应用高效液相色谱分析藏灵菇源乳酸菌发酵液中的四碳二羧酸

Analysis of the C4-Dicarboxylatic Acid in Broth Produced by Lactic acid bacteria from Tibetan Kefir using HPLC

目的:分析藏灵菇源乳酸菌发酵液中的四碳二羧酸,研究发酵条件对合成四碳二羧酸的影响。方法:采用高效液相色谱分析藏灵菇源乳酸菌Tx、KS4、KL1菌株发酵液中的四碳二羧酸,通过四因素三水平[L9(34)]正交试验确定高产四碳二羧酸的发酵条件。结果:发酵液中草酰乙酸含量0.03318 g/L,延胡索酸含量0.05002 g/L,苹果酸含量0.000046 g/L;优化的发酵条件是:乳酸菌Tx、KS4与KL1菌种比例为3:3:3,接种量2%,发酵温度40℃,发酵时间24 h。在此条件下,合成四碳二羧酸含量是优化前的27.25倍,其中,草酰乙酸含量是优化前的41.21倍,延胡索酸含量是优化前的17.08倍,苹果酸含量是优化前的1 001.3倍。结论:在优化的发酵条件下,藏灵菇源乳酸菌混合发酵液中的四碳二羧酸含量明显提高。

Objective：The C4-Dicarboxylatic Acid,produced by Lactic acid bacteria from Tibetan Kefir,was analyzed by HPLC.The effect of fermentation condition on the synthesis of C4-Dicarboxylatic Acid was investigated.Methods：The C4-Dicarboxylatic Acid in broth,and produced by strains Tx,KS4 and KL1 of Lactic acid bacteria from Tibetan Kefir was analyzed by HPLC.The optimizing mixed culture fermentation condition of C4-Dicarboxylatic Acid was determined by four factors and three levels orthogonal experiments [L9（34）].Results：The content of oxaloacetic acid fumarate and malic acid in broth were 0.03318 g/L,0.05002 g/L,and 0.000046 g/L respectively.The optimal fermentation conditions were as follows：The ratio of strains Tx,KS4 and KL1 was 3：3：3,and inoculum was 2%,and fermentation temperature was 40 ℃,and fermentation time was 24 h.Under the optimal fermentation conditions,the content of C4-Dicarboxylatic Acid was 27.25 times higher than before.Among the total,the content of oxaloacetic acid fumarate and malic acid were 41.21,17.08 and 1 001.3 times respectively higher than before.Conclusions：The content of C4-Dicarboxylatic Acid in broth of Lactic acid bacteria from Tibetan Kefir,was obviously increased,and provided the further research foundation.