Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Org Biomol Chem. 2011 Oct 7;9(19):6727-33. doi: 10.1039/c1ob05565h. Epub 2011 Aug 12.
The oxofunctionalization of saturated hydrocarbons is an important goal in basic and applied chemistry. Biocatalysts like cytochrome P450 enzymes can introduce oxygen into a wide variety of molecules in a very selective manner, which can be used for the synthesis of fine and bulk chemicals. Cytochrome P450 enzymes from the CYP153A subfamily have been described as alkane hydroxylases with high terminal regioselectivity. Here we report the product yields resulting from C(5)-C(12) alkane and alcohol oxidation catalyzed by CYP153A enzymes from Mycobacterium marinum (CYP153A16) and Polaromonas sp. (CYP153A P. sp.). For all reactions, byproduct formation is described in detail. Following cloning and expression in Escherichia coli, the activity of the purified monooxygenases was reconstituted with putidaredoxin (CamA) and putidaredoxin reductase (CamB). Although both enzyme systems yielded primary alcohols and α,ω-alkanediols, each one displayed a different oxidation pattern towards alkanes. For CYP153A P. sp. a predominant ω-hydroxylation activity was observed, while CYP153A16 possessed the ability to catalyze both ω-hydroxylation and α,ω-dihydroxylation reactions.
饱和烃的氧化官能化是基础和应用化学中的一个重要目标。细胞色素 P450 酶等生物催化剂可以非常选择性地将氧引入各种分子中，可用于精细化学品和大宗化学品的合成。属于 CYP153A 亚家族的细胞色素 P450 酶已被描述为具有高末端区域选择性的烷烃羟化酶。在这里，我们报告了分枝杆菌（CYP153A16）和极地单胞菌（CYP153A P. sp.）来源的 CYP153A 酶催化 C(5)-C(12)烷烃和醇氧化的产物产率。对于所有反应，详细描述了副产物的形成。在大肠埃希氏菌中克隆和表达后，用假单胞菌 putidaredoxin（CamA）和 putidaredoxin reductase（CamB）重新构成了单加氧酶的活性。尽管这两种酶系统都产生了伯醇和α,ω-烷二醇，但它们对烷烃的氧化模式不同。对于 CYP153A P. sp.，观察到主要的ω-羟化活性，而 CYP153A16 则具有催化ω-羟化和α,ω-二羟化反应的能力。
