期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 71, 期 41, 页码 15204-15212出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.3c04419
关键词
chlorogenicacid; Escherichia coli; shikimate; cofactor engineering; YdiB; HpaBC
A de novo biosynthesis pathway for chlorogenic acid was constructed in Escherichia coli via modular engineering, which increased metabolic flux and cofactor supply for efficient production of chlorogenic acid.
Chlorogenic acid is a natural phenolic compound widely used in the food and daily chemical industries. Compared to plant extraction, microbial cell factories provide a green and sustainable production method for the production of chlorogenic acid. However, complex metabolic flux distribution and potential byproducts limited the biosynthesis of chlorogenic acid in microorganisms. A de novo biosynthesis pathway for chlorogenic acid was constructed in Escherichia coli via modular engineering. Increasing the shikimate pathway flux greatly promoted chlorogenic acid production, and the influence of pyruvate metabolism on chlorogenic acid synthesis was also explored. The supply of cofactors for the key enzymes quinate/shikimate 5-dehydrogenase (YdiB) and 4-hydroxyphenylacetate 3-monooxygenase (HpaBC) was enhanced by a cofactor regeneration system. Furthermore, mutants of YdiB were verified for chlorogenic acid production in vivo. Chlorogenic acid browning occurred when the buffer pH of the buffer exceeded 6.0, but two-stage pH control achieved a chlorogenic acid titer of 2789.2 mg/L in a 5 L fermenter, the highest reported to date. This study provided a strategy for the efficient production of chlorogenic acid from simple carbon sources.
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