土壤分离真菌高效生产曲酸的培养参数优化

Durgadevi Kb, P. Vijayalakshmi, Shilpa, Kumar Bv
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引用次数: 3

摘要

从10种不同的土壤分离真菌中筛选出一株表面发酵产曲酸的新型分离菌。基于18SrRNA的分子分析证实该菌株为黄曲霉FJ537130,为黄曲霉毒素阴性菌株。虽然使用了12种不同的碳源,但西米淀粉的曲酸晶体产量显著。优化条件为底物浓度1000 ml (100 g淀粉粉加入1000 ml H2O中),pH 6.0,时间28 d,温度28℃,蛋白胨浓度4 g/l, KH2PO4浓度1 g/l, MgSO4浓度0.5 g/l,产率为28.5 g/l。所得发酵液经溶剂萃取、凝胶过滤分离曲酸和结晶。通过质子核磁共振、红外光谱和x射线衍射对纯化的曲酸进行了结构表征。采用液相色谱/质谱和高效液相色谱法确定了曲酸的分子量和纯度。分离得到的曲酸晶体对病原菌金黄色葡萄球菌和大肠杆菌具有较高的抑菌活性,最大抑菌区为9 mm。与MDAMB435S(乳腺癌)细胞株相比,曲酸对K562(白血病)细胞株的抑制作用更大。
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Optimization of Cultural Parameters for Cost Effective Production of Kojic Acid by Fungal Species Isolated from Soil
A novel isolate capable of producing opulent yields of kojic acid with surface fermentation was screened from ten different isolated soil fungi. The organism was confirmed as Aspergillus flavus FJ537130 strain using 18SrRNA based molecular analysis and it was identified as a negative producer of aflatoxin. Though twelve different carbon sources were used, significant yields of kojic acid crystals was noticed with Sago starch. The optimized conditions established were substrate concentration 1000 ml (100 g of starch powder in 1000 ml of H2O), pH 6.0, Time 28 d, Temperature 28°C, Peptone concentration 4 g/l, KH2PO4 concentration 1 g/l, MgSO4 concentration 0.5 g/l and the yield achieved was 28.5 g/l. The resulted fermentation broth was subjected to solvent extraction followed by gel filtration for the separation of kojic acid and crystallization. The structural characterization of purified kojic acid was confirmed by Proton NMR, FTIR and XRD. The molecular weight and purity of kojic acid was confirmed by LC/MS and HPLC. The isolated kojic acid crystals shows high antimicrobial activity against pathogenic bacteria Staphylococcus aureus and Escherichia coli and maximum zone of inhibition was 9 mm. The inhibitory effect of kojic acid was more on the cell line K562 (Leukemia) when compared to the MDAMB435S (Breast cancer) cell line.
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