Bioengineering Laboratory, Department of Chemical Engineering, Monash University, Wellington Road, Clayton, VIC 3800, Australia.
Appl Biochem Biotechnol. 2012 Nov;168(5):1035-50. doi: 10.1007/s12010-012-9839-9. Epub 2012 Aug 16.
Using a combination of conventional sequential techniques, the batch growth conditions for the production of cell-envelope-associated proteinases have for the first time been studied and optimised in Lactobacillus delbrueckii subsp. lactis 313 (ATCC 7830; LDL 313). Concentrations of inoculum (0.1 < X < 10 % vol/vol), agitation speed (0 < S < 200 rpm), varying incubation temperature (30 < T < 50 °C), starting pH (4.5 < pH < 7) and carbon/nitrogen ratio of production medium (0.2 < r < 5) had an individual effect on proteinase yield (p < 0.01). Optimal conditions for proteinase production included an initial pH of 6.0, 45 °C incubation temperature, 2 % (v/v) inoculum size of OD(560) = 1, 150 rpm agitation speed, and growth medium carbon/nitrogen ratio of 1.0. Maximum proteinase activity obtained for whole cells was 0.99 U/ml after 8 h of incubation. The variables studied are very relevant due to their significance in improving the productivity of proteinase synthesis from LDL 313, under process and, likely, economic optimum conditions.
首次研究并优化了德氏乳杆菌亚种。乳脂亚种 313(ATCC 7830;LDL 313)生产细胞包膜相关蛋白酶的分批生长条件,采用常规顺序技术组合。接种物浓度(0.1 < X < 10 vol/vol)、搅拌速度(0 < S < 200 rpm)、不同的培养温度(30 < T < 50°C)、起始 pH 值(4.5 < pH < 7)和生产培养基的碳/氮比(0.2 < r < 5)对蛋白酶产量均有单独影响(p < 0.01)。蛋白酶生产的最佳条件包括初始 pH 值为 6.0、45°C 培养温度、2%(v/v)OD(560)= 1 的接种量、150 rpm 的搅拌速度和生长培养基的碳/氮比为 1.0。培养 8 小时后,整个细胞获得的最大蛋白酶活性为 0.99 U/ml。研究的变量非常重要,因为它们对提高 LDL 313 中蛋白酶合成的生产力具有重要意义,在工艺和经济最佳条件下也是如此。