Tailoring of levansucrase product size by a comparative molecular dynamics approach.

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology Pub Date : 2024-12-23 DOI:10.1016/j.enzmictec.2024.110577

Zhiwei Li, Tong Bao, Kaiwen Chen, Chao Hu, Xinyu Zhang, Xueqin Hu, Jingwen Yang, Hongbin Zhang

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引用次数: 0

Abstract

Levan is widely used as food additives. Its utilization is significantly influenced by its molecular weight. Bacillus subtilis levansucrase (Bs-SacB) and Priestia megaterium levansucrase (Pm-SacB) yield levan of different weights. To delve deeper into the molecular underpinnings of the molecular weight disparity between the products of these two enzymes, we conducted a focused study on the eight loops surrounding the active sites of Bs-SacB and Pm-SacB and identified Loop3 and loop4 as critical determinants in changing the molecular weight of Bs-SacB 's products. Subsequently, leveraging mutation energy analysis and non-homologous substitution strategies, we crafted tailored modifications in loop3 and loop4, yielding a spectrum of mutant enzymes that exhibit diverse molecular weight profiles including F182Y (3698 Da), CYTI (3093 Da), 3-Pbl (2776 Da), 4-Bml (1845 Da), and F182K (1571 Da). This research provide a novel comparative molecular dynamics approach to change product molecular weight and it is successfully applied in the modification of levansucrase.

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.