Fermentation optimization by response surface methodology for enhanced yield of docosahexaenoic acid by Crypthecodinium cohnii LS1057
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摘要: 为了提高一株隐甲藻藻株LS1057的二十二碳六烯酸产量,对发酵培养基进行了优化。采用Plackett-Burman实验设计法考察发酵培养基中各组分对二十二碳六烯酸产量的影响,结果表明:葡萄糖、酵母膏、KH2PO4的浓度对二十二碳六烯酸的产量影响显著。再用最陡爬坡路径逼近最大响应区域,并结合Box-Behnken实验设计和响应面分析法对3个显著因素进行回归分析,得到优化的发酵培养基组成:葡萄糖79.76g/L、酵母膏14.0g/L、KH2PO40.5g/L、海盐20.0g/L、MgSO4·7H2O 5.0g/L、KNO38.0g/L、FeSO4·7H2O 0.2g/L和M液（M液:V B10.6g/L,V B120.1mg/L）1%（V/V）。采用该法优化培养基,经摇瓶发酵实验供试藻株的二十二碳六烯酸产量达到了1.811g/L,较优化前提高了71.33%。利用70L发酵罐的分批补料发酵实验对优化后的结果做了进一步的验证,发酵结束后二十二碳六烯酸终产量为2.112g/L。实验结果表明经响应面法优化得到的发酵培养基利于隐甲藻LS1057发酵生产二十二碳六烯酸。Abstract: In order to improve the yield of DHA of Crypthecodinium cohnii LS1057, the fermentation medium was optimized.Plackett-Burman design was used to evaluate the effects of medium components on the yield of DHA by C.cohnii LS1057.The results showed that glucose, yeast extract and KH2PO4were the most important factors that influenced the yield of DHA. Based on the experimental results, the path of steepest ascent was undertaken to approach the optimal region of these factors, central composite design and response surface analysis were subsequently employed for further optimization.The optimal medium for C.cohnii LS1057 was obtained, which was composed of 79.76g /L glucose, 14.0g /L yeast extract, 0.5g /L KH2PO4, 20.0g /L sea salt, 5.0g /L MgSO4·7H2O, 8.0g /L KNO3, 0.2g /L FeSO4·7H2O and solution M ( 0.6g /L V B1, 0.1mg /L V B12) 1% ( V/V) . Under the optimum conditions, the maximum yield of DHA reached 1.833g /L in theory and 1.811g /L in the experiment, with an increase of 71.33% compared to the original medium components.The DHA yield of 70 L fermentor reached 2.112g /L under such optimum conditions.The experimental results showed that the conditions optimization by the response surface method were better for enhanced yield of DHA by C.cohnii LS1057.
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