比色法测定细脚拟青霉胞内甘露醇及羟自由基清除率  

Determination of Intracellular Mannitol Content and Maximum Scavenging Rate of Hydroxyl Radical of Paecilomyces tenuipes by Colorimetry

作　　者：桂琳[1] 葛飞[2] 黄寅[2] 

机构地区：[1]皖南医学院微生物与免疫学教研室,安徽芜湖241000 [2]安徽工程科技学院生物化学工程系,安徽芜湖241000

出　　处：《安徽农业科学》2009年第19期8818-8819,8824,共3页Journal of Anhui Agricultural Sciences

基　　金：皖南医学院中青年科研基金(WK200906);安徽高校省级自然科学研究项目(KJ2009B071);安徽工程科技学院引进人才科研启动基金(2007YQ005);芜湖市科技计划项目(2008505)

摘　　要：[目的]采用比色法测定细脚拟青霉菌丝体中的甘露醇含量及羟自由基清除率。[方法]以细脚拟青霉为供试菌株，用超声提取法提取细脚拟青霉菌丝体中的甘露醇，以D-甘露醇为标准品，绘制其标准曲线并计算其回归方程，通过测定甘露醇的吸光度计算细脚拟青霉菌丝体中的甘露醇含量及羟自由基清除率。同。时，测定培养时间和提取时间对菌丝体中甘露醇含量和抗氧化活性的影响。[结果]甘露醇在412nm处有最大吸收峰，质量浓度在10—80μg／ml范围内其具有良好的线性关系，其回归方程为：C=103．09A-1．3814，r=0．9988；平均回收率为101．47％，RSD为2．6％（n=5）。菌丝体中的甘露醇含量及羟自由基清除率的变化趋势基本不随培养时间改变，培养第6天时均达到最高值，分别为73．56μg/ml和47．95％。[结论]为进一步研究细脚拟青霉的抗氧化功能提供理论依据。[ Objective] The purpose was to determine the mannitol and scavenging rate of hydroxyl radical in fermented mycilia of Paecilomyces tenuipes by using colofimetry metheod. [ Method ] With P. tenuipes as tested strains, the mannitol was extracted by using ultrasonic extraction from the mycelia of P. tenuipes . With D-mannitol as the standard substance, the standard curve was drew and the regression equation was calculated. The mannitol content and scavenging rate of hydroxyl radicals were calculated by measuring the absorbanee of mannitol.. At the same time, the culture time and extraction time were determined to study their effect on the mannitol content and scavenging rate of hydroxyl radical in myeelium of P. tenuipes. [ Result] Mannitol had the maximum absorption peak at the 412 nm and its linear relationship was better when the content of mannitol was 10 - 80 μg/m]. The regression equation was C = 103.09A - 1. 381 4, r = 0. 998 8, and the average recovery was 101.47% , RSD =2.6% （n = 5 ）. The trends of the mannitol content and scavenging rate of hydroxyl radical in mycilia did not change with the culture time, and they could reach as highest as 73.56μg/ml and 47.95% resp. after culture for 6 d. [ Conclusion] This study provided a theoretical basis for further study on the antioxidant function of P. tenuipes.

关 键 词：比色法 细脚拟青霉 甘露醇 羟自由基清除率 

分 类 号：Q936[生物学—微生物学]