Amaral Danilo T, Oliveira Gabriela, Silva Jaqueline R, Viviani Vadim R
Laboratory of Biochemistry and Biotechnology of Bioluminescence, Graduate Program of Biotechnology and Environmental Monitoring, Department of Chemistry, Physics and Mathematics, Federal University of São Carlos (UFSCar), Campus of Sorocaba, Sorocaba, SP, Brazil.
Photochem Photobiol Sci. 2016 Aug 31;15(9):1148-1154. doi: 10.1039/c6pp00165c. Epub 2016 Jul 25.
Bioluminescent click-beetles display a wide variation of bioluminescence colors ranging from green to orange, including an unusual intra-specific color variation in the Jamaican Pyrophorus plagiophthalamus. Recently, we collected individuals of the Pyrophorus angustus species from the Southern Amazon forest, in Brazil, which displays an orange light emitting abdominal lantern. This species was also previously described from Central America, but displaying a bioluminescence spectrum from 536 nm (dorsal) to 578 nm (ventral). The biogeographic variation of the bioluminescence color in this species could be an adaptation to environmental reflectance and inter/intraspecific sexual competition. Here, we cloned, sequenced, characterized and performed site-direct mutagenesis of this new orange emitting luciferase. The in vitro luciferase spectrum displayed a peak at 594 nm, KM values for ATP and d-luciferin of 160 μM and 17 μM, respectively, and an optimum pH of approximately 8.5. Comparative multialignment and site-directed mutagenesis using different color emitting click-beetle luciferases from P. angustus, Fulgeochlizus bruchi and Pyrearinus termitilluminans luciferases cloned by our group showed an integral role of residue 247 in bioluminescence color modulation.
生物发光叩甲呈现出从绿色到橙色的广泛生物发光颜色变化，包括牙买加的斜斑发光叩甲中一种不寻常的种内颜色变化。最近，我们从巴西南部亚马逊森林采集了窄发光叩甲的个体，其腹部发光器发出橙色光。该物种先前也在中美洲被描述过，但显示出从536纳米（背部）到578纳米（腹部）的生物发光光谱。该物种生物发光颜色的生物地理变化可能是对环境反射率以及种间/种内性竞争的一种适应。在此，我们对这种新的橙色发光荧光素酶进行了克隆、测序、表征并进行了定点诱变。体外荧光素酶光谱在594纳米处有一个峰值，ATP和d - 荧光素的米氏常数分别为160微摩尔和17微摩尔，最佳pH约为8.5。使用我们小组克隆的来自窄发光叩甲、布鲁氏光叩甲和白蚁发光叩甲的不同颜色发光叩甲荧光素酶进行的比较多序列比对和定点诱变表明，第247位残基在生物发光颜色调制中起重要作用。
