Optimization of the Expression Conditions of CGA-N46 in Bacillus subtilis DB1342(p-3N46) by Response Surface Methodology.

摘要: CGA-N46 is a small antifungal derived peptide and consists of the 31st to 76th amino acids N-terminus human chromogranin A. Polycistronic expression recombinant in Bacillus subtilis DB1342 was used improve its production, but yield still low. In present study, response surface methodology (RSM) optimize culture medium composition growth conditions engineered strain B. DB1342(p-3N46) for further increase yield. The results two-level factorial experiments indicated that dextrin tryptone were significant factors affecting expression. Central composite design (CCD) determine ideal each factors. From CCD, optimal predicted be 16.6 g/L, 19.2 KH2PO4·3H2O 6 pH 6.5. And process seed inoculated into fresh at 5% (v/v), followed by 56 hours 30°C induced 2% (v/v) sucrose after one hour shaking culture. To test expression, yeast inhibition assay employed it found under conditions, inhibited C. albican 42.17%, 30.86% more than pre-optimization conditions. summary, RSM can strains DB1342(p-3N46).