Complete Biotransformation of Protopanaxadiol-Type Ginsenosides into 20-O-β-Glucopyranosyl-20(S)-protopanaxadiol by Permeabilized Recombinant Escherichia coli Cells Coexpressing β-Glucosidase and Chaperone Genes

Abstract

The ginsenoside 20-O-β-glucopyranosyl-20(S)-protopanaxadiol or compound K is an essential ingredient in functional food, cosmetics, and traditional medicines. However, no study has reported the complete conversion of all protopanaxadiol (PPD)-type ginsenosides from ginseng extract into compound K using whole-cell conversion. To increase the production of compound K from ginseng extract using whole recombinant cells, the β-glucosidase enzyme from Caldicellulosiruptor bescii was coexpressed with a chaperone expression system (pGro7), and the cells expressing the coexpression system were permeabilized with ethylenediaminetetraacetic acid. The permeabilized cells carrying the chaperone coexpression system showed a 2.6-fold increase in productivity and yield as compared with nontreated cells, and completely converted all PPD-type ginsenosides from ginseng root extract into compound K with the highest productivity among the results reported so far. Our results will contribute to the industrial biological production of compound K.

Complete Bioconversion of Protopanaxadiol-Type Ginsenosides to Compound K by Extracellular Enzymes from the Isolated Strain Aspergillus tubingensis

“…The dried residue was treated by adding methanol. The concentrations of ginsenosides were determined using an Agilent 1100 Infinity high-performance liquid chromatography (HPLC) system (Santa Clara, CA, USA) equipped with an ultraviolet detector at a wavelength of 203 nm and an octadecyl-silica column, which was eluted at a flow rate of 1 mL/min with the solution by mixing a concentration of acetonitrile dissolved in H 2 O (v/v) with linear change from 30 to 60% for 20 min, 60 to 90% for 10 min, 90 to 30% for 5 min, and with a constant 30% for 10 min 13 The column temperature was maintained at 40 °C.…”

“…Compound K is only produced from the hydrolysis of sugar moieties in the major ginsenosides in the ginseng extract by biological methods, including fermentation, whole cell conversion, and enzymatic conversion. − Among these transformations, enzymatic conversion exhibits the highest production of compound K. The enzymatic conversion has been performed using recombinant enzymes, − commercial recombinant enzymes, , and native enzymes. − The conversion using recombinant enzymes showed the highest production of compound K than other enzymatic conversions. − However, the application of recombinant enzymes to food is strictly limited …”

Improved production of deglucosylated platycodin D from saponins from balloon flower leaf by a food-grade enzyme using high hydrostatic pressure

“…(Shin et al, 2019) The biological activity of saponins is improved by their deglycosylation, since they are more readily absorbed in the human gastrointestinal tract due to the lower molecular weight and better hydrophobicity (Park et al, 2010;Shin and Oh, 2016). Therefore, various methods such as heating (Kim et al, 2000), acid treatment (Bae et al, 2004), and biotransformation (Kim et al, 2019a;Shin et al, 2018) have been used to deglycosylate saponins. Among these, the enzymatic biotransformation of platycosides shows the highest selectivity and productivity, which has been demonstrated in diverse studies using a crude enzyme from Aspergillus niger (Wie et al, 2007), β-galactosidase from Aspergillus oryzae (Ha et al, 2010), β-glucosidase from Aspergillus usamii (Ahn et al, 2018), recombinant β-glucosidases from Caldicellulosiruptor bescii (Kil et al, 2019), Caldicellulosiruptor owensensis (Shin et al, 2019), and Dictyoglomus turgidum (Kang et al, 2019), and commercial enzymes, such as cellulase (Ha et al, 2010), laminarinase (Jeong et al, 2014), snailase (Li et al, 2012), pectinase (Ju et al, 2020), and cytolase (Shin et al, 2020).…”

Biocatalytic Synthesis of a Novel Bioactive Ginsenoside Using UDP-Glycosyltransferase from Bacillus subtilis 168

“…Ginseng (Panax ginseng) is the most famous medicinal-edible plant recorded in the Chinese Pharmacopoeia [1,2]. This precious perennial herb is crowned the "King of All Herbs" [3].…”