Cloning and characterization of a NADP+-malic enzyme gene from Bradyrhizobium japonicum USDA110

• Published in: Soil science and plant nutrition (Tokyo) Vol. 48; no. 5; pp. 711 - 717
• Main Authors: Sinsuwongwat, S. (Chiangmai Univ. (Thailand)), Kodera, A, Kaneko, T, Tabata, S, Nomura, M, Tajima, S
• Format: Journal Article
• Language: English
• Published: Tokyo Taylor & Francis Group 01.10.2002; Japanese Society of Soil Science and Plant Nutrition
• Subjects: Biological and medical sciences; BRADYRHIZOBIUM JAPONICUM; dicarboxylic acid; Fundamental and applied biological sciences. Psychology; gene cloning; MALIC ENZYME; malic enzymes; MOLECULAR CLONING; NADP; NITROGEN FIXATION; symbiotic nitrogen fixation; symbiotic nitrogen fixation; gene cloning; C4-dicarboxylic acid; NADP+-malic enzymes; Bradyrhizobium japonicum
• ISSN: 0038-0768; 1747-0765
• DOI: 10.1080/00380768.2002.10409261

Malic enzymes have been considered to play a key role in energy metabolism for nitrogenase reaction in bacteroids. To elucidate the physiological role of the malic enzymes in Bradyrhizobium japonicum bacteroids, a putative malic enzyme gene Bjtme1 was cloned by polymerase chain reaction (PCR) using degenerated primers from conserved regions of the protein sequences of bacterial malic enzymes and draft sequence data of the Bradyrhizobium japonicum USDA110 genome sequence project. To confirm the characteristics of the Bjtme1 gene, the protein encoded by this gene was over-expressed using a pET32a(+) system and it exhibited a NADP + -malic enzyme (EC 1.1.1.40) activity, indicating that Bjtme1 was the gene of the NADP + -malic enzyme. This is the first report on the cloning and characterization of the NADP + -malic enzyme gene from B. japonicum, and the gene structure was compared with that of NADP + -malic enzyme genes of other rhizobia.