caa a22 4500 397555598 CHVBK 20180308164801.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021529 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021529 Dipeptidyl Peptidase IV from Xanthomonas maltophilia: Sequencing and Expression of the Enzyme Gene and Characterization of the Expressed Enzyme [Elektronische Daten] [Tsutomu Kabashima, Kiyoshi Ito, Tadashi Yoshimoto] The dipeptidyl peptidase IV [EC 3.4.14.5] gene of Xanthomonas maltophilia, expressed in Escherichia coli, was cloned by the shotgun method. Nucleotide sequence analysis revealed an open reading frame of 2,223 bp, coding for a protein of 741 amino acids with a predicted molecular weight of 82,080. The expressed enzyme was extracted with SDS, and the solubilized enzyme was purified about 1,030-fold on columns of Toyopearl HW65C, DEAE Toyopearl twice, and hydroxyapatite, with an activity recovery of 50%. The enzyme hydrolyzed a proline-containing peptide at the penultimate position, and was inhibited by diisopropyl phosphofluoridate. The enzyme was most active at pH 8.5, and was stable between at pH 7.0 and 9.0. The molecular weight of the purified enzyme was estimated to be 83,000 and 165,000 by SDS-PAGE and gel filtration, respectively. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence α/βhydrolase fold family nationallicence cloning nationallicence crystallization nationallicence dipeptidyl peptidase IV nationallicence serine protease nationallicence Kabashima Tsutomu School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut Ito Kiyoshi School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut Yoshimoto Tadashi School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1111-1117 0021-924X 120:6<1111 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021529 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021529 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kabashima Tsutomu School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut NATIONALLICENCE 700 1- Ito Kiyoshi School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut NATIONALLICENCE 700 1- Yoshimoto Tadashi School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-macki, Nagasaki, Nagasaki 852 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1111-1117 0021-924X 120:6<1111 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford