Department of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Germany.
Enzyme Microb Technol. 2013 Sep 10;53(4):283-7. doi: 10.1016/j.enzmictec.2013.01.007. Epub 2013 Jan 25.
In order to establish a new route for ɛ-caprolactone production from the corresponding cyclohexanol with an internal cofactor recycling for NADPH, a recently redesigned thermostable polyol dehydrogenase (PDH) and the cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus were combined. First, the expression of PDH could be improved 4.9-fold using E. coli C41 with co-expression of chaperones. Both enzymes were also successfully co-immobilized on glutaraldehyde-activated support (Relizyme™ HA403). Cyclohexanol could be converted to ɛ-caprolactone (ɛ-CL) with 83% conversion using the free enzymes and with 34% conversion using the co-immobilized catalysts. Additionally, a preparative scale biotransformation of ɛ-caprolactone starting from cyclohexanol was performed using the soluble enzymes. The ɛ-CL could be isolated by simple extraction and evaporation with a yield of 55% and a purity of >99%.
为了从相应的环己醇建立一条新的ε-己内酯生产路线，并对 NADPH 进行内部辅助因子循环，将一种最近重新设计的耐热多元醇脱氢酶（PDH）和来自醋酸钙不动杆菌的环己酮单加氧酶（CHMO）结合在一起。首先，通过共表达伴侣，大肠杆菌 C41 可以将 PDH 的表达提高 4.9 倍。这两种酶也可以成功地共固定在戊二醛活化的载体（Relizyme™ HA403）上。环己醇可以在游离酶的作用下转化为 83%的ε-己内酯（ε-CL），在共固定化催化剂的作用下转化为 34%的ε-CL。此外，使用可溶性酶在制备规模上从环己醇开始进行了ε-己内酯的生物转化。ε-CL 可以通过简单的萃取和蒸发来分离，产率为 55%，纯度大于 99%。
