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Updated: May 5, 2026

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

Published on: February 25, 2010

Biosíntesis Eficiente del Ginsenósido Raro Compuesto K Utilizando Pichia pastoris Modificada con β-Glucosidasa de

Yizi Luo^1,2, Pan Wang^1,2, Ruyao Wang^1,2

¹Engineering Research Center of Western Resource Innovation Medicine Green Manufacturing, Ministry of Education, School of Chemical Engineering, Northwest University, Xi'an 710069, China.

Journal of agricultural and food chemistry

|December 30, 2025

Abstract:

Sulfolobus solfataricus β-glucosidase (SS-bgly) is vital for converting rare ginsenoside compound K (CK), but suffers from stability and cost issues in practical applications. To tackle these, we developed a whole-cell immobilization method by genetically anchoring SS-bgly on Pichia pastoris GS115, using GPI-anchored GCW61 protein. The enhanced strain, Cbg61, underwent ARTP mutagenesis, yielding the improved Cbg61-T37 strain. This strain was further engineered to coexpress N-acetyltransferase MPR1 for oxidative stress resistance, resulting in Cbg61-T37-MPR1. Following fermentation optimization, the β-glucosidase activity of strain Cbg61-T37-MPR1 increased by 58.9% over that of strain Cbg61, with the process also resulting in a 93.79% conversion rate and a CK yield of 6.42 mg/mL. The strain exhibited high stability, retaining 63.17% efficiency after five cycles. This strategy of enzyme surface display, directed evolution, and reaction optimization significantly enhances CK production's practicality and economic feasibility.

Palabras clave:

ARTP MPR1 display en superficie celular ginsenósido raro CK β-glucosidasa

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