Technology Research Laboratory, Shimadzu Corporation, Kyoto 619-0237, Japan.
Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama 226-8503, Japan.
ACS Chem Biol. 2022 Apr 15;17(4):864-872. doi: 10.1021/acschembio.1c00897. Epub 2022 Mar 16.
Luciferases are widely used as sensitive reporters in various fields ranging from basic biology to medical diagnosis, public health, and food inspection. Scientists have isolated novel luciferases from bioluminescent organisms and concentrated on improving their brightness and thermostability. Recently, small bright luciferases such as artificial luciferase (ALuc) (21 kDa), NanoLuc (19 kDa), GLuc (18 kDa), and TurboLuc (16 kDa) have been reported. However, smaller, brighter, and more stable luciferases are desired for further applications. Here, we constructed the smallest and bright mutant of ALuc, named "picALuc" (13 kDa). picALuc retained the luminescence activity of the full-length ALuc; moreover, its brightness and thermostability were at the same levels as NanoLuc. Furthermore, we showed the advantage of picALuc for the bioluminescence resonance energy transfer-based assay due to its smallness. Our development has opened the door for wider and more practical applications of luciferases.
荧光素酶被广泛用作从基础生物学到医学诊断、公共卫生和食品检验等各个领域的敏感报告基因。科学家们已经从生物发光生物体中分离出新型荧光素酶，并专注于提高其亮度和热稳定性。最近，已经报道了一些小型明亮的荧光素酶，如人工荧光素酶 (ALuc) (21 kDa)、NanoLuc (19 kDa)、GLuc (18 kDa) 和 TurboLuc (16 kDa)。然而，为了进一步应用，人们希望获得更小、更亮、更稳定的荧光素酶。在这里，我们构建了最小和最亮的 ALuc 突变体，命名为“picALuc”(13 kDa)。picALuc 保留了全长 ALuc 的发光活性；此外，其亮度和热稳定性与 NanoLuc 相当。此外，我们由于其小巧的尺寸，展示了 picALuc 在基于生物发光共振能量转移的测定中的优势。我们的开发为荧光素酶的更广泛和更实际的应用开辟了道路。
