Recombinant beta-galactosidase derived from Enterobacter cloacae Zjut HJ2001 for efficient biotransformation of galactooligosaccharides

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2024-09-25 DOI:10.1016/j.bej.2024.109514

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引用次数: 0

Abstract

Galactooligosaccharides (GOS), as a type of prebiotics, have excellent physical and chemical properties, and can be widely used in the pharmaceutical and food fields. Recently, the microbial β-galactosidases have gained widespread attentions in industrial GOS production. However, most β-galactosidases from microorganisms have low transgalactosylation activity, resulting in poor GOS yield of enzymatic transformation from lactose. In this paper, a brand new β-galactosidase derived from Enterobacter cloacae Zjut HJ2001 was screened out from soil, and successfully overexpressed, characterized, and mutated by combinatorial alanine-scanning and site-saturation mutagenesis. Compared to the yield of 51.73 % obtained by wild-type β-galactosidase with lactose concentration of 380 g/L, the obtained mutant β-gal-H542V achieved a higher GOS yield of 67.08 %, which was the highest in the reported literature. These results suggested that the developed mutagenesis strategy could improve the transgalactosylation efficiency, and the mutant β-gal-H542V could be regarded as a prospective biocatalyst for GOS industrial manufacturing.

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用于高效生物转化半乳寡糖的重组β-半乳糖苷酶，来源于泄殖腔肠杆菌 Zjut HJ2001

半乳寡糖（GOS）作为一种益生元，具有优良的物理和化学特性，可广泛应用于医药和食品领域。近年来，微生物 β-半乳糖苷酶在工业化生产 GOS 方面受到广泛关注。然而，大多数微生物β-半乳糖苷酶的转半乳糖基化活性较低，导致从乳糖酶解转化的 GOS 产率较低。本文从土壤中筛选出了一种全新的β-半乳糖苷酶，并通过丙氨酸扫描和位点饱和突变的组合方法对其进行了成功的过表达、表征和突变。在乳糖浓度为 380 g/L 的条件下，野生型 β-半乳糖苷酶的产率为 51.73%，与之相比，所获得的突变体 β-gal-H542V 的 GOS 产率高达 67.08%，为文献报道的最高产率。这些结果表明，所开发的诱变策略可以提高转半乳糖基化的效率，突变体β-gal-H542V可被视为一种有望用于GOS工业生产的生物催化剂。

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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.