解淀粉芽孢杆菌LOR10耐热植酸酶的生产及生化特性研究

Arastoo Badoei-dalfard, Maryam Parhamfar
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摘要

植酸酶可以通过在胃和食品加工行业中的植酸酶消化来提高蛋白质的消化率和矿物质的可利用性,从而提高植物性食品的营养价值。微生物来源更有希望在商业规模上生产植酸酶。本研究的目的是从具有商业价值的温泉中筛选和分离植酸酶产生菌。利用16S rDNA基因对最佳分离株进行了分子鉴定。在不同的磷酸盐、氮和碳源存在下,制备了植酸酶生产的优化。在不同pH、温度和离子化合物的存在下,还探索了酶的活性和稳定性。使用Clustal omega将分离物LOR10的16S rDNA基因序列与GenBank中的序列进行比较,显示与解淀粉芽孢杆菌具有98%的序列同源性。培养基优化研究表明,半乳糖、酵母提取物和磷酸三钙分别是生产植酸酶的最佳碳源、氮源和磷酸盐源。还观察到最适温度活性为70℃。植酸酶的稳定性在5.0–8.0的pH范围内达到最佳。在CaCl2、ZnCl2和MnSO4存在下,植酸酶活性分别提高了1.4、2.3和1.6倍。可以指出,在EDTA和SDS存在下,植酸酶活性降低了约30%。基于这些结果,可以得出结论,LOR10植酸酶作为动植物性食品的添加剂具有巨大的商业潜力。
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Production and biochemical characterization of a thermostable phytase from Bacillus amyloliquefaciens LOR10
. Phytase can improve the nutritional value of plant-based foods by enhancing protein digestibility and mineral availability through phytate digestion in the stomach and the food processing industry. Microbial sources are more promising for the production of phytases on a commercial scale. The objectives of this exploration were to screening and isolation of phytase-producing bacteria from hot spring with commercial interest. Molecular identification of the best isolate was achieved by the 16S rDNA gene. Optimization of phytase production was prepared in the presence of different phosphate, nitrogen, and carbon sources. Enzyme activity and stability were also explored in the presence of different pHs, temperatures, and ion compounds. Comparing the 16S rDNA gene sequence of the isolate LOR10 with those in GenBank using Clustal omega shows 98% sequence homology with Bacillus amyloliquefaciens. Medium optimization studies showed that galactose, yeast extract, and tricalcium phosphate were the best sources of carbon, nitrogen, and phosphate for phytase production, respectively. The optimum temperature activity was also observed to be 70 o C. Phytase stability was at its optimum in a pH range of 5.0–8.0. Phytase activity increased in the presence of CaCl 2 , ZnCl 2 , and MnSO 4 about 1.4, 2.3 and 1.6 folds, respectively. It could be mentioned that phytase activity decreased by about 30 % in the presence of EDTA and SDS. On the basis of the results, it could be concluded that LOR10 phytase has a great potential for commercial interest as an additive to animal plant-based foods.
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