摘要
出芽短梗霉(Aureobasidium pullulans)发酵产聚苹果酸(PMA)的研究近年来受到广泛关注。发酵液中聚苹果酸的测定通常是将聚苹果酸水解成苹果酸后,通过分光光度法(Goodban法)或HPLC法进行测定。本文先对乙醇沉淀法测定聚苹果酸含量进行了研究,发现乙醇沉淀法无法将聚苹果酸选择性地沉淀出来,沉淀物中含有一定量的普鲁兰多糖。因此进一步建立了先乙醇沉淀再分光光度法测定的新方法,即:将乙醇沉淀物重新溶解,经酸水解后再以分光光度法进行测定。试验结果表明,该方法与HPLC法的检测结果比较接近,而且可以免除发酵液中黑色素产生对分光光度法测定的干扰,可以对出芽短梗霉发酵液中的聚苹果酸含量进行准确测定。
The research of fermentative production of poly-malic acid(PMA)by Aureobasidium pullulans has attracted extensive attentions in recent years. The determination of PMA was usually by measuring the amount of malic acid by spectrophotometry or HPLC after hydrolysis. The precipitation of PMA from fermentation broth by ethanol was studied. The results showed that PMA could not be precipitated selectively by ethanol,and certain amount of pullulan was detected in the precipitate. A new method was established by combining precipitation of PMA and spectrophotometry,the process was as follows :after ethanol precipitation,the perecipitate was dissolved in pure water and hydrolysis by H2 SO4,the obtained hydrolysate was used for malic acid detection by spectrophotometry. The study results were very similar to that of HPLC,so the new method could be used for accurate measurement of PMA in Aureobasidium pullulans fermentation broth without concerning of black pigments interfering by spectrophotometer.
引文
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