ACIB GmbH, Graz, Austria.
Institute of Chemistry, University of Graz, NAWI Graz, BioTechMed Graz, Graz, Austria.
Appl Microbiol Biotechnol. 2018 Jul;102(14):6057-6068. doi: 10.1007/s00253-018-9052-z. Epub 2018 May 12.
The formation of C-C bonds by using CoA independent acyltransferases may have significant impact for novel methods for biotechnology. We report the identification of Pseudomonas strains with CoA-independent acyltransferase activity as well as the heterologous expression of the enzyme in E. coli. The cloning strategies and selected expression studies are discussed. The recombinant acyltransferases were characterized with regard to thermal and storage stability, pH,- and co-solvent tolerance. Moreover, the impact of bivalent metals, inhibitors, and other additives was tested. Careful selection of expression and working conditions led to obtain recombinant acyltransferase form Pseudomonas protegens with up to 11 U mL activity.
通过使用 CoA 非依赖性酰基转移酶形成 C-C 键，可能对生物技术的新方法产生重大影响。我们报告了具有 CoA 非依赖性酰基转移酶活性的假单胞菌菌株的鉴定，以及该酶在大肠杆菌中的异源表达。讨论了克隆策略和选择表达研究。研究了重组酰基转移酶的热稳定性、储存稳定性、pH 值和共溶剂耐受性。此外，还测试了二价金属、抑制剂和其他添加剂的影响。仔细选择表达和工作条件，可使恶臭假单胞菌来源的重组酰基转移酶的活性最高达到 11 U mL。
