Optimization of fermentation medium for spore production of Paenibacillus polymyxa IN937a and its antifungal activity

Pham Trung Hieu, Tran Dai Lam, Le The Tam, Vu Thi Thoa, Nguyen Thi Phuong Thao, Ho Tu Cuong, Vu Dinh Hoang, Le Dang Quang
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Abstract

In this study, we optimized the fermentation medium for spore production of Paenibacillus polymyxa IN937a. Seven factors including molasses, glucose, magnesium sulfate, potassium pyrophosphate, yeast extract, zinc sulfate, and ammonium sulfate are selected as the basis for the screening of factors affecting the spore production of P. polymyxa IN937a by the Plackett-Burman experiment. Based on the analysis of the Plackett-Burman matrix, the result showed that yeast extract, molasses, and ammonium sulfate were the three main impact factors (P < 0.05), which affected the yield of P. polymyxa IN937a spores. Then, the optimum combination of the three factors was subsequently optimized by the response surface methodology (RSM) using Box-Behnken design to increase the spore production in P. polymyxa IN937a fermentation. The obtained results by RSM predicted that maximum spore density of P. polymyxa IN937a was 6.606×109 spore/mL after 48 hours of the experiment when the appropriate medium for the spore production of P. polymyxa IN937a included yeast extract 14.44 g/L, molasses 19.14 g/L, and ammonium sulfate 0.20 g/L. In addition, the antifungal activity of P. polymyxa IN937a was also tested in this study. The preliminary results of in vitro antifungal activity indicated that P. polymyxa IN937a had a good inhibition on the growth of two phytopathogenic fungal strains Sclerotium rolfsii and Rhizoctonia solani. These results could be used for further research on the fermentation of P. polymyxa IN937a on a pilot scale to obtain the optimal number of spores for use in the development of biological crop protection products.
多粘类芽孢杆菌IN937a产孢子发酵培养基的优化及其抑菌活性
本研究对多粘类芽孢杆菌IN937a的发酵培养基进行了优化。通过Plackett-Burman实验,选取糖蜜、葡萄糖、硫酸镁、焦磷酸钾、酵母浸膏、硫酸锌、硫酸铵7个因子作为筛选P. polymyxa IN937a产孢影响因素的基础。Plackett-Burman基质分析结果表明,酵母浸膏、糖蜜和硫酸铵是影响P. polymyxa IN937a孢子产量的3个主要影响因子(P < 0.05)。然后,采用Box-Behnken设计,通过响应面法(RSM)优化3个因素的最佳组合,以提高多黏菌IN937a发酵的孢子产量。RSM法预测,在酵母膏14.44 g/L、糖蜜19.14 g/L、硫酸铵0.20 g/L的培养基条件下,实验48h后,多粘菌IN937a的最大孢子密度为6.606×109 spore/mL。此外,本研究还测试了P. polymyxa IN937a的抗真菌活性。体外抑菌活性初步结果表明,P. polymyxa IN937a对两种植物病原真菌菌核菌rolfsii和solani Rhizoctonia的生长有较好的抑制作用。这些结果可为进一步开展多粘菌IN937a的中试发酵研究,以获得最佳孢子数量,用于生物作物保护产品的开发。
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