Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany) http://biotech.uni-greifswald.de.
Angew Chem Int Ed Engl. 2014 Jan 27;53(5):1439-42. doi: 10.1002/anie.201306952. Epub 2013 Dec 20.
Flavonoids are a large group of plant secondary metabolites with a variety of biological properties and are therefore of interest to many scientists, as they can lead to industrially interesting intermediates. The anaerobic gut bacterium Eubacterium ramulus can catabolize flavonoids, but until now, the pathway has not been experimentally confirmed. In the present work, a chalcone isomerase (CHI) and an enoate reductase (ERED) could be identified through whole genome sequencing and gene motif search. These two enzymes were successfully cloned and expressed in Escherichia coli in their active form, even under aerobic conditions. The catabolic pathway of E. ramulus was confirmed by biotransformations of flavanones into dihydrochalcones. The engineered E. coli strain that expresses both enzymes was used for the conversion of several flavanones, underlining the applicability of this biocatalytic cascade reaction.
类黄酮是一大类植物次生代谢物,具有多种生物特性,因此引起了许多科学家的兴趣,因为它们可以产生具有工业应用价值的中间体。厌氧肠道细菌真杆菌(Eubacterium ramulus)可以分解类黄酮,但到目前为止,该途径尚未通过实验得到证实。在本研究中,通过全基因组测序和基因基序搜索,鉴定出一种查尔酮异构酶(CHI)和烯醇还原酶(ERED)。这两种酶可以在大肠杆菌中以其活性形式成功克隆和表达,甚至在有氧条件下也可以。通过将黄烷酮生物转化为二氢查尔酮,证实了真杆菌的分解途径。表达这两种酶的工程大肠杆菌菌株被用于几种黄烷酮的转化,突出了这种生物催化级联反应的适用性。