Efficient Production of 2,5-Diketo-D-gluconic Acid by Reducing Browning Levels During Gluconobacter oxydans ATCC 9937 Fermentation.
Efficient Production of 2,5-Diketo-D-gluconic Acid by Reducing Browning Levels During Gluconobacter oxydans ATCC 9937 Fermentation.
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DOI：
10.3389/fbioe.2022.918277
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发表时间：
2022
影响因子：
5.7
通讯作者：
Zhou, Jingwen
中科院分区：
文献类型：
--
作者：
Li, Guang;Shan, Xiaoyu;Zeng, Weizhu;Yu, Shiqin;Zhang, Guoqiang;Chen, Jian;Zhou, Jingwen
d -葡萄糖通过2,5-二酮-d -葡萄糖酸（2,5- dkg）途径直接生成2-酮- l -谷醛酸（2- klg，维生素C的前体）。2,5- dkg是该反应的主要限速因子，目前相关研究较少。本研究建立了一种更准确的2,5- dkg定量方法，并用于筛选氧化革菌ATCC9937作为生产2,5- dkg的基质菌株。结合代谢物谱分析和生产菌株的敲除和过表达，确定了2,5- dkg的非酶褐变是导致目标化合物低产量的主要因素。通过优化发酵工艺，将发酵时间缩短至48 h， 2,5- dkg产量达到50.9 g/L，比对照组提高139.02%。有效地消除褐变，减少2,5- dkg的降解，有助于提高2,5- dkg向2- klg的转化，最终建立一步d -葡萄糖到2- klg的发酵途径。
D-Glucose directly generates 2-keto-L-gulonic acid (2-KLG, precursor of vitamin C) through the 2,5-diketo-D-gluconic acid (2,5-DKG) pathway. 2,5-DKG is the main rate-limiting factor of the reaction, and there are few relevant studies on it. In this study, a more accurate quantitative method of 2,5-DKG was developed and used to screen G. oxydans ATCC9937 as the chassis strain for the production of 2,5-DKG. Combining the metabolite profile analysis and knockout and overexpression of production strain, the non-enzymatic browning of 2,5-DKG was identified as the main factor leading to low yield of the target compound. By optimizing the fermentation process, the fermentation time was reduced to 48 h, and 2,5-DKG production peaked at 50.9 g/L, which was 139.02% higher than in the control group. Effectively eliminating browning and reducing the degradation of 2,5-DKG will help increase the conversion of 2,5-DKG to 2-KLG, and finally, establish a one-step D-glucose to 2-KLG fermentation pathway.
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