Pick Lisa, Schumacher Anna L, Öztel Elif, Mascher Thorsten, Ansorge-Schumacher Marion B
Chair of Molecular Biotechnology, Dresden University of Technology, Dresden, Germany.
Chair of General Microbiology, Dresden University of Technology, Dresden, Germany.
Biotechnol Lett. 2025 May 29;47(3):59. doi: 10.1007/s10529-025-03600-9.
Investigation of immobilization methods promoting the use of silyletherases from Brassica sp. for the efficient and enantiospecific hydrolysis of silyl-protected hydroxyl functions.
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.
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.
研究能促进使用来自芸苔属植物的甲硅烷基醚酶，以高效且对映体特异性水解甲硅烷基保护的羟基官能团的固定化方法。
评估了用于甲硅烷基醚酶SilE-R吸附和共价结合的不同载体，以及该酶在枯草芽孢杆菌芽孢表面（即所谓的芽孢珠）的暴露情况。通过吸附结合获得了每克最高26毫克酶的蛋白质负载量，而当SilE-R暴露在芽孢珠上时，获得了比活性和对映体特异性的最佳组合。估计每克芽孢的蛋白质负载量为2.6毫克，这与共价结合到载体后的范围相同。在六个重复反应循环中，暴露SilE-R的芽孢珠失去的催化活性不到10%。即使反应时间很短，对映体过量也无法提高，但在所有重复循环中保持恒定。
已确定将甲硅烷基醚酶暴露在芽孢珠上是这种新型酶合成应用的一种有前景的方法，尽管一些与催化应用相关的性质需要进一步改进。
