Identification of a novel glucuronyltransferase from Streptomyces chromofuscus ATCC 49982 for natural product glucuronidation
| Autor: | Jie Ren, Caleb Don Barton, Kathryn Eternity Sorenson, Jixun Zhan |
|---|---|
| Přispěvatelé: | Springer |
| Rok vydání: | 2022 |
| Předmět: |
Biological Products
food and beverages General Medicine Streptomyces chromofuscus Applied Microbiology and Biotechnology Streptomyces Flexible substrate specificity Kinetics Glucuronides Monoglucuronide Escherichia coli Diglucuronide Microbial glucuronyltransferase Glucuronosyltransferase Biomedical Engineering and Bioengineering Biotechnology |
| Zdroj: | Biological Engineering Faculty Publications |
| ISSN: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-022-11789-2 |
| Popis: | Glycosylation is an effective way to increase the polarity of natural products. UDP-glucuronyltransferases (UGTs) are commonly observed and extensively studied in phase II drug metabolism. However, UGTs in microorganisms are not well studied, which hampered the utilization of this type of enzyme in microbial glucuronidation of natural products. Screening of five actinomycete strains showed that Streptomyces chromofuscus ATCC 49982 can convert diverse plant polyphenols into more polar products, which were characterized as various glucuronides based on their spectral data. Analysis of the genome of this strain revealed a putative glucuronidation gene cluster that contains a UGT gene (gcaC) and two UDP-glucuronic acid biosynthetic genes (gcaB and gcaD). The gcaC gene was cloned and heterologously expressed in Escherichia coli BL21(DE3). Incubation of the purified enzyme with resveratrol and UDP-glucuronic acid led to the production of resveratrol-4′-O-β-D-glucuronide and resveratrol-3-O-β-D-glucuronide, allowing GcaC to be characterized as a flexible UGT. The optimal in vitro reaction pH and temperature for GcaC are 7.5 and 30 °C, respectively. Its activity can be stimulated by Ca2+, Mg2+, and Mn2+, whereas Zn2+, Cu2+, and Fe2+ showed inhibitory effects. Furthermore, GcaC has a broad substrate specificity, which can glucuronidate various substrates besides resveratrol, including quercetin, ferulic acid, vanillic acid, curcumin, vanillin, chrysin, zearalenone, and apigenin. The titers of resveratrol-4′-O-β-D-glucuronide and resveratrol-3-O-β-D-glucuronide in E. coli-GcaC were 78.381 ± 0.366 mg/L and 14.991 ± 0.248 mg/L from 114.125 mg/L resveratrol within 3 h. Therefore, this work provides an effective way to produce glucuronides of resveratrol and other health-benefitting natural products. |
| Databáze: | OpenAIRE |
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