Production of isoorientin and isovitexin from luteolin and apigenin using coupled catalysis of glycosyltransferase and sucrose synthase
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Isoorientin and isovitexin, kinds of flavone C-glycosides, exhibit a number of biological properties. In this work, The C-glucosyltransferase (Gt6CGT) gene from Gentiana triflora was cloned and expressed in Escherichia coli BL21(DE3). The optimal activity of Gt6CGT was at pH 7.5 and 50° C. The enzyme was stable over pH range of 6.5–9.0, and had a 1-h half-life at 50° C. The Vmax for luteolin and apigenin was 21.1 nmol/min/mg and 31.7 nmol/min/mg, while the Km was 0.21 mM and 0.22 mM, respectively. Then, we developed an environmentally safe and efficient method for isoorientin and isovitexin production using the coupled catalysis of Gt6CGT and Glycine max sucrose synthase (GmSUS). By optimizing coupled reaction conditions, the titer of isoorientin and isovitexin reached 3820 mg/L with a corresponding molar conversion of 94.7% and 3772 mg/L with a corresponding molar conversion of 97.1%, respectively. The maximum number of UDP-glucose regeneration cycles (RCmax) reached 28.4 for isoorientin and 29.1 for isovitexin. The coupled catalysis reported herein represents a promising method to meet industrial requirements for large-scale isoorientin and isovitexin production in the future.
KeywordsIsoorientin Isovitexin Glycosyltransferase Sucrose synthase Coupled catalysis
This work was supported by the National Key R&D Program of China (2017YFD0600805), the National Natural Science Foundation of China (31570565), the Natural Science Foundation of the Jiangsu Province (BK20160929), the Open Foundation of Jiangsu Provincial Engineering Laboratory for Biomass Conversion and Process Integration (JPELBCPI2017002), and the Qing Lan Project and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The article does not contain any studies with human participants performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
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