通过合理工程改善菊粉酶的活性,以高效生物合成低分子量菊粉。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-10-03 DOI:10.1007/s00203-024-04153-7
Dawei Ni, Zhaolin Huang, Shuqi Zhang, Yang Yang, Xiaoyong Liu, Wei Xu, Wenli Zhang, Wanmeng Mu
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引用次数: 0

摘要

菊粉是一种被广泛认可的益生元,在各个工业领域都有不同的应用。虽然菊粉主要是通过植物提取生产的,但人们对酶法合成这种替代方法的兴趣日益浓厚。从蔗糖中用酶法生产菊粉,可获得聚合度与植物菊粉相似的聚合物,显示出替代传统提取方法的潜力。本研究从巴达维新杆菌(Neobacillus bataviensis)中发现了一种菊粉蔗糖酶,它展示了一种非加工机制,专门用于合成聚合度从 3 到约 40 的菊粉。该酶在 pH 值为 6.5 和温度为 55 ℃ 时表现出最佳活性,能高效生产菊粉,产量为 50.6%。Ca2+ 可以提高这种酶的活性和耐热性。为了提高催化总活性,研究人员采用了定点突变和截断突变技术,最终确定了一个突变体 T149S,其催化总活性显著提高了 57%。分子动力学模拟揭示了在三个表面区域观察到的灵活性增强对酶活性的积极影响。这项研究不仅为菊粉酶工程学奠定了理论基础,还为菊粉的生产提供了一条潜在的途径。
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Improving the activity of an inulosucrase by rational engineering for the efficient biosynthesis of low-molecular-weight inulin

Inulin, a widely recognized prebiotic, has diverse applications across various industrial sectors. Although inulin is primarily produced through plant extraction, there is growing interest in enzymatic synthesis as an alternative. The enzymatic production of inulin from sucrose, which yields polymers with degrees of polymerization similar to those of plant-derived inulin, shows potential as a viable replacement for traditional extraction methods. In this study, an inulosucrase from Neobacillus bataviensis was identified, demonstrating a non-processive mechanism specifically tailored for synthesizing inulin with polymerization degrees ranging from 3 to approximately 40. The enzyme exhibited optimal activity at pH 6.5 and 55 °C, efficiently producing inulin with a yield of 50.6%. Ca2+ can improve the activity and thermostability of this enzyme. To enhance catalytic total activity, site-directed and truncated mutagenesis techniques were applied, resulting in the identification of a mutant, T149S, displaying a significant 57% increase in catalytic total activity. Molecular dynamics simulations unveiled that the heightened flexibility observed in three surface regions positively influenced enzymatic activity. This study not only contributes to the theoretical foundation for inulosucrase engineering but also presents a potential avenue for the production of inulin.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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