Chitosanase-immobilized magnetite-agar gel particles as a highly stable and reusable biocatalyst for enhanced production of physiologically active chitosan oligosaccharides

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology Pub Date : 2024-04-03 DOI:10.1016/j.enzmictec.2024.110443

Takashi Kuroiwa, Yuta Nakagawa, Ryuichi Takayanagi, Akihiko Kanazawa

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Abstract

A novel immobilized chitosanase was developed and utilized to produce chitosan oligosaccharides (COSs) via chitosan hydrolysis. Magnetite-agar gel particles (average particle diameter: 338 μm) were prepared by emulsifying an aqueous agar solution dispersing 200-nm magnetite particles with isooctane containing an emulsifier at 80 °C, followed by cooling the emulsified mixture. The chitosanase from Bacillus pumilus was immobilized on the magnetite-agar gel particles chemically activated by introducing glyoxyl groups with high immobilization yields (>80%), and the observed specific activity of the immobilized chitosanase was 16% of that of the free enzyme. This immobilized chitosanase could be rapidly recovered from aqueous solutions by applying magnetic force. The thermal stability of the immobilized chitosanase improved remarkably compared with that of free chitosanase: the deactivation rate constants at 35 °C of the free and immobilized enzymes were 8.1 × 10⁻⁵ and 3.9 × 10⁻⁸ s⁻¹, respectively. This immobilized chitosanase could be reused for chitosan hydrolysis at 75 °C and pH 5.6, and 80% of its initial activity was maintained even after 10 cycles of use. COSs with a degree of polymerization (DP) of 2–7 were obtained using this immobilized chitosanase, and the product content of physiologically active COSs (DP ≥ 5) reached approximately 50%.

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壳聚糖酶固定化磁铁矿琼脂凝胶颗粒作为一种高度稳定且可重复使用的生物催化剂，用于提高具有生理活性的壳聚糖寡糖的产量

研究人员开发了一种新型固定化壳聚糖酶，并将其用于通过壳聚糖水解生产壳聚糖低聚糖（COS）。磁铁矿-琼脂凝胶颗粒（平均粒径：338 μm）的制备方法是：在 80 °C 下用含有乳化剂的异辛烷乳化分散了 200-nm 磁铁矿颗粒的琼脂水溶液，然后冷却乳化混合物。通过引入乙醛基团进行化学活化，将枯草芽孢杆菌的壳聚糖酶固定在磁铁矿-琼脂凝胶颗粒上，固定化率高（80%），观察到的固定化壳聚糖酶的比活性是游离酶的 16%。这种固定化壳聚糖酶可以通过磁力从水溶液中快速回收。与游离壳聚糖酶相比，固定化壳聚糖酶的热稳定性显著提高：游离酶和固定化酶在 35 °C 时的失活速率常数分别为 8.1 × 10-5 和 3.9 × 10-8 s-1。这种固定化壳聚糖酶可在 75 °C、pH 值为 5.6 的条件下重复用于壳聚糖水解，甚至在使用 10 次后仍能保持 80% 的初始活性。使用这种固定化壳聚糖酶可获得聚合度（DP）为 2-7 的 COS，具有生理活性的 COS（DP ≥ 5）的产物含量达到约 50%。

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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.