优化酸化球球菌摇摇分批培养条件提高果糖转移酶(左旋蔗糖酶)产量的研究

Ghada Youssef, A. S. Youssef, S. Talha, S. El-Aassar
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引用次数: 3

摘要

微生物果糖转移酶(levansucrase)是参与微生物果糖(levan,菊糖和果寡糖)生物合成的聚合酶。在食品、制药、医药、化妆品、纺织等行业具有潜在的重要性。在结构上,微生物果糖基转移酶蛋白共享糖苷水解酶68家族的催化结构域,并分为7个系统发育相关的簇。这项工作的目的是筛选一些细菌分离物产生细胞外左旋蔗糖酶的效力。从不同来源(口腔拭子、变质牛奶、变质乳清和变质酸奶)中分离出细菌。在摇床条件下发酵2 d后,乳酸球球菌的左旋蔗糖酶活性最高,产量为12.64 U/ml,最大的左旋蔗糖酶产量为15.4 g/L。通过对液体培养液中碳源、氮源、温度和初始pH的优化。事实上,所有碳源都能诱导左旋蔗糖酶的产生,但30% (w/v)的蔗糖是最有效的诱导剂。左旋蔗糖酶的最适温度为30℃,pH为6.0℃。采用选定的沉淀剂对酸碱Pediococcus acilactici提取物的粗左旋蔗糖酶进行富集。结果表明,乙醇(65%饱和度)的纯化系数最高,为3倍,回收率最高,为41.05%。半纯化左旋蔗糖酶的最佳反应温度为40℃,pH为5.2。通过SDS-PAGE对部分纯化的酶进行分析,发现其中一条蛋白带具有左旋蔗糖酶活性,该蛋白带的分子量约为50,000道尔顿。[yousef GA, yousef AS, Talha S, Aassar SA.]优化培养条件提高酸活性Pediococcus acid actitici菌株摇培养中果糖转移酶(左旋蔗糖酶)产量。生命科学学报,2014;11(7):33-47。(ISSN: 1097 - 8135)。http://www.lifesciencesite.com。6
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Study on Increased Fructosyltranseferase (levansucrase) Production by Optimizing Culture Condition from Pediococcus acidilactici Strain in Shaking Batch Cultures
Microbial fructosyltransferases (levansucrases) are polymerases that are involved in microbial fructan (levan, inulin and fructo-oligosaccharide) biosynthesis. Levan has potential importance in food, pharmaceutical, medicine, cosmetics and textile industry. Structurally, microbial fructosyltransferase proteins share the catalytic domain of glycoside hydrolases 68 family and are grouped in seven phylogenetically related clusters. The aim of the work was screening the potency of some bacterial isolates to produce extracellular levansucrase. Bacteria were isolated from different sources (oral swab, spoiled milk, spoiled whey, and spoiled yoghurt). Pediococcus acidilactici showed the highest levansucrase activity after 2 days fermentation under shacking condition yielding (12.64 U/ml), and maximum levan production (15.4 g/L). Following the optimization of carbon source, nitrogen source, temperature and initial pH of the growth medium in submerged liquid cultures. In fact, All carbon sources induced the production of levansucrase activity, but sucrose at 30% (w/v) was the most efficient inducer. The optimal temperature and pH of the levansucrase were 30°C and 6.0, respectively. The crude levansucrase of Pediococcus acidilactici extract was enriched using selected precipitating agents. The results show that ethanol (65% saturation), provided the highest purification factor of 3fold with a recovery yield of 41.05% as a total highest recovered protein. The optimum reaction temperature and pH of semi purified levansucrase were 40°C, 5.2 respectively. Analysis of the partial purified enzyme preparation by SDS–PAGE revealed one protein band showing levansucrase activity, The molecular weight of this band was estimated to be around 50.000 Daltons. [Youssef GA, Youssef AS, Talha S, Aassar SA. Increased Fructosyltranseferase (levansucrase) Production by Optimizing Culture Condition from Pediococcus acidilactici strain in Shaking batchCultures. Life Sci J 2014;11(7):33-47]. (ISSN:1097-8135). http://www.lifesciencesite.com. 6
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