刚果红法测定乳酸发酵液中的β-葡聚糖类物质 精灵论文.doc

刚果红法测定乳酸发酵液中的 β-葡聚糖类 物质 常文耀，王莎莎，汤小兵，王文方，韩玉杰，樊伟丽，吴建 （郑州大学生物工程系，郑州 450001） 5 摘要：在利用凝结芽孢杆菌（Bacillus coagulans）进行以葡萄糖为基质的乳酸发酵过程中 通过外加商品纤维素酶可以显著增加乳酸产量。为探讨其中原因，用羧甲基纤维素（CMC） 作为β-葡聚糖标品，建立了刚果红分光光度法测定乳酸发酵提取液中β-葡聚糖的稳定体系, 用以测定乳酸发酵液中的β-葡聚糖含量。实验证明当 Na+浓度为 0.025mol/L-0.125mol/L， Ca2+浓度为 0.001 mol/L-0.005mol/L，蛋白质质量浓度为 0mg/L-40mg/L 时，这几种杂质对 10 β-葡聚糖测定的干扰可以忽略。刚果红法测定β-葡聚糖的最佳条件为：最大吸收波长 550nm、pH8.0 磷酸缓冲液、测定时间 15min。实验结果显示乳酸发酵液中β-葡聚糖含量很 低，纤维素酶水解发酵液中的β-葡聚糖类物质不足于显著增加乳酸产量，其增产作用一定 另有原因。 关键词：凝结芽孢杆菌；钙盐法；乳酸；刚果红；β-葡聚糖 15 中图分类号：TQ016.1 Determination of β-glucan substances in L-lactic acid fermentation broth by Congo red method CHANG Wenyao, WANG Shasha, TANG Xiaobing, WANG Wenfang, HAN Yujie, 20 FAN Weili, WU Jian (Department of Bioengineering, Zhengzhou University, ZhengZhou 450001) Abstract: In the process of L-lactic acid fermentation by Bacillus coagulans using glucose as the feedstock, the yield of L-lactic acid could be increased through adding cellulases. To investigate the reason, a stable system of spectrophotometric determination of β-glucan substances was established to 25 measure the content of β-glucan substances in the L-lactic acid fermentation broth, using carboxymethyl cellulose (CMC) as the standard substance of β-glucan. It was illustrated that the potential influence might be omitted when the concentration of sodium ions in 0.025mol/L-0.175mol/L, calcium ions in 0.001 mol/L-0.005mol/L, and the concentration of proteins in 0mg/L-40mg/L. The optimal conditions for determination by Congo red method are the maximum absorption wavelength 30 550nm, pH 8.0 with phosphate buffer and the reaction time 15 minutes. The experiments reveal that the content of β-glucan substances in L-lactic acid fermentation broth is rather low, and the reason why cellulases could enhance the yield of L-lactic acid needs further investigation. Keywords:Bacillus coagulans; calcium lactate; L-lactic acid; Congo red; β-glucan substances 35 0 引言 乳酸是一种重要的有机酸，在食品、化工、医药、纺织等领域应用广泛[1,2,3]。尤