Department of Chemical and Environmental Engineering, University of California, Riverside, California.
Department of Bioengineering, University of California, Riverside, California.
Biotechnol Bioeng. 2018 Mar;115(3):557-564. doi: 10.1002/bit.26492. Epub 2017 Nov 22.
We report a spore-based biocatalysis platform capable of producing and self-assembling active multimeric enzymes on a spore surface with a high loading density. This was achieved by co-expressing both a spore surface-anchoring scaffoldin protein containing multiple cohesin domains and a dockerin-tagged enzyme of interest in the mother cell compartment during Bacillus subtilis sporulation. Using this method, tetrameric β-galactosidase was successfully displayed on the spore surface with a loading density of 1.4 × 10 active enzymes per spore particle. The resulting spore biocatalysts exhibited high conversion rates of transgalactosylation in water/organic emulsions. With easy manufacture, enhanced thermostability, excellent reusability, and long-term storage stability at ambient temperature, this approach holds a great potential in a wide range of biocatalysis applications especially involving organic phases.
我们报告了一种基于孢子的生物催化平台，能够在孢子表面以高装载密度生产和自组装具有高活性的多聚体酶。这是通过在枯草芽孢杆菌孢子形成期间在母细胞室中共同表达含有多个黏合结构域的孢子表面锚定支架蛋白和带有 dockerin 标签的目标酶来实现的。使用这种方法，四聚体β-半乳糖苷酶成功地在孢子表面展示，每个孢子颗粒的装载密度为 1.4×10 个活性酶。所得的孢子生物催化剂在水/有机乳液中具有高的转糖苷转化率。这种方法具有易于制造、增强的热稳定性、优异的可重复使用性和在环境温度下的长期储存稳定性，在广泛的生物催化应用中具有很大的潜力，特别是涉及有机相的应用。
