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利用来自动物源乳杆菌TMW 1.971的截短型葡聚糖蔗糖酶高产完全线性葡聚糖和异麦芽寡糖。

High-yield production of completely linear dextrans and isomalto-oligosaccharides by a truncated dextransucrase from Ligilactobacillus animalis TMW 1.971.

作者信息

Müller Oliver, Wefers Daniel

机构信息

Institute of Chemistry, Food Chemistry, Martin Luther University Halle-Wittenberg, 06120, Halle (Saale), Germany.

Institute of Chemistry, Food Chemistry, Martin Luther University Halle-Wittenberg, 06120, Halle (Saale), Germany.

出版信息

Carbohydr Res. 2024 Nov;545:109284. doi: 10.1016/j.carres.2024.109284. Epub 2024 Sep 26.

Abstract

Several lactic acid bacteria are capable of producing water-soluble exopolysaccharides such as dextran from sucrose by using glucansucrases. Several recombinant glucansucrases were described, however, yields were often limited and most dextrans were branched at position O3. In this study, the dextransucrase from Ligilactobacillus animalis TMW 1.971 was recombinantly produced without its N-terminal variable region and used for dextran synthesis. The enzyme expressed well and showed very high total as well as transferase activities compared to other glucansucrases. It was able to transfer nearly all glucose from sucrose to oligo- and polymeric products under certain conditions (about 95 % of glucose transferred). The high efficiency of the enzyme made it possible to obtain absolute dextran yields of up to 214.9 g/L from a 1.5 M sucrose solution. Structural characterization of the products showed that the dextrans produced have a rather low molecular weight, a narrow size distribution, and are completely linear. Furthermore, we showed that various low molecular weight dextrans or 1,6-linked isomalto-oligosaccharides can be efficiently produced by acid hydrolysis. Overall, we demonstrated that Ligilactobacillus animalis TMW 1.971 dextransucrase can be used to efficiently synthesize dextrans with a quite unique structural composition. The dextrans produced have a high potential for further applications such as synthesis of copolymers or size standards with a very defined molecular structure.

摘要

几种乳酸菌能够通过使用葡聚糖蔗糖酶从蔗糖中产生水溶性胞外多糖,如右旋糖酐。虽然已经描述了几种重组葡聚糖蔗糖酶,但其产量往往有限,并且大多数右旋糖酐在O3位有分支。在本研究中,重组表达了来自动物源利吉乳酸菌TMW 1.971的葡聚糖蔗糖酶,去除了其N端可变区,并用于右旋糖酐合成。与其他葡聚糖蔗糖酶相比,该酶表达良好,总活性和转移酶活性都非常高。在一定条件下,它能够将几乎所有蔗糖中的葡萄糖转移到寡聚体和聚合物产物中(约95%的葡萄糖被转移)。该酶的高效性使得从1.5M蔗糖溶液中获得高达214.9g/L的绝对右旋糖酐产量成为可能。产物的结构表征表明,所产生的右旋糖酐分子量较低,尺寸分布较窄,并且完全是线性的。此外,我们还表明,通过酸水解可以有效地生产各种低分子量右旋糖酐或1,6-连接的异麦芽寡糖。总体而言,我们证明了动物源利吉乳酸菌TMW 1.971葡聚糖蔗糖酶可用于高效合成具有相当独特结构组成的右旋糖酐。所产生的右旋糖酐在进一步应用方面具有很高的潜力,例如合成具有非常明确分子结构的共聚物或尺寸标准品。

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