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Biosynthesis of Low Molecular Weight Antifungal Protein from Aspergillus giganteus in Batch Fermentation and In-Vitro Assay

机译:在分批发酵和体外测定中的曲霉菌黄霉菌低分子量抗真菌蛋白的生物合成

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摘要

In present study, Taguchi's design of experiment L-9 orthogonal array was created using Qualitek-4 software with four most critical factors namely, K2HPO4, MgSO4, CaCl2 and culture pH. Production of a new intracellular antifungal protein in submerged fermentation was optimized with yield of 0.98 +/- 0.1 mg/gram dry cell weight mycelia from Aspergillus giganteus MTCC 8408. The average molecular mass of the purified protein was figured as 5.122 kDa using Electro Spray Ionization-Mass Spectrometry. Scanning electron microscopy was used to correlate the effect of selected factors on fungal cell morphology and its metabolite production. In vitro antifungal susceptibility assay was profiled against Aspergillus niger and minimum inhibitory concentrations were in the range 0.3 +/- 0.06 mu g/ml. The stronger influencing factors on afp production and mycelial biomass were noted with CaCl2 and K2HPO4 respectively. The validation experiments using optimized conditions confirmed an improvement in afp by 3.86 times with mycelial biomass by 1.52 times, compared to the basal medium. The present statistical optimization study revealed an opportunity to promote economical design at the industrial level for future scale up of effective antifungal agent against systemic aspergillosis as well as possible post harvest loss.
机译:在目前的研究中,Taguchi的实验L-9正交阵列的设计是使用Qualitek-4软件创建具有四种最关键因素的QualityK-4软件,即K2HPO4,MgSO 4,CaCl2和培养pH。从Aspergillus Giganteus MTCC 8408的产率为0.98 +/- 0.1mg /克干细胞重量菌丝体,优化了浸没式发酵中的新细胞内抗真菌蛋白质。使用电光电离为5.122kDa,纯化蛋白的平均分子量为5.122kDa -质谱。扫描电子显微镜用于将选定因子对真菌细胞形态及其代谢产物产生的影响相关。体外抗真菌敏感性测定患者对Aspergillus尼日尔进行分析,并且最小抑制浓度为0.3 +/-0.06μg/ ml。 CaCl2和K2HPO4分别注意到AFP生产和菌丝体生物质上的强烈影响因素。使用优化条件的验证实验证实了AFP的改善3.86次,与基础培养基相比,菌丝体生物质的生物量为1.52倍。目前的统计优化研究揭示了促进工业水平经济设计的机会,以便未来扩大有效的抗真菌剂免受全身性曲柄的抑制以及可能的收获后损失。

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