甘薯渣发酵海洋放线菌MA03产生抗黄曲霉毒素有效成分的响应面优化研究
Optimization of fermentation conditions for antiaflatoxin ingredient by marine actinomycete strain MA03 using sweat potato residue with response surface methodology
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摘要: 海洋放线菌菌株MA03能够产生抗黄曲霉毒素有效成分,为了获得其最佳的发酵条件,同时探索甘薯渣资源化利用的新方法,本研究以甘薯渣为营养基质对MA03进行液体发酵,并采用响应面法优化MA03的发酵条件。利用软件Design-Expert 8.0.6通过Plackett-Burman设计,筛选出3个影响产抑制黄曲霉毒素有效成分的显著因素:海水浓度、硫酸铵浓度和接种量,然后经最陡爬坡实验逼近最大响应值区域,再利用Box-Behnken设计对显著因子进行优化。最终确定最优培养条件为:海水浓度0.64%,硫酸铵浓度3.47‰,接种量8.44%,甘薯渣浓度3%,蔗糖浓度2%,硫酸镁浓度1‰,初始p H8.5,发酵温度28℃,发酵时间10d。此条件下黄曲霉毒素抑制率预测值97.62%,对该模型进行验证,验证实验平均值为96.24%±0.38%,与预测值接近,说明该预测模型可靠性较高,可应用于MA03产抗曲霉有效成分的优化。Abstract: Marine actinomycete strain MA03 produced some active ingredient with antiaflatoxin activities. To obtain the optimal fermentation conditions and develop the reutilization of sweet potato residue,the strain MA03 was cultured in the liquid medium supplemented with sweet potato residue. Three main factors(the concentration of sea water,the concentration of(NH4)2SO4and the inoculum size) affected the yield of antiaflatoxin ingredients were screened by Plackett-Burman designed using Design-Expert 8.0.6. Steepest ascent experiments involving these three factors were performed for approaching their optimal response region,and then Box-Behnken design was used to optimize the critical internal factors. Finally,the optimal concentration of the variables were determined as follows:sea water 0.64%,(NH4)2SO43.47‰,inoculum size 8.44%,sweet potato residue 3%,sucrose 2%,Mg SO40.1‰,initial p H 8.5,fermentation temperature 28℃ and fermentation time 10 d. The antiaflatoxin rate was predicted as 97.62% under this condition. This model was confirmed subsequently,and the antiaflatoxin rate was 96.24% ±0.38%,which was close to the predicted value. These results indicated that this predicted model had high reliability,and could be used in optimization of antiaflatoxin ingredient production of marine actinomycete strain MA03.