CSIR - Institute of Microbial Technology, Sector 39-A, Chandigarh 160036, India.
Bioresour Technol. 2012 Nov;123:135-43. doi: 10.1016/j.biortech.2012.07.011. Epub 2012 Jul 16.
In this study, effects of temperature, inducer concentration, time of induction and co-expression of molecular chaperones (GroEL-GroES and DnaKJE), on cell growth and solubilization of model protein, xylanases, were investigated. The yield of soluble xylanases increased with decreasing cultivation temperature and inducer level. In addition, co-expression of DnaKJE chaperone resulted in increased soluble xylanases though the time of induction of chaperone and target protein had a bearing on this yield. A combination of chaperone co-expression and partial induction resulted in ∼40% (in DnaKJE) and 33% (in GroEL-GroES) of total xylanase yield in soluble fraction. However, the conditions for maximum yield of soluble r-XynB and maximum % soluble expression of r-XynB were different. Higher expression of soluble xylanases in a scalable semi-synthetic medium showed potential of the process for soluble enzyme production.
在这项研究中，考察了温度、诱导剂浓度、诱导时间以及分子伴侣（GroEL-GroES 和 DnaKJE）共表达对细胞生长和模型蛋白木聚糖酶可溶性的影响。随着培养温度和诱导剂水平的降低，可溶性木聚糖酶的产率增加。此外，尽管诱导伴侣蛋白和目标蛋白的时间会影响产量，但共表达 DnaKJE 伴侣蛋白会导致可溶性木聚糖酶产量增加。伴侣蛋白共表达和部分诱导的组合使可溶性部分的总木聚糖酶产量达到约 40%（在 DnaKJE 中）和 33%（在 GroEL-GroES 中）。然而，可溶性 r-XynB 最大产量和 r-XynB 最大可溶性表达的条件不同。在可扩展的半合成培养基中表达更高水平的可溶性木聚糖酶表明该工艺具有可溶性酶生产的潜力。
