Oba Yuichi, Iida Koichiro, Inouye Satoshi
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
FEBS Lett. 2009 Jun 18;583(12):2004-8. doi: 10.1016/j.febslet.2009.05.018. Epub 2009 May 18.
We demonstrated that firefly luciferase has a catalytic function of fatty acyl-CoA synthesis [Oba, Y., Ojika, M. and Inouye, S. (2003) Firefly luciferase is a bifunctional enzyme: ATP-dependent monooxygenase and a long chain fatty acyl-CoA synthetase. FEBS Lett. 540, 251-254] and proposed that the evolutionary origin of beetle luciferase is a fatty acyl-CoA synthetase (FACS) in insect. In this study, we performed the functional conversion of FACS to luciferase by replacing a single amino acid to serine. This serine residue is conserved in luciferases and possibly interacts with luciferin. The mutants of FACSs in non-luminous click beetle Agrypnus binodulus (AbLL) and Drosophila melanogaster (CG6178) gave luminescence enhancement, suggesting that the serine residue is a key substitution responsible for luminescence activity.
我们证明了萤火虫荧光素酶具有脂肪酰辅酶A合成的催化功能[大场洋、小寺真人、井上伸（2003年）。萤火虫荧光素酶是一种双功能酶：ATP依赖性单加氧酶和长链脂肪酰辅酶A合成酶。《欧洲生物化学学会联合会快报》540，251 - 254]，并提出甲虫荧光素酶的进化起源是昆虫中的脂肪酰辅酶A合成酶（FACS）。在本研究中，我们通过将单个氨基酸替换为丝氨酸实现了FACS向荧光素酶的功能转化。该丝氨酸残基在荧光素酶中保守，可能与荧光素相互作用。非发光叩甲Agrypnus binodulus（AbLL）和黑腹果蝇（CG6178）中的FACS突变体发光增强，表明该丝氨酸残基是负责发光活性的关键替代位点。
