Structural basis for the Ca2+-enhanced thermostability and activity of PET-degrading cutinase-like enzyme from Saccharomonospora viridis AHK190
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| 024 | 7 | 0 | |a 10.1007/s00253-014-6272-8 |2 doi |
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| 245 | 0 | 0 | |a Structural basis for the Ca2+-enhanced thermostability and activity of PET-degrading cutinase-like enzyme from Saccharomonospora viridis AHK190 |h [Elektronische Daten] |c [Takuya Miyakawa, Hiroki Mizushima, Jun Ohtsuka, Masayuki Oda, Fusako Kawai, Masaru Tanokura] |
| 520 | 3 | |a A cutinase-like enzyme from Saccharomonospora viridis AHK190, Cut190, hydrolyzes the inner block of polyethylene terephthalate (PET); this enzyme is a member of the lipase family, which contains an α/β hydrolase fold and a Ser-His-Asp catalytic triad. The thermostability and activity of Cut190 are enhanced by high concentrations of calcium ions, which is essential for the efficient enzymatic hydrolysis of amorphous PET. Although Ca2+-induced thermostabilization and activation of enzymes have been well explored in α-amylases, the mechanism for PET-degrading cutinase-like enzymes remains poorly understood. We focused on the mechanisms by which Ca2+ enhances these properties, and we determined the crystal structures of a Cut190 S226P mutant (Cut190S226P) in the Ca2+-bound and free states at 1.75 and 1.45Å resolution, respectively. Based on the crystallographic data, a Ca2+ ion was coordinated by four residues within loop regions (the Ca2+ site) and two water molecules in a tetragonal bipyramidal array. Furthermore, the binding of Ca2+ to Cut190S226P induced large conformational changes in three loops, which were accompanied by the formation of additional interactions. The binding of Ca2+ not only stabilized a region that is flexible in the Ca2+-free state but also modified the substrate-binding groove by stabilizing an open conformation that allows the substrate to bind easily. Thus, our study explains the structural basis of Ca2+-enhanced thermostability and activity in PET-degrading cutinase-like enzyme for the first time and found that the inactive state of Cut190S226P is activated by a conformational change in the active-site sealing residue, F106. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2014 | ||
| 690 | 7 | |a PET hydrolase |2 nationallicence | |
| 690 | 7 | |a Cutinase-type polyesterase |2 nationallicence | |
| 690 | 7 | |a Ca2+ activation |2 nationallicence | |
| 690 | 7 | |a Thermostability |2 nationallicence | |
| 690 | 7 | |a Saccharomonospora viridis AHK190 |2 nationallicence | |
| 700 | 1 | |a Miyakawa |D Takuya |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | |
| 700 | 1 | |a Mizushima |D Hiroki |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | |
| 700 | 1 | |a Ohtsuka |D Jun |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | |
| 700 | 1 | |a Oda |D Masayuki |u Laboratory of Biophysical Chemistry, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, 606-8522, Sakyo-ku, Kyoto, Japan |4 aut | |
| 700 | 1 | |a Kawai |D Fusako |u Center for Fiber and Textile Science, Kyoto Institute of Technology, Matsugasaki, 606-8585, Sakyo-ku, Kyoto, Japan |4 aut | |
| 700 | 1 | |a Tanokura |D Masaru |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/10(2015-05-01), 4297-4307 |x 0175-7598 |q 99:10<4297 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6272-8 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6272-8 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Miyakawa |D Takuya |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Mizushima |D Hiroki |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Ohtsuka |D Jun |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Oda |D Masayuki |u Laboratory of Biophysical Chemistry, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, 606-8522, Sakyo-ku, Kyoto, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Kawai |D Fusako |u Center for Fiber and Textile Science, Kyoto Institute of Technology, Matsugasaki, 606-8585, Sakyo-ku, Kyoto, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Tanokura |D Masaru |u Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, 113-8657, Bunkyo-ku, Tokyo, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/10(2015-05-01), 4297-4307 |x 0175-7598 |q 99:10<4297 |1 2015 |2 99 |o 253 | ||