食品与发酵工业2026，Vol.52Issue(11)：1-9,9.DOI:10.13995/j.cnki.11-1802/ts.044229
毕赤酵母表达α-L-鼠李糖苷酶的发酵工艺优化及酶固定化研究
Optimization of fermentation process and immobilization of α-L-rhamnosidase expressed by Pichia pastoris
摘要
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 conver-sion 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,in-dicated excellent storage stability.These findings provide both technical and theoretical foundations for the industrial production of α-L-rh-amnosidase and efficient biotransformation of high-value flavonoid compounds.关键词
α-L-鼠李糖苷酶/毕赤酵母/发酵工艺优化/酶固定化/芦丁转化Key words
α-L-rhamnosidase/Pichia pastoris/fermentation process optimization/enzyme immobilization/rutin biotransformation引用本文复制引用
芦星淼,朱清泉,侯飞鸿,杨超豪,任晓菲,路福平,刘夫锋..毕赤酵母表达α-L-鼠李糖苷酶的发酵工艺优化及酶固定化研究[J].食品与发酵工业,2026,52(11):1-9,9.基金项目
国家重点研发计划项目(2024YFA0917900) （2024YFA0917900）
国家自然科学基金项目(32272269) （32272269）
天津市合成生物技术创新能力提升行动项目(TSBICIP-PTJJ-010) （TSBICIP-PTJJ-010）
