发酵液中植酸酶活性测定的方法的研究.docVIP

发酵液中植酸酶活性测定方法的研究 摘 要：以钒钼法对发酵液中植酸酶活性进行了测定，并对影响酶活性测定结果的因素进行了研究。实验结果表明，钒钼法测磷的灵敏度和线性范围分别为0.025mmol/L和0.025mmol/L~0.60mmol/L；为减小植酸酶活性的测定误差，结果测定必须在颜色反应结束后 的4h以内完成。研究中同时发现，发酵液含有的无机磷远远大于酶活测定产生的无机磷含量，直接用发酵液测得的植酸酶酶活是不可靠的。通过对透析2h的发酵液进行植酸酶酶活的测定，实验结果的最大相对偏差仅为7.1%。 关 键 词：钒钼法；植酸酶；透析；酶活 Study on analysis methods of phytase activity in fermentation broth Abstract: Determination of phytase activity in fermentation broth by vanadiu-molybdenum method and the factors influenced results were studied. The results showed that the sensitivity and linear range for inorganic phosphorus by vanadiu-molybdenum method were 0.025 mmol/L and 0.025-0.60 mmol/L, respectively. Detection must be finished within 4 h after colour reaction for decrease of error. It was also found that concentration of inorganic phosphorus in fermentation broth was much higher than that of produced by enzyme activity and determination of phytase activity directly with fermentation broth was not reliable. The relative standard deviation of the results was 7.1% when the fermentation broth was dialysed for 2 h. Key words: vanadiu-molybdenum method; phytase; dialysis; phytase activity 收稿日期：2009-12-24 基金项目：泰山医学院青年科学基金项目（ 2008-2009 ） 作者简介：洒荣波（ 1978- ），男，讲师，研究方向为微生物发酵 。 phenotypes and receptor structure [J]. Yeast，2004，7(6)：559-574. [9] FUJII T, HORIC Y. Further investigation on a substance isolated from wort inducing early flocculation [J]. Rep Res Lab Kirin Brew Co, 1975,18:75-85. [10] FUJINO S, YOSHIDA T. Premature flocculation of yeast induced by some wort constituents [J]. Rep Res Lab Kirin Brew Co,1976,19:45. !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! 分析与检测 ·1 67·2010 No．4 Serial No．217 China Brewing 植酸酶活力测定的准确性是保证植酸酶研究和生产 顺利的基础，关于植酸酶测定方法的报道很多，主要采用比 色法，但其一直存在着误差大、不能准确定量的问题 [1-4] 。 目 前测定植酸酶活力应用较广的有 4种方法：钼酸铵-FeSO 4 法 、钼酸铵-VC法、钼酸铵-氨基奈酚磺酸法以及钒钼法。其 中钒钼法于1994年被美国分析化学专家列入AOAC（Associ- ation of Analytical Communities）标准方法，我国也于2002 年将钒钼法作为测定饲用植酸酶活性的国家标准 [5] 。 本研究优化所得到的植酸酶发酵培养基配方中含有 3g/L 的磷酸盐 [6] ，磷酸盐的作用不仅是提供酵母生长所需 的