Pozdnyakov Nikita, Shilov Sergey, Lukin Alexander, Bolshakov Maxim, Sogorin Evgeny
Federal Research Center "Pushchino Scientific Center for Biological Research of the RAS", Institute for Biological Instrumentation, Institutskaya Street, 142290, Pushchino, Russia.
Federal Research Center "Pushchino Scientific Center for Biological Research of the RAS", Institute of Basic Biological Problems, Institutskaya Street, 142290, Pushchino, Russia.
Bioresour Bioprocess. 2022 Apr 4;9(1):37. doi: 10.1186/s40643-022-00518-2.
Using parameters of optimal conditions from laboratory experiments often results in the loss of significant time and resources when trying to scale up the process. In this study, the comparison of results of laboratory and semi-industrial experiments of enzymatic hydrolysis of soy protein isolate is considered. The kinetics of peptides accumulation was investigated by colorimetric method in both microtube (volume reaction is 0.7 ml, 7.14 mg/ml of substrate, incubation in solid state thermostat) and industrial homogenizer (volume reaction is 4,000 ml, 100 mg/ml of substrate, rotor-stator type mixer). The enzyme preparation Protosubtilin G3x (main component is subtilisin) was used as an analogue of the Alcalase preparation, which is already widely used in the food industry. It was found that the pH and the number of proteolytic units in the reaction mixture of both scales had slightly different results of the kinetics, while the temperature showed significantly one. The laboratory scale of the reaction had a wide range of optimal temperature (40-60 C, 30 C showed slowest rate of kinetics reaction), whereas the semi-industrial scale had 50 C of optimal temperature (30, 40, 60 C had the same kinetics). It also was found that maintaining the pH value of the reaction mixture was not mandatory. The obtained results indicate the need to refine the process conditions using semi-industrial experiments before attracting industrial-scale resources. In the case of selection of conditions for the hydrolysis of soy protein isolate in production, it is necessary first of all to take into account the reaction temperature as the most irreproducible parameter when scaling.
在试图扩大工艺规模时，使用实验室实验中的最佳条件参数往往会导致大量时间和资源的浪费。本研究考虑了大豆分离蛋白酶解的实验室和半工业实验结果的比较。通过比色法在微量管（反应体积为0.7毫升，底物浓度为7.14毫克/毫升，在固态恒温器中孵育）和工业均质器（反应体积为4000毫升，底物浓度为100毫克/毫升，转子-定子型混合器）中研究了肽积累的动力学。酶制剂Protosubtilin G3x（主要成分是枯草杆菌蛋白酶）被用作Alcalase制剂的类似物，Alcalase制剂已在食品工业中广泛使用。结果发现，两种规模反应混合物中的pH值和蛋白水解单元数量在动力学结果上略有不同，而温度则显示出显著差异。实验室规模的反应具有较宽的最佳温度范围（40-60℃，30℃时动力学反应速率最慢），而半工业规模的最佳温度为50℃（30、40、60℃时动力学相同）。还发现保持反应混合物的pH值并非必要。所得结果表明，在引入工业规模资源之前，需要通过半工业实验来优化工艺条件。在选择大豆分离蛋白生产中的水解条件时，首先必须考虑反应温度，因为它是扩大规模时最不可重复的参数。
