Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 68, Issue 35, Pages 9513-9523Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c03689
Keywords
UDP-rhamnose; cellobiose phosphorolysis pathway; UDP-rhamnose synthase; rhamnosylation; NADPH regenerator; quercitrin
Funding
- National Key R&D Program of China [2017YFD0600805]
- National Natural Science Foundation of China [21978135]
- Open Foundation of Jiangsu Provincial Engineering Laboratory for Biomass Conversion and Process Integration [JPELBCPI2017002]
- Qing Lan Project
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
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UDP-rhamnose is the main type of sugar donor and endows flavonoids with special activity, selectivity, and pharmacological properties by glycosylation. In this study, several UDP-glucose synthesis pathways and UDP-rhamnose synthases were screened to develop an efficient UDP-rhamnose biosynthesis pathway in Escherichia coli. Maximal UDP-rhamnose production reached 82.2 mg/L in the recombinant strain by introducing the cellobiose phosphorolysis pathway and Arabidopsis thaliana UDP-rhamnose synthase (AtRHM). Quercitrin production of 3522 mg/L was achieved in the recombinant strain by coupling the UDP-rharnnose generation system with A. thaliana rhamnosyltransferase (AtUGT78D1) to recycle UDP-rharnnose. To further increase UDP-rhamnose supply, an NADPH-independent fusion enzyme was constructed, the UTP supply was improved, and NADPH regenerators were overexpressed in vivo. Finally, by optimizing the bioconversion conditions, the highest quercitrin production reached 7627 mg/L with the average productivity of 141 mg/(L h), which is the highest yield of quercitrin and efficiency of UDPrhamnose supply reported to date in E. coli. Therefore, the method described herein for the regeneration of UDP-rhamnose from cellobiose may be widely used for the rhamnosylation of flavonoids and other bioactive substances.
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