title: Multiple pqqA genes respond differently to environment and one contributes dominantly to pyrroloquinoline quinone synthesis.
abstract: Pyrroloquinoline quinone is the third redox cofactor after nicotinamide and flavin in bacteria, and its biosynthesis pathway comprise five steps initiated from a precursor peptide PqqA coded by pqqA gene. Methylovorus sp. MP688 is equipped with five copies of pqqA genes. Herein, the transcription of pqqA genes under different conditions by real-time quantitative PCR and β-galactosidase reporter genes are reported. Multiple pqqA genes were proved to play significant roles and contribute differently in PQQ synthesis. pqqA1, pqqA2, and pqqA4 were determined to be dominantly transcribed over the others, and correspondingly absence of any of the three genes caused a decrease in PQQ synthesis. Notably, pqqA was up-regulated in low pH and limited oxygen environment, and it is pqqA2 promoter that could be induced when bacteria were transferred from pH 7.0 to pH 5.5. Deletion analysis revealed a region within pqqA2 promoter inhibiting transcription. PQQ concentration was increased by overexpression of pqq genes under control of truncated pqqA2 promoter. The results not only imply there exist negative transcriptional regulators for pqqA2 but also provide us a new approach to achieve higher PQQ production by deleting the target binding sequence.
authors.affiliation: Laboratory of Microorganism Engineering, Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Ge
authors.forename: Wenxi
authors.initials: W
authors.lastname: Wang
authors.forename: Baohua
authors.initials: B
authors.lastname: Du
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
authors.forename: Xianghua
authors.initials: X
authors.lastname: Xiong
authors.forename: Weicai
authors.initials: W
authors.lastname: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D042762:Methylophilaceae / Q000235:genetics* / Q000378:metabolism; D005810:Multigene Family; D009154:Mutation; D045542:PQQ Cofactor / Q000096:biosynthesis* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D045542:PQQ Cofactor
keywords: PQQ production; Promoter; Pyrroloquinoline quinine; Transcriptional regulation; pqqA