Abstract
We cloned the β-glucuronidase gene (AtGUS) from Aspergillus terreus Li-20 encoding 657 amino acids (aa), which can transform glycyrrhizin into glycyrrhetinic acid monoglucuronide (GAMG) and glycyrrhetinic acid (GA). Based on sequence alignment, the C-terminal non-conservative sequence showed low identity with those of other species; thus, the partial sequence AtGUS(-3t) (1-592 aa) was amplified to determine the effects of the non-conservative sequence on the enzymatic properties. AtGUS and AtGUS(-3t) were expressed in E. coli BL21, producing AtGUS-E and AtGUS(-3t)-E, respectively. At the similar optimum temperature (55°C) and pH (AtGUS-E, 6.6; AtGUS(-3t)-E, 7.0) conditions, the thermal stability of AtGUS(-3t)-E was enhanced at 65°C, and the metal ions Co(2+), Ca(2+) and Ni(2+) showed opposite effects on AtGUS-E and AtGUS(-3t)-E, respectively. Furthermore, Km of AtGUS(-3t)-E (1.95 mM) was just nearly one-seventh that of AtGUS-E (12.9 mM), whereas the catalytic efficiency of AtGUS(-3t)-E was 3.2 fold higher than that of AtGUS-E (7.16 vs. 2.24 mM s(-1)), revealing that the truncation of non-conservative sequence can significantly improve the catalytic efficiency of AtGUS. Conformational analysis illustrated significant difference ...Continue Reading
References
May 1, 1992·Internal Medicine·A ChubachiA B Miura
Apr 1, 1991·Planta medica·H AkamatsuY Niwa
Oct 16, 1995·Gene·W P StemmerH L Heyneker
Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·B HenrissatG Davies
Jun 1, 1994·PCR Methods and Applications·R C Cadwell, G F Joyce
Jun 30, 1994·Nature·R H JacobsonB W Matthews
Apr 1, 1994·Planta medica·M NoseY Ogihara
Sep 15, 1995·Structure·G Davies, B Henrissat
Apr 1, 1996·Nature Structural Biology·S JainJ H Grubb
Apr 7, 1998·Nature Biotechnology·H ZhaoF H Arnold
Apr 5, 2000·The Journal of Biological Chemistry·M CicekA Esen
Jan 6, 2001·Biological & Pharmaceutical Bulletin·T Akao
Sep 12, 2001·Current Opinion in Biotechnology·B HöckerR Sterner
Feb 7, 2003·Current Medicinal Chemistry·L A Baltina
Jun 20, 2003·Lancet·J CinatlH W Doerr
Jun 11, 2004·The Journal of Antimicrobial Chemotherapy·Rea KrausseUwe Ullmann
Jan 13, 2005·Journal of Molecular Biology·Markus Wiederstein, Manfred J Sippl
Nov 10, 2005·Chemical Reviews·Reinhard Sterner, Birte Höcker
Jul 18, 2008·Bioresource Technology·Yujuan WangZengliang Yu
Feb 27, 2009·Molecular and Cellular Biochemistry·Xiu-Jie XieQun Wei
Feb 27, 2009·Cancer Research·Liat FuxNeta Ilan
Feb 2, 2010·Biochemical and Biophysical Research Communications·Frances H A HuismanEmily J Parker
Nov 6, 2010·Science·Bret D WallaceMatthew R Redinbo
Dec 1, 2006·Journal of Molecular Catalysis. B, Enzymatic·Ding-Qiang LuPing-Kai Ouyang
Citations
Jan 19, 2016·Enzyme and Microbial Technology·Nikou HesariMorteza Abbaszadegan
Dec 6, 2012·Bioresource Technology·Xiaoxiao GuoXiaohong Zhou
Aug 16, 2018·Applied Microbiology and Biotechnology·Bin WeiRu Yan
Jul 29, 2018·Applied and Environmental Microbiology·Yinghua XuChun Li
Jul 1, 2018·Comprehensive Reviews in Food Science and Food Safety·Lichun GuoWei Zhao