Food and Fermentation Industries >

2026 , Vol. 52 >Issue 11: 1 - 9

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.044229

Optimization of fermentation process and immobilization of α-L-rhamnosidase expressed by Pichia pastoris

  • LU Xingmiao ,
  • ZHU Qingquan ,
  • HOU Feihong ,
  • YANG Chaohao ,
  • REN Xiaofei ,
  • LU Fuping ,
  • LIU Fufeng
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  • (College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, China)

Received date: 2025-08-14

  Revised date: 2025-10-29

  Online published: 2026-06-24

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Abstract

α-L-Rhamnosidase is a glycosidase capable of specifically hydrolyzing terminal rhamnose residues in flavonoids, widely used in food and pharmaceutical industries.However, its industrial application faces challenges of high cost and poor stability.Thus, this study optimized fermentation conditions and employed immobilization techniques to enhance enzyme activity and stability using a previously constructed recombinant Pichia pastoris strain GS115/pPIC9K-Rha-gcw61.The results showed that using BMGY as the medium, with 1.59% glycerol, 2.49% (NH4)2SO4 (mass fraction), and 2.90% inoculum (volume fraction), yield the highest enzyme activity of 170 U/mL in the fermentation broth.Bioreactor cultivation further increased enzyme activity to 1 380 U/mL after 86 h at 30% dissolved oxygen, which is 8.12-fold compared to shake-flask cultures.The enzyme solution was used to convert rutin into isoquercitrin, the conversion rate exceeded 90% within 24 h for rutin mass concentrations below 75 g/L, while higher concentrations (100, 125, and 150 g/L) required 48 h to achieve similar conversion rates.The optimal temperature and pH for the immobilized α-L-rhamnosidase were 60 ℃ and 5.0, respectively.Furthermore, the immobilized enzyme retained more than 70% of its initial activity after storage at 4 ℃ for 50 days, indicated excellent storage stability.These findings provide both technical and theoretical foundations for the industrial production of α-L-rhamnosidase and efficient biotransformation of high-value flavonoid compounds.

Key words： α-L-rhamnosidase; Pichia pastoris; fermentation process optimization; enzyme immobilization; rutin biotransformation

Cite this article

LU Xingmiao , ZHU Qingquan , HOU Feihong , YANG Chaohao , REN Xiaofei , LU Fuping , LIU Fufeng . Optimization of fermentation process and immobilization of α-L-rhamnosidase expressed by Pichia pastoris[J]. Food and Fermentation Industries, 2026 , 52(11) : 1 -9 . DOI: 10.13995/j.cnki.11-1802/ts.044229

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