通过淀粉样芽孢杆菌莱凡酶和固定化 Ba-SacB 对细胞裂解物的催化作用生产果寡糖糖浆

IF 1.4 4区 生物学 Q3 BIOLOGY Biologia Pub Date : 2024-06-26 DOI:10.1007/s11756-024-01728-z
Song Gao, Xianghui Qi, Ran Lu, Song Zhu, Guoshi Kan, Tatiana Stanishneva-Konovalova, Yingfeng An, Fuping Lu
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引用次数: 0

摘要

利凡诺是一种具有 β-(2, 6) 果糖基连接的果聚糖。近年来,左旋香豆素以其良好的热稳定性、低粘度、假塑性和在水中不溶胀等优异的理化特性,被广泛应用于医药、食品和化工行业。左旋蔗糖酶是负责从底物蔗糖中生物合成左旋香兰的关键酶。本研究以大肠杆菌的细胞裂解液为载体,在多巴胺的作用下对 Ba-SacB(即 Ba-SacB-细胞裂解液)进行固定。与游离的 Ba-SacB 相比,Ba-SacB-细胞裂解物在 pH 值为 6-9 的范围内表现出惊人的稳定性,在 pH 值为 4-5 和 7-10 的范围内相对活性有所提高。在 10%(v/v)丁醇、5mmol/L Fe3+ 的存在下,涡旋 90 分钟后,Ba-SacB-细胞裂解物的相对活性比游离 Ba-SacB 高出 14.32 倍、4.63 倍和 9.2 倍。Ba-SacB 细胞裂解物在重复使用 9 个周期后仍能保持近 81% 的残余活性。通过Ba-SacB-细胞裂解物产生的利凡底物催化利凡酶,高效制备了果寡糖糖浆(FOS),并通过Box-Behnken响应面方法对制备条件进行了优化。
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Production of fructo-oligosaccharide syrup through catalysis of Bacillus amyloliquefaciens levanase and immobilized Ba-SacB on cell lysate

Levan is a type of fructan with β-(2, 6) fructosyl linkage. In recent years, levan has been used in the medicinal, food and chemical industries for its excellent physicochemical properties of good thermal stability, low viscosity, pseudoplasticity, and non-swelling in water. The levansucrase is key enzyme responsible for biosynthesis of levan from substrate sucrose. In the present study cell lysate of Escherichia coli was used as support for convenient immobilization of Ba-SacB (i.e., Ba-SacB-Cell-Lysate) under the action of dopamine. Compared with the free Ba-SacB, the Ba-SacB-Cell-Lysate showed surprisingly enhanced pH stability in the pH range 6–9, and improved relative activities in the pH ranges 4–5 and 7–10. With the presence of 10% (v/v) butyl alcohol, 5mmol/L Fe3+, and after vortex for 90 min, the Ba-SacB-Cell-Lysate showed up to 14.32 times, 4.63 times, and 9.2 times higher relative activities than the free Ba-SacB. The Ba-SacB-Cell-Lysate kept nearly 81% of the residual activity after 9 cycles of reuse. The fructo-oligosaccharide syrup (FOS) was efficiently produced through the catalysis of levanase with the substrate of levan produced by Ba-SacB-Cell-Lysate, and the preparation conditions were optimized by Box-Behnken response surface methodology.

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来源期刊
Biologia
Biologia 生物-生物学
CiteScore
3.30
自引率
6.70%
发文量
290
审稿时长
6 months
期刊介绍: Established in 1946, Biologia publishes high-quality research papers in the fields of microbial, plant and animal sciences. Microbial sciences papers span all aspects of Bacteria, Archaea and microbial Eucarya including biochemistry, cellular and molecular biology, genomics, proteomics and bioinformatics. Plant sciences topics include fundamental research in taxonomy, geobotany, genetics and all fields of experimental botany including cellular, whole-plant and community physiology. Zoology coverage includes animal systematics and taxonomy, morphology, ecology and physiology from cellular to molecular level.
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