Carrier immobilization and auto-exposition favoring reuse of silyletherase SilE-R from Brassica sp. with high activity and enantiospecificity.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-05-29 DOI:10.1007/s10529-025-03600-9

Lisa Pick, Anna L Schumacher, Elif Öztel, Thorsten Mascher, Marion B Ansorge-Schumacher

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引用次数: 0

Abstract

Objectives: Investigation of immobilization methods promoting the use of silyletherases from Brassica sp. for the efficient and enantiospecific hydrolysis of silyl-protected hydroxyl functions.

Results: Different supports for adsorptive and covalent binding of the silyletherase SilE-R as well as exposure of the enzyme on the surface of Bacillus subtilis endospores, so-called SporoBeads, were evaluated. While the highest protein loading of 26 mg enzyme per gram was obtained by adsorptive binding, the best combination of specific activity and enantiospecificity was obtained when SilE-R was exposed on SporoBeads. Protein loading was estimated at 2.6 mg per gram of spore, which was in the same range as after covalent binding to a carrier. In six repeated reaction cycles, SporoBeads exposing SilE-R lost less than 10% of their catalytic activity. The enantiomeric excess could not be increased even with short reaction times, but remained constant over all repeated cycles.

Conclusion: The exposure of silyletherases on SporoBeads has been identified as a promising approach for the synthetic application of this novel type of enzyme, although some properties relevant for catalytic applications need to be further improved.

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具有高活性和对映体特异性的芸苔属硅醚化酶SilE-R的载体固定化和自体暴露。

目的：研究利用芸苔属植物的水飞蓟醚化酶对映体特异性水解水飞蓟保护羟基功能的固定化方法。结果：评价了不同载体对水飞蓟醚酶SilE-R的吸附和共价结合作用，以及该酶在枯草芽孢杆菌内生孢子（SporoBeads）表面的暴露。通过吸附结合获得的蛋白载量最高，为26 mg / g酶，而将SilE-R暴露在SporoBeads上获得了最佳的特异性活性和对映体特异性组合。蛋白质负荷估计为每克孢子2.6毫克，这与与载体共价结合后的蛋白质负荷范围相同。在六个重复的反应循环中，暴露于sil - r的SporoBeads的催化活性损失不到10%。对映体的过量即使在较短的反应时间内也不会增加，但在所有重复循环中都保持不变。结论：在SporoBeads上暴露水飞蓟醚酶是一种很有前途的合成方法，但在催化应用方面仍需进一步改进。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.