拮抗甜瓜枯萎病链霉菌D2菌株发酵条件的优化

Optimition of fermentation conditions of streptomyces strain D2, an antagonistic strepomyces against muskmelon fusarium wilt

  • 摘要: 目的优化甜瓜枯萎病拮抗链霉菌D2菌株的发酵条件,为该菌发酵方法的建立及工业化生产提供科学依据.方法以D2菌株有效活菌数为考察指标,玉米粉含量、豆饼粉含量、pH、摇床转速、接种量和温度为影响因素,利用响应面法对D2菌株的发酵条件进行优化.结果豆饼粉含量、玉米粉含量和温度对D2菌株有效活菌数有显著影响(P<0.05).D2菌株有效活菌数回归模型为:Y=8.28757+0.12344X1+0.21477X2+0.06184X3-0.05929X12-0.09817X22-0.14942X32(式中,Y为有效活菌数对数,X1、X2和X3分别为玉米粉含量、豆饼粉含量和温度的编码值),理论有效活菌数为2.99×108 CFU/mL.D2菌株最优发酵条件为:玉米粉含量39.08 g/L、豆饼粉含量39.19 g/L、培养温度37.4℃,在此条件下D2菌株有效活菌数为3.01×108 CFU/mL,与理论值接近,但极显著高于未经优化采用培养基Ⅱ得到的有效活菌数(1.0×106 CFU/mL)(P<0.01).结论采用响应面法优化得到的D2菌株发酵条件可有效提高该菌株有效活菌数,且回归模型推算出的理论活菌数与优化条件发酵的有效活菌数接近,可用于指导D2菌株的实际规模化生产.

     

    Abstract: ObjectiveThe present study aimed to explore an optimal fermentation condition of streptomyces strain D2 that was against muskmelon fusarium wilt so as to provide scientific basis for the establishment of the strain fermentation method and industrial production.MethodThe viable bacteria number of strain D2 was taken as the indicator of the inves-tigation,and influencing factors included corn powder content,soybean powder content,pH,rotation speed,inoculation amount and temperature. The fermentation condition of strain D2 was optimized by employing the combination of Plackett-Burman(P-B)design,uphill path method,central composite design(CCD)and response surface method.ResultThe re-sults showed that corn powder content,soybean powder content and temperature significantly affected viable bacteria num-ber of strain D2(P<0.05). The regression model of viable bacteria number of strain D2 was:Y=8.28757+0.12344X1+0.21477X2+0.06184X3-0.05929X12-0.09817X22-0.14942X32 ,and the viable bacteria number was 2.99×108 CFU/mL in theo-ry. The optimal fermentation conditions for strain D2 was:corn powder content 39.08 g/L,soybean powder content 39.19 g/L and culture temperature 37.4℃. Under these conditions,the viable bacteria number of strain D2 was 3.01 × 108 CFU/mL, which was close to the theoretical content,but was extremely significant higher than the viable bacteria number(1.0 × 106CFU/mL)obtained by using mediumⅡwithout optimization(P<0.01).ConclusionThe optimum fermentation culture technique for strain D2 optimized by response surface method can effectively improve the viable bacteria number. The via-ble bacteria number predicted by regression model is close to the actual number fermented under the optimized fermenta-tion condition. Therefore,this model can be used to predict the fermentation of D2 strains in practical production.

     

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