Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, P.O. Box 30, 420111 Kazan, Russia.
Int J Mol Sci. 2022 Jul 20;23(14):8009. doi: 10.3390/ijms23148009.
The genome of the neotropical fruit bat was recently sequenced, revealing an unexpected gene encoding a plant-like protein, CYP74C44, which shares ca. 90% sequence identity with the putative CYP74C of . The preparation and properties of the recombinant CYP74C44 are described in the present work. The CYP74C44 enzyme was found to be active against the 13- and 9-hydroperoxides of linoleic and α-linolenic acids (13-HPOD, 13-HPOT, 9-HPOD, and 9-HPOT, respectively), as well as the 15-hydroperoxide of eicosapentaenoic acid (15-HPEPE). All substrates studied were specifically transformed into chain cleavage products that are typical for hydroperoxide lyases (HPLs). The HPL chain cleavage reaction was validated by the identification of NaBH-reduced products (Me/TMS) of 15-HPEPE and 13- and 9-hydroperoxides as (all-)-14-hydroxy-5,8,11-tetradecatrienoic, (9)-12-hydroxy-9-dodecenoic, and 9-hydroxynonanoic acids (Me/TMS), respectively. Thus, CYP74C44 possessed the HPL activity that is typical for the CYP74C subfamily proteins.
最近对新热带水果蝙蝠的基因组进行了测序，揭示了一个意想不到的基因，该基因编码一种类似植物的蛋白质 CYP74C44，其与 的推定 CYP74C 具有约 90%的序列同一性。本工作描述了重组 CYP74C44 的制备和性质。发现 CYP74C44 酶对亚油酸和α-亚麻酸的 13-和 9-过氧化物（分别为 13-HPOD、13-HPOT、9-HPOD 和 9-HPOT）以及二十碳五烯酸的 15-过氧化物（15-HPEPE）具有活性。研究的所有底物都特异性转化为典型的过氧化物裂解酶（HPL）的链裂解产物。通过鉴定 15-HPEPE 和 13-和 9-过氧化物的 NaBH 还原产物（Me/TMS）作为（all-）-14-羟基-5,8,11-十四碳三烯酸、（9）-12-羟基-9-十二烯酸和 9-羟基壬酸（Me/TMS），验证了 HPL 链裂解反应。因此，CYP74C44 具有 CYP74C 亚家族蛋白典型的 HPL 活性。
