期刊
ANALYTICAL BIOCHEMISTRY
卷 600, 期 -, 页码 -出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2020.113749
关键词
Fatty acid photodecarboxylase; Flavoprotein; Decarboxylation; Photoinactivation
资金
- Future Biomanufacturing Research Hub - Engineering and Physical Sciences Research Council (EPSRC) [EP/S01778X/1]
- Biotechnology and Biological Sciences Research Council (BBSRC) as part of UK Research and Innovation
- Indian Ministry of Higher Education
- BBSRC [BB/N013980/1]
- BBSRC [BB/N013980/1] Funding Source: UKRI
- EPSRC [EP/S01778X/1] Funding Source: UKRI
Fatty acid photodecarboxylases (FAP) are a recently discovered family of FAD-containing, light-activated enzymes, which convert fatty acids to n-alkanes/alkenes with potential applications in the manufacture of fine and speciality chemicals and fuels. Poor catalytic stability of FAPs is however a major limitation. Here, we describe a methodology to purify catalytically stable and homogeneous samples of recombinant Chlorella variabilis NC64A FAP (CvFAP) from Escherichia coll. We demonstrate however that blue light-exposure, which is required for photodecarboxylase activity, also leads to irreversible inactivation of the enzyme, especially in the absence of palmitate substrate. Photoinactivation is attributed to formation of protein based organic radicals, which were observed by EPR spectroscopy. To suppress photoinactivation, we prepared stable and catalytically active FAP in the dark. The steady-state kinetic parameters of CvFAP (k(cat): 0.31 +/- 0.06 s(-1) and K-M: 98.8 +/- 53.3 mu M) for conversion of palmitic acid to pentadecane were determined using gas chromatography. Methods described here should now enable studies of the catalytic mechanism and exploitation of FAPs in biotechnology.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据