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四株儿茶酚类铁载体高产菌株产消化酶活性及其益生特性
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山西省优秀人才科技创新项目(201705D211029);山西省重点研发计划项目(201903D221013,201603D221027-3);晋中市科技攻关项目(N1612)


Digestive enzymes and probiotic properties of four bacteria with high yield catechol siderophore
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    摘要:

    【背景】儿茶酚类铁载体对胃肠道菌群的生长代谢具有重要作用,研究儿茶酚类铁载体高产菌株的消化酶活性,挖掘其潜在益生特性具有重要意义。【目的】分析4株分离自健康成人粪样的儿茶酚类铁载体高产菌的产酶特性,通过分析菌株耐酸耐胆盐能力、粘附定殖能力、抗生素耐受性和急性毒性研究其益生特性。【方法】测定4株菌的蛋白酶、淀粉酶、脂肪酶、纤维素酶、植酸酶、乳糖酶和β-葡萄糖苷酶的酶活性。4株菌经人工模拟胃、肠液连续培养后分别计算其活菌数;分析4株菌的自凝集率、黏蛋白粘附率和表面疏水率;对小鼠连续7 d灌胃不同剂量的4株高产菌,观察并记录小鼠的一般体征,计算小鼠脏器指数,进行阳性细菌移位试验。【结果】在试验所测的7种消化酶中,E. coli Gut 07、E. coli Gut 12无蛋白酶和脂肪酶活性,B. cereus Gut 16无乳糖酶活性,E. coli Gut 20无蛋白酶活性,其余均具有。4株菌经人工模拟胃液培养6 h后存活率均大于60%,转移到人工模拟肠液培养24 h后活菌数均大于初始菌落数;该4株菌具备在胃肠道中粘附定殖的能力,对大多数抗生素敏感,在浓度低于4.5×1011 CFU/mL、灌胃剂量为20 mL/kg-bw时对小鼠无急性毒性,无阳性菌株移位现象。【结论】4株儿茶酚类铁载体高产菌株可作为潜在益生菌进行进一步的安全性和功能性研究。

    Abstract:

    [Background] Catechol siderophore play an important role in the growth and metabolism of gastrointestinal flora. [Objective] To study various digestive enzymes of four bacteria producing catechol siderophores isolated from healthy adult feces, and to study their probiotic potential. [Methods] Activities of protease, amylase, lipase, cellulase, phytase, lactase and β-glucosidase were analyzed; The number of live bacteria were counted after culturing four strains in simulated gastric and intestinal juice; and then self-agglutination rate, adhesion rate and surface hydrophobicity of four strains were detected; At lost, for intake safety evaluation different dose of four strains were administrated to mice for 7 days, to observe and record the general signs of mice, to calculate the organ index of mice, and conduct bacterial translocation. [Results] E. coli Gut 07 and E. coli Gut 12 didn’t show protease and lipase activity, B. cereus Gut 16 showed no lactase activity, and E. coli Gut 20 was without protease activity. All the enzymes in this study were detectable in the four selected strains. Regard with the probiotic property, the survival rate of the four strains after 6 hours of simulated gastric juice culture was more than 60%, and the number of live bacteria after 24 hours of simulated intestinal juice culture was greater than the initial colony number. Analysis of self-agglutination rate, adhesion rate and surface hydrophobicity showed that the four strains can adhesion and colonization on the gastrointestinal tract (GIT). For safety evaluation, strains were not tolerant to most of the antibiotics. Intragastric administration (4.5×1011 CFU/mL, 20 mL/kg-bw) didn’t show acute toxicity to mice and no translocation of positive strains. [Conclusion] These four bacteria have the potential as probiotics and need further studying on detail function to human diet and safety assessment after that of acute toxicity.

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武怡荷,胡会玲,陈书明,张希春. 四株儿茶酚类铁载体高产菌株产消化酶活性及其益生特性[J]. 微生物学通报, 2020, 47(6): 1807-1816

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  • 在线发布日期: 2020-06-01
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