Park Jin-Seung, Han Kyung-Yeon, Song Jong-Am, Ahn Keum-Young, Seo Hyuk-Seong, Lee Jeewon
Department of Chemical and Biological Engineering, Korea University, Anam-Dong 5-1, Sungbuk-Ku, Seoul, 136-713, South Korea.
Biotechnol Lett. 2007 Oct;29(10):1513-8. doi: 10.1007/s10529-007-9417-3. Epub 2007 Jun 5.
Using 2-dimensional gel electrophoresis, the Escherichia coli proteome response to a heat-shock stress was analyzed and a 1.6-fold increase of malate dehydrogenase was observed even under the heat-shock condition where the total number of soluble proteins decreased by about 5%. We subsequently demonstrated that, as an N-terminus fusion expression partner, malate dehydrogenase facilitated the folding of, and dramatically increased the solubility of, many aggregation-prone heterologous proteins in E. coli cytoplasm. Therefore, malate dehydrogenase is well suited for production of a biologically active fusion mutant of cutinase (Pseudomonas putida origin) that is currently of considerable to biotechnology and commercial industries.
利用二维凝胶电泳分析了大肠杆菌蛋白质组对热休克应激的反应，结果发现，即使在热休克条件下可溶性蛋白质总数减少了约5%，苹果酸脱氢酶的含量仍增加了1.6倍。我们随后证明，作为N端融合表达伙伴，苹果酸脱氢酶促进了许多易于聚集的异源蛋白在大肠杆菌细胞质中的折叠，并显著提高了其溶解度。因此，苹果酸脱氢酶非常适合用于生产角质酶（源自恶臭假单胞菌）的生物活性融合突变体，该突变体目前对生物技术和商业行业具有重要意义。
