Patil Mahesh D, Patel Gopal, Surywanshi Balaji, Shaikh Naeem, Garg Prabha, Chisti Yusuf, Banerjee Uttam Chand
Department of Pharmaceutical Technology (Biotechnology), National Institute of Pharmaceutical Education and Research, Sector-67, S.A.S. Nagar, Punjab, 160062, India.
Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research, Sector-67, S.A.S. Nagar, Punjab, 160062, India.
AMB Express. 2016 Dec;6(1):84. doi: 10.1186/s13568-016-0260-6. Epub 2016 Oct 3.
Disruption of Pseudomonas putida KT2440 by high-pressure homogenization in a French press is discussed for the release of arginine deiminase (ADI). The enzyme release response of the disruption process was modelled for the experimental factors of biomass concentration in the broth being disrupted, the homogenization pressure and the number of passes of the cell slurry through the homogenizer. For the same data, the response surface method (RSM), the artificial neural network (ANN) and the support vector machine (SVM) models were compared for their ability to predict the performance parameters of the cell disruption. The ANN model proved to be best for predicting the ADI release. The fractional disruption of the cells was best modelled by the RSM. The fraction of the cells disrupted depended mainly on the operating pressure of the homogenizer. The concentration of the biomass in the slurry was the most influential factor in determining the total protein release. Nearly 27 U/mL of ADI was released within a single pass from slurry with a biomass concentration of 260 g/L at an operating pressure of 510 bar. Using a biomass concentration of 100 g/L, the ADI release by French press was 2.7-fold greater than in a conventional high-speed bead mill. In the French press, the total protein release was 5.8-fold more than in the bead mill. The statistical analysis of the completely unseen data exhibited ANN and SVM modelling as proficient alternatives to RSM for the prediction and generalization of the cell disruption process in French press.
讨论了在法国压榨机中通过高压均质化破坏恶臭假单胞菌KT2440以释放精氨酸脱亚氨酶(ADI)的情况。针对被破坏肉汤中的生物质浓度、均质化压力以及细胞浆液通过均质器的次数等实验因素,对破坏过程中的酶释放响应进行了建模。对于相同的数据,比较了响应面法(RSM)、人工神经网络(ANN)和支持向量机(SVM)模型预测细胞破坏性能参数的能力。结果证明ANN模型最适合预测ADI释放。细胞的部分破坏情况用RSM建模效果最佳。细胞被破坏的比例主要取决于均质器的操作压力。浆液中生物质的浓度是决定总蛋白释放的最有影响的因素。在操作压力为510巴、生物质浓度为260 g/L的浆液中单次通过后可释放出近27 U/mL的ADI。使用100 g/L的生物质浓度时,法国压榨机释放的ADI比传统高速珠磨机高2.7倍。在法国压榨机中,总蛋白释放量比珠磨机多5.8倍。对完全未见过的数据进行的统计分析表明,ANN和SVM建模是RSM的有效替代方法,可用于预测和推广法国压榨机中的细胞破坏过程。