Identification and characterization of a novel thermostable PL7 alginate lyase from a submarine volcanic metagenomic library

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Enzyme and Microbial Technology Pub Date : 2024-07-21 DOI:10.1016/j.enzmictec.2024.110486

Vasileios Tsopanakis , Elena Anastasiadou , Maria D. Mikkelsen , Anne S. Meyer , Ioannis V. Pavlidis

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Abstract

Seaweed biomass is as an abundant and renewable source of complex polysaccharides, including alginate which has a variety of applications. A sustainable method for exploiting alginate towards the production of valuable oligosaccharides is through enzymatic processing, using alginate lyases. Industrial refinement methods demand robust enzymes. Metagenomic libraries from extreme environments are a new source of unique enzymes with great industrial potential. Herein we report the identification of a new thermostable alginate lyase with only 58 % identity to known sequences, identified by mining a metagenomic library obtained from the hydrothermal vents of the volcano Kolumbo in the Aegean Sea (Kolumbo Alginate Lyase, KAlLy). Sequence analysis and biochemical characterization of KAlLy showed that this new alginate lyase is a Polysaccharide Lyase of family 7 (PL7) enzyme with endo- and exo-action on alginate and poly-mannuronic acid, with high activity at 60°C (56 ± 8 U/mg) and high thermostability (half-life time of 30 h at 50°C). The response surface methodology analysis revealed that the reaction optimum conditions with poly-mannuronic acid as substrate are 44°C, pH of 5.5 with 440 mM NaCl. This novel alginate lyase is a valuable addition to the toolbox of alginate modifying enzymes, due to its diverse sequence and its good thermal stability.

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从海底火山元基因组文库中鉴定新型恒温 PL7 藻酸盐裂解酶并确定其特征

海藻生物质是复杂多糖（包括具有多种用途的海藻酸）的丰富且可再生的来源。利用藻酸盐生产有价值的低聚糖的可持续方法是使用藻酸盐裂解酶进行酶处理。工业精炼方法需要强健的酶。来自极端环境的元基因组文库是具有巨大工业潜力的独特酶的新来源。在本文中，我们报告了通过挖掘从爱琴海科伦坡火山热液喷口获得的元基因组文库（科伦坡藻酸盐酶，KAlLy）发现的一种新的恒温藻酸盐裂解酶，其与已知序列的同一性仅为 58%。对 KAlLy 的序列分析和生化鉴定表明，这种新的藻酸盐裂解酶属于多糖裂解酶家族 7（PL7），对藻酸盐和聚甘露糖酸具有内切和外切作用，在 60°C 时具有高活性（56 ± 8 U/mg ）和高热稳定性（50°C 时半衰期为 30 小时）。响应面方法分析表明，以聚甘露糖酸为底物的最佳反应条件为 44°C、pH 5.5 和 440 mM NaCl。这种新型藻酸盐裂解酶的序列多样，热稳定性好，是藻酸盐改性酶工具箱中的宝贵补充。

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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.