Singh Shilpi, Shrivastava Alok Kumar
Arch Microbiol. 2016 Jan;198(1):27-34. doi: 10.1007/s00203-015-1162-8.
The hupL of Anabaena sp. PCC 7120 encodes the large subunit of uptake hydrogenase found in all diazotrophic cyanobacteria and boosts up the nitrogen-fixing potential by catalyzing the removal of the molecular hydrogen produced as a by-product of dinitrogen fixation. Bioinformatics analysis revealed that HupL from Anabaena sp. PCC7120 is a 60.2 kDa, thermostable, glycine-rich protein having highest structural similarity with NiFeSe hydrogenase of Desulfomicrobium baculatumis. Toxicity of selected abiotic stresses like arsenic, cadmium, copper, and salt with HupL was further reconciled by wet-lab approaches like qRT-PCR, hydrogenase and nitrogenase activity assay as hydrogenases unintendedly affect the nitrogenase activity in Anabaena. Down-regulated transcript along with highly inhibited hydrogenase and nitrogenase activities under cadmium stress revealed that cadmium is a potent inhibitor of hydrogenases in Anabaena which indirectly affects its nitrogen-fixing capabilities
鱼腥藻PCC 7120的hupL编码在所有固氮蓝细菌中发现的摄取氢化酶的大亚基,并通过催化去除作为固氮副产物产生的分子氢来提高固氮潜力。生物信息学分析表明,来自鱼腥藻PCC7120的HupL是一种60.2 kDa的热稳定、富含甘氨酸的蛋白质,与杆状脱硫微菌的NiFeSe氢化酶具有最高的结构相似性。通过qRT-PCR、氢化酶和固氮酶活性测定等湿实验室方法,进一步验证了所选非生物胁迫如砷、镉、铜和盐对HupL的毒性,因为氢化酶会意外影响鱼腥藻中的固氮酶活性。镉胁迫下转录本下调以及氢化酶和固氮酶活性受到高度抑制,表明镉是鱼腥藻中氢化酶的有效抑制剂,间接影响其固氮能力。