提高法菲黑马霉葡萄糖氧化酶异源表达的联合策略

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2025-07-05 DOI:10.1016/j.bej.2025.109856

Qingzhu Wang , Lulu Chen , Jia Zheng , Bowen Ke , Yi’an Chen , Yaping Zhang , Shuangyan Han

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引用次数: 0

摘要

葡萄糖氧化酶（EC 1.1.3.4， GOD）是一种重要的工业酶，在食品、制药、临床和化学工业中有着广泛的应用。本研究从cristatus曲霉中鉴定出一种葡萄糖氧化酶cGOD，并在Komagataella phaffii中异源表达。生化表征表明，cGOD在40 °C和pH 6.0条件下具有最佳活性。通过优化启动子和信号肽、扩增基因拷贝数和调节分泌途径等组合策略来提高其产量。结果表明，摇瓶培养的细胞外cGOD活性增加到967.23 U/mL。值得注意的是，在15 L补料间歇发酵罐中，cGOD酶活性达到11,655 U/mL，超过了先前报道的GOD活性水平。该研究的成功鉴定和cGOD基因的高效表达，为开发高效产酶微生物菌株提供了有价值的见解。

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Combined strategies to improve the heterologous expression of glucose oxidase from Aspergillus cristatus in Komagataella phaffii

Glucose oxidase (EC 1.1.3.4, GOD) is a key industrial enzyme with broad applications across the food, pharmaceutical, clinical, and chemical industries. In this study, a glucose oxidase, designated cGOD, was identified from Aspergillus cristatus and heterologously expressed in Komagataella phaffii. Biochemical characterization revealed that cGOD exhibited optimal activity at 40 °C and pH 6.0. To enhance its production, a combinatorial strategy was implemented, involving optimization of promoter and signal peptide, amplification of gene copy number, and modulation of the secretory pathway. As a result, extracellular cGOD activity increased to 967.23 U/mL in shake flask culture. Notably, in a 15 L fed-batch fermenter, the cGOD enzyme activity reached 11,655 U/mL, surpassing previously reported GOD activity levels. The successful identification and efficient expression of cGOD from A. cristatus provide valuable insights for the development of high-yield microbial strains for industrial enzyme production.

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来源期刊

Biochemical Engineering Journal 工程技术-工程：化工

CiteScore

7.10

自引率

5.10%

发文量

380

审稿时长

34 days

期刊介绍： The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology. The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields: Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics Biosensors and Biodevices including biofabrication and novel fuel cell development Bioseparations including scale-up and protein refolding/renaturation Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells Bioreactor Systems including characterization, optimization and scale-up Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis Protein Engineering including enzyme engineering and directed evolution.