Valency engineering of monomeric enzymes for self-assembling biocatalytic hydrogels
Valency engineering of monomeric enzymes for self-assembling biocatalytic hydrogels
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DOI：
10.1039/c9sc04074a
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发表时间：
2019-11-14
期刊：
影响因子：
8.4
通讯作者：
Niemeyer, Christof M.
中科院分区：
文献类型：
--
作者：
Bitterwolf, Patrick;Gallus, Sabrina;Niemeyer, Christof M.
全酶水凝胶是连续流动生物催化的有效试剂。这些材料可以通过两种寡聚酶的自组装获得，并用互补的SpyTag和SpyCatcher单元进行修饰。为了便于获得大量具有生物催化作用的单体酶，我们证明了来自酿酒酵母的单体(S)立体选择性酮还原酶Gre2p的标记价可以设计成与枯草芽孢杆菌的辅因子再生葡萄糖1-脱氢酶GdH组装稳定、有活性的水凝胶。这些凝胶安装在微流控反应器中，在连续流动超过8天的情况下，对前手性甲基酮的还原显示出高的转化率和立体选择性。流动反应器模块的顺序使用和‘向上编号’的并行化表明，这种方法适合于半制备规模的合成。
All-enzyme hydrogels are efficient reagents for continuous flow biocatalysis. These materials can be obtained by self-assembly of two oligomeric enzymes, modified with the complementary SpyTag and SpyCatcher units. To facilitate access to the large proportion of biocatalytically relevant monomeric enzymes, we demonstrate that the tagging valency of the monomeric (S)-stereoselective ketoreductase Gre2p from Saccharomyces cerevisiae can be designed to assemble stable, active hydrogels with the cofactor-regenerating glucose 1-dehydrogenase GDH from Bacillus subtilis. Mounted in microfluidic reactors, these gels revealed high conversion rates and stereoselectivity in the reduction of prochiral methylketones under continuous flow for more than 8 days. The sequential use as well as parallelization by 'numbering up' of the flow reactor modules demonstrate that this approach is suitable for syntheses on the semipreparative scale.