过氧化氢酶高产菌株EIM-70的鉴定及产酶培养基的优化

Identification of Catalase-producing Strain EIM-70 and Its Optimization of Submerged Fermentation Mediums

[目的]对过氧化氢酶高产菌株EIM-70进行鉴定,并对其产酶培养基进行优化。[方法]对EIM-70菌株进行形态学分析及16SrDNA序列的系统发育进化分析,以鉴定其所属菌种;在采用单因素试验确定EIM-70产过氧化氢酶的最适碳、氮源基础上,采用Plackett-Burman试验设计筛选出对产酶影响显著的因子,再利用最陡爬坡试验和响应面分析法确定最适产酶培养基配方。[结果]试验将EIM-70菌株鉴定为枯草芽孢杆菌(Bacillus subitilis);EIM-70的最适产酶培养基为:葡萄糖19.25 g/L、NaNO39.25 g/L、FeSO4.7H2O2.46×10-3g/L、MgSO4.7H2O 0.50 g/L、Na2HPO4.12H2O 9.25 g/L、KH2PO40.60 g/L、2°麦芽汁,优化后Bacillus subitilis EIM-70的液体发酵产过氧化氢酶的活力可达6 927.45 U/ml,比优化前提高1.87倍。[结论]该研究为过氧化氢酶的工业化生产奠定了基础。

[ Objective] The aim was to identify a high catalase-producing strain EIM-70 and optimize its submerged fermentation mediums. [ Method] The strain EIM-70 was identified by the morphological analysis and phylogenetic analysis of 16S rDNA sequence; the optimal carbon and nitrogen sources for EIM-70 on catalase production were obtained by single-factor experiments, the Plackett - Burman design was adopted to screen out the significant related factors based on this, the optimal fermentation medium was obtained by the steepest ascent search experi- ment and response surface methodology. [ Result ] The strain EIM-70 was identified as Bacillus subitilis; the optimized medium for eatalase production was glucose 19.25 g/L, NaNOs 9.25 g/L, FeSO4 · 7H20 2.46 × 10 -s g/L, MgSQ4 · 7H20 0.50 g/L, N%HPO4 · 12H20 9.25 g/L, KH2PO4 0.60 g/L and 2°malt extract, in this medium, the catalase activity of Bacillus subitilis EIM-70 was up to 6 927.45 U/ml and 1.87 times higher than control. [ Conclusion] It provided foundation for the industrlalized nroduotirm rff cntalase.