PCR Amplification and Bioinformatics Analysis of AMO from a Heterotrophic Nitrifier, Alcaligenes faecalis NR
异养硝化菌Alcaligenes faecalis NR氨单加氧酶PCR扩增及生物信息学分析

源生 黄
2017 Advances in Microbiology  
Ammonia monooxygenase (AMO) is one of the key enzymes related to nitrification process, and the corresponding gene is amo. In this study, two pairs of primers were designed according to the amo gene sequence in the NCBI database. The amo gene of Alcaligenes faecalis strain NR was amplified successfully, and 1639 bp length DNA sequence was obtained. This sequence contained a complete open reading frame (ORF), which encodes a 346 aa length polypeptide chain. BLAST analysis results showed that
more » ... lts showed that this polypeptide was AMO protein sequences. This protein belonged to hydrophobic and basic protein, and the molecular weight was 36.7 kDa. In addition, AMO was transmembrane protein which contained 12 transmembrane regions. Tertiary structure model of AMO was built successfully by using threading method, and the confidence of this model was 98.6%. AMO contained a nickel atom binding sites. The conformation was good according to Ramachandran plot. This investigation offers reference for further understanding the key enzymes relating to heterotrophic nitrification.
doi:10.12677/amb.2017.62006 fatcat:li6prq3c4nbcpmclmlu7hymo4y