该文在已有的乳酸发酵全合成培养基基础上,研究了氨基酸和维生素类组分对菌体生长和乳酸生成的影响。通过单组分浓度梯度实验确定了对菌体生长和乳酸合成有明显影响的6种氨基酸和3种维生素;以此作为Plackett-Burman实验设计的因素,筛选出了生物素、天冬氨酸、谷氨酸3种对于乳酸生物合成有极显著影响的因素;通过最陡爬坡实验确定3种因素具有最大响应值的浓度范围;并最终通过Box-Behnken实验得出3种因素的最佳添加浓度。在250 mL摇瓶和2.5 L发酵罐中对优化后的培养基进行验证实验,发现优化后合成培养基的乳酸产量、产率和糖酸转化率都较优化前有了显著提高。该实验研究结果为该菌株今后的代谢调控研究奠定了基础。
In this work, the effects of amino acids and vitamins on the growth and lactic acid synthetic metabolism with Lactobacillus paracasei were investigated based on the original chemical defined medium for lactic acid production. Six amino acids and three vitamins which had obvious effects on cell growth and lactic acid production were determined by single component concentration gradient experiments, and they were used for Plackett-Burman experiment getting the result that biotin, aspartic acid and glutamic acid had the most noticeable effects on lactic acid production. Then the range of the three factors that obtained maximum responses value were determined through the steepest experiment, and finally their optimal concentrations were obtained by Box-Behnken experiment. The lactic acid fermentations in shake flask and 2.5 L bioreactor were carried out to verify the above results, showing all the parameters, lactic acid production, productivity and the conversion ratio of lactic acid to glucose, had significant increase compared to the original chemical defined medium. This work paved the way for the quantitative metabolic research of the strain.
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