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In this study, the gentisate and protocatechuate pathways in  Burkholderia xenovorans  LB400 were analyzed by genomic and functional approaches, and their role in 3-hydroxybenzoate (3-HBA) and 4-hydroxybenzoate (4-HBA) degradation was proposed. The LB400 genome possesses two identical  mhbRTDHI  gene clusters encoding the gentisate pathway and one  mhbM  gene encoding a 3-HBA 6-hydroxylase that converts 3-HBA into gentisate. The  pca  genes encoding the protocatechuate pathway and the  pobA  gene encoding the 4-HBA 3-monooxygenase that oxidizes 4-HBA into protocatechuate are arranged in gene clusters and single genes mainly at the minor chromosome, but also at the major chromosome and the megaplasmid. Strain LB400 was able to grow on gentisate, protocatechuate, 3-HBA and 4-HBA. Transcriptional analyses showed that the  mhbD  gene encoding the gentisate 1,2-dioxygenase was expressed during growth on 3-HBA, 4-HBA and gentisate, whereas the  pcaG  gene encoding the protocatechuate 3,4-dioxygenase was expressed only during growth on 4-HBA and protocatechuate. The  mhbM  gene encoding the 3-HBA 6-hydroxylase was transcribed in strain LB400 during growth on HBAs, gentisate, protocatechuate and glucose. The  pobA  gene encoding the 4-HBA 3-monooxygenase was expressed during growth on HBAs and glucose. 3-HBA- and 4-HBA-grown LB400 cells showed gentisate 1,2-dioxygenase activity, whereas protocatechuate 3,4-dioxygenase activity was observed only in 4-HBA-grown cells. The  mhbR  gene encoding a MarR-type transcriptional regulator that probably regulates the expression of the MhbT transporter, and the  pcaQ  and  pcaR  genes encoding LysR-type transcriptional regulators that regulate  pcaHG  and  pcaIJBDC  genes, respectively, were transcribed during growth on both HBAs, gentisate, protocatechuate and glucose, suggesting a basal constitutive expression. The results indicate active gentisate, protocatechuate, 3-HBA and 4-HBA catabolic pathways in  B. xenovorans  LB400 and suggest that 3-HBA is channeled exclusively through the gentisate route, whereas 4-HBA is funneled into the protocatechuate central pathway and potentially into the gentisate pathway.

plos one

Genomic and Functional Analyses of the Gentisate and Protocatechuate Ring-Cleavage Pathways and Related 3-Hydroxybenzoate and 4-Hydroxybenzoate Peripheral Pathways in Burkholderia xenovorans LB400