caa a22 4500 445326654 CHVBK 20180317142723.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s10989-011-9249-9 doi (NATIONALLICENCE)springer-10.1007/s10989-011-9249-9 Lactose Induction Increases Production of Recombinant Keratinocyte Growth Factor-2 in Escherichia coli [Elektronische Daten] [Haishan Tian, Lu Tang, Yi Wang, Xiaojie Wang, Lili Guan, Jian Zhang, Xiaoping Wu, Xiaokun Li] Recombinant human keratinocyte growth factor-2 (rhKGF-2) has previously been expressed in Escherichia coli using isopropyl-β-d-thiogalactopyranoside (IPTG), a non-metabolizable and expensive compound, as the inducer. In order to determine whether IPTG could be replaced with the cheap and natural lactose to induce rhKGF-2 expression, we examined the expression of rhKGF-2 in flask culture and 30-l fermentation using lactose as the inducer. The optimized fermentation induced with lactose resulted in 1,382g of cell mass, corresponding to a 84% enhancement in cell mass compared with IPTG induction. While the expression level of rhKGF-2 induced with lactose was comparable to that induced with IPTG, the solubility of target protein was increased by lactose induction than by IPTG induction. The recombinant protein was further purified by cation exchange and heparin-affinity chromatography. 255 milligrams of pure rhKGF-2 was achieved per liter culture by lactose induction, 52% higher than that obtained by IPTG induction. A preliminary biochemical characterization of purified rhKGF-2 was performed by Western blotting and mitogenic activity analysis, and the results demonstrated that the purified lactose-induced rhKGF-2 could react with anti-human KGF-2 antibody and stimulate the proliferation of FGFR2-IIIb-transfected mouse BaF3 cells as IPTG-induced rhKGF-2 could do. Springer Science+Business Media, LLC, 2011 Human keratinocyte growth factor-2 nationallicence Lactose induction nationallicence Fed-batch nationallicence Solubility nationallicence Mitogenic activity nationallicence Tian Haishan Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut Tang Lu School of Pharmaceutical Science, Wenzhou Medical College, 325035, Wenzhou, Zhejiang, China aut Wang Yi Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut Wang Xiaojie School of Pharmaceutical Science, Wenzhou Medical College, 325035, Wenzhou, Zhejiang, China aut Guan Lili Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut Zhang Jian Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut Wu Xiaoping Institute of Tissue Transplantation and Immunology, Jinan University, 510632, Guangzhou, Guangdong, China aut Li Xiaokun Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut International Journal of Peptide Research and Therapeutics Springer Netherlands 17/2(2011-06-01), 123-129 1573-3149 17:2<123 2011 17 10989 https://doi.org/10.1007/s10989-011-9249-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10989-011-9249-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tian Haishan Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut NATIONALLICENCE 700 1- Tang Lu School of Pharmaceutical Science, Wenzhou Medical College, 325035, Wenzhou, Zhejiang, China aut NATIONALLICENCE 700 1- Wang Yi Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut NATIONALLICENCE 700 1- Wang Xiaojie School of Pharmaceutical Science, Wenzhou Medical College, 325035, Wenzhou, Zhejiang, China aut NATIONALLICENCE 700 1- Guan Lili Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut NATIONALLICENCE 700 1- Zhang Jian Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut NATIONALLICENCE 700 1- Wu Xiaoping Institute of Tissue Transplantation and Immunology, Jinan University, 510632, Guangzhou, Guangdong, China aut NATIONALLICENCE 700 1- Li Xiaokun Engineering Research Center of Bioreactor and Pharmaceutical Development of Ministry of Education, School of Life Sciences, Jilin Agricultural University, 130118, Changchun, Jilin, China aut NATIONALLICENCE 773 0- International Journal of Peptide Research and Therapeutics Springer Netherlands 17/2(2011-06-01), 123-129 1573-3149 17:2<123 2011 17 10989 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer