In this study, wood hydrolysate was added in MR medium for the economical production of poly (3-hydroxybutyrate) by recombinant Escherichia coli. When response regulator dr1558 from Deinococcus radiodurans was overexpressed along with PHB biosynthesis genes, recombinant E. coli XB (pKM212_CnCAB+pRadGro_dr1558) had higher cell concentration and PHB content compared to control strain E. coli XB (pKM212_CnCAB+ pRadGro). From the transcriptional analysis by quantitative real time PCR, some genes involved in TCA cycle and PHB biosynthesis were overexpressed in E. coli XB (pKM212_CnCAB+pRadGro_dr1558). This results will be useful for production of biochemicals from cheap hydrolysate by fermentation process.