Characterization of the anthranilate degradation pathway in Geobacillus thermodenitrificans NG80-2.
Anthranilate is an important intermediate of tryptophan metabolism. In this study, a hydroxylase system consisting of an FADH(2)-utilizing monooxygenase (GTNG_3160) and an FAD reductase (GTNG_3158), as well as a bifunctional riboflavin kinase/FMN adenylyltransferase (GTNG_3159), encoded in the anthranilate degradation gene cluster in Geobacillus thermodenitrificans NG80-2 were functionally characterized in vitro. GTNG_3159 produces FAD to be reduced by GTNG_3158 and the reduced FAD (FADH(2)) is utilized by GTNG_3160 to convert anthranilate to 3-hydroxyanthranilate (3-HAA), which is further degraded to acetyl-CoA through a meta-cleavage pathway also encoded in the gene cluster. Utilization of this pathway for the degradation of anthranilate and tryptophan by NG80-2 under physiological conditions was confirmed by real-time RT-PCR analysis of representative genes. This is believed to be the first time that the degradation pathway of anthranilate via 3-HAA has been characterized in a bacterium. This pathway is likely to play an important role in the survival of G. thermodenitrificans in the oil reservoir conditions from which strain NG80-2 was isolated.
Liu
Xueqian
X
Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.|Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.
Dong
Yangpeng
Y
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Li
Xiaomin
X
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Ren
Yi
Y
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Li
Yanxia
Y
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Wang
Wei
W
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, PR China.|The Engineering and Research Center for Microbial Functional Genomics and Detection Technology, Ministry of Education, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.
Wang
Lei
L
Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.|The Engineering and Research Center for Microbial Functional Genomics and Detection Technology, Ministry of Education, PR China.
Feng
Lu
L
The Engineering and Research Center for Microbial Functional Genomics and Detection Technology, Ministry of Education, PR China.|Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.|The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, PR China.
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005486:Flavin Mononucleotide / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives / Q000378:metabolism; D005798:Genes, Bacterial; D056506:Geobacillus / Q000201:enzymology*; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism*; D053858:Metabolic Networks and Pathways; D005810:Multigene Family; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature; D062367:ortho-Aminobenzoates / Q000378:metabolism*
D001426:Bacterial Proteins; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D005182:Flavin-Adenine Dinucleotide; C058805:1,5-dihydro-FAD; D005486:Flavin Mononucleotide; D010088:Oxidoreductases; D017853:Phosphotransferases (Alcohol Group Acceptor); C022532:riboflavin kinase; D009713:Nucleotidyltransferases; C020143:FMN adenylyltransferase; D006867:Hydrolases