Identification and Optimized Fermentation of High Catalase-producing Serratia marcescens, FZSF02
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摘要: 为解决过氧化氢废水对环境的污染，从稻田土中筛选到1株高产过氧化氢酶菌株。经形态学观察和分子生物学鉴定，确认该菌属于黏质沙雷氏菌，命名为Serratia marcescens FZSF02。通过培养基碳源、氮源、无机盐类型及产过氧化-氢酶条件优化。在培养条件为：1%大豆蛋白胨、0.5%麦芽糖、0.2% NaCl、31℃、180 r·min-1、发酵时间48 h，菌株产过氧化氢酶总酶活最高达6 380 U·mL-1，比优化前提高了7.4倍。粗酶液最适反应温度为60℃，最适反应pH为8.0。该菌在过氧化氢废水无害化处理和其他工农业过程中有很好的应用潜力。Abstract: To search for a high catalase-producing bacteria strain, field samples and lab collections were screened. FZSF02 was initially selected and subjected to morphological examinations and 16S rRNA sequencing.It was identified to be a strain of Serratia marcescens. Subsequently, various carbon and nitrogen sources, mineral salts, and fermentation conditions to maximize the catalase production of the bacteria were experimented and optimized. It was found that a medium consisting of 1% soy peptone, 0.5% lactose, and 0.2% NaCl applied for the fermentation at 31℃ with a constant shaking at 180rpm for 48 h could yield the enzyme at the highest level of 6 380 U·mL-1 among all tested conditions. The yield was 7.4 times of what was without the optimization. The peak activity of the obtained crude catalase was determined to reach at 60℃ and pH 8.0.The enzyme could potentially be applied for treating hydrogen peroxide pollutionin waste water effluent.
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Key words:
- Serratia marcescens /
- catalase /
- fermentation conditions
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图 2 菌株FZSF02与其近缘菌株的16SrDNA系统发育树
Figure 2. Phylogenetic tree of FZSF02 based on sequence comparability of 16S rDNA
图 3 氮源对菌株FZSF02过氧化氢酶产量的影响
Figure 3. Effect of nitrogen sources on catalase production of FZSF02
图 4 碳源对菌株FZSF02过氧化氢酶产量的影响
Figure 4. Effect of carbon sources on catalase production of FZSF02
图 5 发酵温度和摇床转速对过菌株FZSF02产过氧化氢酶的影响
Figure 5. Effect of fermentation temperature and shaker speed on catalase production
图 6 发酵时间对菌株FZSF02产过氧化氢酶的影响
Figure 6. Effect of fermentation duration on catalase production
图 7 菌株FZSF02所产过氧化氢酶的最适反应温度和pH值
Figure 7. Optimal temperature and pH for peak activity of catalase from FZSF02
表 1 无机盐对菌株FZSF02过氧化氢酶产量的影响
Table 1. Effect of mineral salts on catalase production of FZSF02
无机盐
/0.2%过氧化氢酶活力
/(U·mL-1)NaCl 4 385.3±537 a Na2HPO4 2 724.8±83 d FeCl3 0 CuSO4 0 NH4NO3 1 045.9±83 g NaH2PO4 1 770.6±47 f KCl 3 156±126 c MnSO4 2 743.1±171 d CH3COONa 1 972.5±95 ef CaCl2 2 229.4±82 e (NH4)2SO4 2 880.7±95 cd MgSO4 1 752.3±169 f 对照组 3 568±223 b 
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表 2 正交试验因素水平
Table 2. Factors and levels of orthogonal experiment
水平 A大豆蛋白胨
/(g·L-1)B麦芽糖
/(g·L-1)C NaCl
/(g·L-1)1 5 5 1 2 10 10 2 3 15 15 3
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表 3 正交试验结果
Table 3. Results of orthogonal experiment
试验号 A B C 酶活/
(g·L-1)1 1 1 1 2324 2 1 2 2 2438 3 1 3 3 2248 4 2 1 2 2420 5 2 2 3 1426 6 2 3 1 3936 7 3 1 3 1822 8 3 2 1 3216 9 3 3 2 2574 K1 7010 6566 9476 K2 7782 7080 7432 K3 7612 8758 5496 k1 2337 2189 3159 k2 2594 2360 2477 k3 2537 2919 1832 R 257 730 1327
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