刚果红法测定乳酸发酵液中的β-葡聚糖类物质  

Determination of β-glucan substances in L-lactic acid fermentation broth by Congo red method

作　　者：韩玉杰[1] 马建卢[1] 寇永磊[1] 樊伟丽[1] 常文耀[1] 吴健[1] 

机构地区：[1]郑州大学生物工程系,郑州450001

出　　处：《食品科技》2011年第10期247-251,255,共6页Food Science and Technology

基　　金：河南省重大公益性科研招标项目(91100910300);郑州大学大学生创新性实验计划项目(2009cxsy039)

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0.025～0.175mol/L,Ca2+浓度为0.001～0.005mol/L,蛋白质质量浓度为0～40mg/L时,这几种杂质对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。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 substance was established to measure the content of β-glucan substance 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 0.025～0.175 mol/L,calcium ions in 0.001～0.005 mol/L,and the concentration of proteins in 0～40 mg/L.The optimal condition for determination by Congo red method is the maximum absorption wavelength 550 nm,pH 8.0 with phosphate buffer and the reaction time 15 minutes.The experiments reveal that the content of β-glucan substance 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.

关 键 词：凝结芽孢杆菌 钙盐法 乳酸 刚果红 Β-葡聚糖 

分 类 号：TS207[轻工技术与工程—食品科学]