g DCW) was achieved by high-cell density fermentation. Quantitative PCR analysis of the regulated genes demonstrated that the transcription patterns were controlled in a sequential manner as expected. The inducer/repressor-free nature of this strategy offers a both practical and economically efficient approach to improved biosynthetic production of value-added chemicals.
在微生物中有效生产高附加值化学品的关键问题是代谢中间产物的平衡利用和生物合成途径中基因的可控表达。本研究提出了一种受生长环境中葡萄糖浓度调控的无诱导剂/抑制剂的顺序控制策略，并以异源β-胡萝卜素生物合成为例验证了其效率。通过顺序控制法控制法尼焦磷酸（FPP）的下游、上游和竞争途径，这是萜类化合物生物合成中的关键代谢节点，在预定的顺序下，通过高密度发酵实现了 1156mg/L（20.79mg/gDCW）的类胡萝卜素产量。对调控基因的定量 PCR 分析表明，转录模式按预期的顺序进行控制。该策略的无诱导剂/抑制剂特性为提高高附加值化学品的生物合成生产提供了一种实用且经济高效的方法。
