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Transcriptional factors (TFs) play important roles in the responses to oxidative, acid, and other environmental stresses in Gram-positive bacteria, but the regulatory mechanism of TFs involved in oxidative stress remains unknown in lactic acid bacteria. In the present work, homologous overexpression strains with 43 TFs were constructed in the  Lactobacillus plantarum  CAUH2 parent strain. The strain overexpressing CopR displayed the highest sensitivity and a 110-fold decrease in survival rate under H 2 O 2  challenge. The importance of CopR in the response to H 2 O 2  stress was further confirmed by a 10.8-fold increase in the survival of a  copR  insertion mutant.  In silico  analysis of the genes flanking  copR  revealed putative CopR-binding “ cop  box” sequences in the promoter region of the adjacent gene  copB  encoding a Cu 2+ -exporting ATPase. Electrophoretic mobility shift assay (EMSA) analysis demonstrated the specific binding of CopR with  copB in vitro , suggesting  copB  is a target gene of CopR in  L. plantarum . The role of CopB involved in oxidative stress was verified by the significantly decreased survival in the  copB  mutant. Furthermore, a growth defect in copper-containing medium demonstrated that CopB functions as an export ATPase for copper ions. Furthermore, EMSAs revealed that CopR functions as a regulator that negatively regulates  copB  gene and Cu 2+  serves as inducer of CopR to activate the expression of CopB in response to H 2 O 2  stress in  L. plantarum  CAUH2. Our findings indicated that CopR plays an important role in enhancing oxidative resistance by regulating  copB  to modulate copper homeostasis.

The Copper Homeostasis Transcription Factor CopR Is Involved in H2O2 Stress in Lactobacillus plantarum CAUH2