Combinatorial enzymatic catalysis for bioproduction of ginsenoside Compound K

Abstract

Enzymatic catalysis provides a clean, efficient, and stable solution for the industrial preparation of Ginsenoside Compound K (CK). However, exploring high-efficiency enzymes and establishing catalytic processes remain challenging. Here, we report Aspergillus tubingensis JE0609 with the ability to efficiently transform protopanaxadiol-type ginsenosides into CK. Our transcriptome analysis and peptide mass fingerprinting reveal that three ginsenoside hydrolases BG07, BG19, and BG23 are present in JE0609. Notably, BG23 is highly expressed in Pichia pastoris with a yield of 235.73 U/mL (pNPG), with a broad adaptation to acidic (pH 4.5 to 7.0) and thermophilic (temperature < 50 °C) conditions. Inspired by the above outcomes, we establish a one-pot combinatorial enzyme-catalyzed strategy for conversion of Rb1, Rb2, Rb3, and Rc into CK, with a yield of 396.7 mg·L-1·h-1, which is the highest one by now. We argue that this combinatorial enzymatic catalysis strategy opens the probability of bioproduction of CK and other ginsenosides.