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Glycerol utilization (gyl) operon is relevant to clavulanic acid (CA) biosynthesis in Streptomyces clavuligerus, since its products could take part in supplying glyceraldehyde-3-phosphate (G3P) precursor for CA biosynthesis.The gyl operon is regulated by a member of IclR family regulatory protein, GylR, and is inducible by glycerol.The early and late stages of CA biosynthesis in S.clavuligerus are both positively regulated by CcaR, which is a member of Streptomyces antibiotic regulatory protein (SARP) family and encoded by ccaR gene.In order to improve CA production in S.clavuligerus, an extra copy of ccaRexpressed from Pgyl (the promoter of gyl operon) was integrated into the chromosome of S.clavuligerus NRRL 3585 to coordinate the transcription of CA biosynthesis with glycerol utilization pathway.