Enhancing stability and transglycosylation efficiency of alternansucrase from Leuconostoc citreum ABK-1 by immobilization

IF 3.4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biocatalysis and agricultural biotechnology Pub Date : 2025-07-01 DOI:10.1016/j.bcab.2025.103671

Kusuma Thongplaew , Karan Wangpaiboon , Pawinee Panpetch , Rath Pichyangkura , Thanapon Charoenwongpaiboon , Manchumas Prousoontorn

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Abstract

Alternansucrase (ALT) catalyzes the transglycosylation of sucrose, synthesizing α-1,6 and α-1,3-linked glucan, and also utilizes maltose as an acceptor to produce glucooligosaccharides. In this study, alternansucrase from Leuconostoc citreum ABK-1 was immobilized on glutaraldehyde-activated chitosan beads. Immobilization parameters, namely glutaraldehyde concentration and enzyme loading, were optimized to achieve 59.7 % activity yield with an immobilized activity of 10.3 U/g bead. The immobilized ALT exhibited a broader working pH than that of free enzyme, with an optimal pH shift from 4 to 5–6. The thermostability of immobilized ALT was significantly improved, exhibiting up to a 5.4-fold increase in half-life at 37 °C. DLS analysis revealed that the immobilized ALT produced larger alternan particles, with an average diameter of 117.3 nm compared to 79.75 nm for the free enzyme, suggesting that transglycosylation efficiency was enhanced. Also, the immobilized ALT could synthesize a higher amount of isomaltose and isomaltotriose. Moreover, we found that the immobilized enzyme retained over 50 % of its initial activity after 14 reuse cycles, suggesting its potential for industrial applications of immobilized ALT due to enhanced stability and reusability.

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固定化法提高柠檬酸白菌ABK-1交替蔗糖酶的稳定性和转糖基化效率

交替蔗糖酶（ALT）催化蔗糖的转糖基化，合成α-1,6和α-1,3连接的葡聚糖，也利用麦芽糖作为受体生产低聚糖。本研究以戊二醛活化的壳聚糖珠为载体，固定化了柠檬酸白菌ABK-1的交替蔗糖酶。通过优化戊二醛浓度和酶载量，固定化酶活率为59.7%，固定化酶活为10.3 U/g球。与游离酶相比，固定化ALT的工作pH值更宽，最佳pH值为4 ~ 5 ~ 6。固定化ALT的热稳定性得到了显著改善，在37℃时的半衰期增加了5.4倍。DLS分析表明，固定的ALT产生了更大的交替颗粒，平均直径为117.3 nm，而自由酶的平均直径为79.75 nm，表明转糖基化效率提高。同时，固定化后的ALT可以合成更多的异麦芽糖和异麦芽糖。此外，我们发现固定化酶在重复使用14次后仍保持了50%以上的初始活性，这表明固定化ALT具有增强的稳定性和可重复使用性，具有工业应用潜力。

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来源期刊

Biocatalysis and agricultural biotechnology Agricultural and Biological Sciences-Agronomy and Crop Science

CiteScore

7.70

自引率

2.50%

发文量

308

审稿时长

48 days

期刊介绍： Biocatalysis and Agricultural Biotechnology is the official journal of the International Society of Biocatalysis and Agricultural Biotechnology (ISBAB). The journal publishes high quality articles especially in the science and technology of biocatalysis, bioprocesses, agricultural biotechnology, biomedical biotechnology, and, if appropriate, from other related areas of biotechnology. The journal will publish peer-reviewed basic and applied research papers, authoritative reviews, and feature articles. The scope of the journal encompasses the research, industrial, and commercial aspects of biotechnology, including the areas of: biocatalysis; bioprocesses; food and agriculture; genetic engineering; molecular biology; healthcare and pharmaceuticals; biofuels; genomics; nanotechnology; environment and biodiversity; and bioremediation.