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生理聚集的乳糖酶以循环批次模式应用于海藻糖半乳糖基化反应。

Physiologically Aggregated LacZ Applied in Trehalose Galactosylation in a Recycled Batch Mode.

作者信息

Belkova Martina, Janegova Tatiana, Hrabarova Eva, Nahalka Jozef

机构信息

Institute of Chemistry, Centre for Glycomics, Slovak Academy of Sciences, Dubravska Cesta 9, SK-84538 Bratislava, Slovakia.

Institute of Chemistry, Centre of Excellence for White-Green Biotechnology, Slovak Academy of Sciences, Trieda Andreja Hlinku 2, SK-94976 Nitra, Slovakia.

出版信息

Life (Basel). 2023 Jul 25;13(8):1619. doi: 10.3390/life13081619.

Abstract

Galactooligosaccharides obtained via β-galactosidase transgalactosylation have health-promoting properties and are widely recognized as effective prebiotics. Trehalose-based galactooligosaccharides could be introduced into food and pharmaceutical industries similarly to trehalose. In light of this, new technological approaches are needed. Recently, in vivo enzyme immobilizations for recombinant proteins have been introduced, and physiological aggregation into active inclusion bodies (aIBs) has emerged as one such method of in vivo immobilization. To prepare LacZ β-galactosidase in the form of aIBs, we used a short 10 amino acid aggregation-prone tag. These native protein particles were simply washed from the cell lysate and applied in trehalose galactosylation in a recycled batch mode. In this study, aIBs entrapped in alginate beads, encapsulated in alginate/cellulose sulfate/poly(methylene-co-guanidine) capsules and magnetized were compared with free aIBs. Alginate/cellulose sulfate/PMCG capsules showed more suitable properties and applicability for biotransformation of trehalose at its high concentration (25%, /) and elevated temperature (50 °C).

摘要

通过β-半乳糖苷酶转半乳糖基化获得的低聚半乳糖具有促进健康的特性,被广泛认为是有效的益生元。基于海藻糖的低聚半乳糖可以像海藻糖一样引入食品和制药行业。有鉴于此,需要新的技术方法。最近,已经引入了用于重组蛋白的体内酶固定化技术,生理聚集形成活性包涵体(aIBs)就是其中一种体内固定化方法。为了制备aIBs形式的乳糖酶β-半乳糖苷酶,我们使用了一个由10个氨基酸组成的易于聚集的短标签。这些天然蛋白质颗粒只需从细胞裂解物中洗涤出来,并以循环分批模式用于海藻糖半乳糖基化反应。在本研究中,将包埋在藻酸盐珠中、封装在藻酸盐/硫酸纤维素/聚(亚甲基-co-胍)胶囊中并进行磁化的aIBs与游离aIBs进行了比较。藻酸盐/硫酸纤维素/聚甲基胍胶囊在高浓度(25%,/)海藻糖和高温(50°C)下对海藻糖的生物转化表现出更合适的特性和适用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa96/10455999/d6c0a24af860/life-13-01619-g001.jpg

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