Microbial Sciences Division, Agharkar Research Institute, Pune, India.
J Basic Microbiol. 2012 Oct;52(5):539-48. doi: 10.1002/jobm.201100446. Epub 2012 Feb 23.
The influence of concentration of medium components such as colloidal chitin, lactose, malt extract, yeast extract, and peptone on the chitinase production from Basidiobolous ranarum at the flask level were studied by using statistical tool Central Composite Design (CCD) and analysed by Response Surface Methodology (RSM). The results revealed that colloidal chitin, malt extract and peptone had significant effect (P < 0.01) on the chitinase production at their individual levels. The polynomial equation of the model developed incorporates 3 linear, 3 quadratic and 5 interactive terms. Maximum chitinase production of 3.47 U ml(-1) was achieved with 1.5% colloidal chitin, 0.125% lactose, 0.025% malt extract and 0.075% peptone. After optimization, chitinase activity was increased by 7.71 fold. A second order polynomial equation was found to be useful for the development of efficient bioprocess for chitinase production. To screen the biotechnological potential of this enzyme, degradation of fungal mycelia by ammonium sulphate precipitate of the same was studied for several pathogenic fungi-in vitro which showed promising results particularly against Rhizoctonia solani and Fusarium solani. This study provides the first evidence showing the effectiveness of RSM for the development of a robust statistical model for the chitinase production by Basidiobolus and for its application in the biocontrol of phytopathogenic fungi.
采用统计工具中心复合设计(CCD)研究了介质成分(胶体几丁质、乳糖、麦芽提取物、酵母提取物和蛋白胨)浓度对Ranarum basidiobolus 产几丁质酶的影响,并通过响应面法(RSM)进行了分析。结果表明,胶体几丁质、麦芽提取物和蛋白胨在各自的水平上对几丁质酶的产生有显著影响(P < 0.01)。所建立的模型多项式方程包含 3 个线性项、3 个二次项和 5 个交互项。在 1.5%胶体几丁质、0.125%乳糖、0.025%麦芽提取物和 0.075%蛋白胨的条件下,几丁质酶的最大产量为 3.47 U ml(-1)。优化后,几丁质酶活性提高了 7.71 倍。发现二次多项式方程可用于开发有效的几丁质酶生产生物工艺。为了筛选该酶的生物技术潜力,研究了相同的硫酸铵沉淀对几种病原真菌的菌丝体的降解-体外结果表明,该酶对 Rhizoctonia solani 和 Fusarium solani 具有特别有希望的效果。本研究首次证明了 RSM 用于开发 Ranarum basidiobolus 几丁质酶生产的稳健统计模型及其在植物病原真菌生物防治中的应用的有效性。