Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.

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 ide...

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Autores principales:	Yanli Liu, Jie Huangfu, Feng Qi, Imdad Kaleem, Wenwen E, Chun Li
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d61e85a220e04ed6905e0d61de906a41
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spelling	oai:doaj.org-article:d61e85a220e04ed6905e0d61de906a412021-11-18T07:28:46ZEffects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.1932-620310.1371/journal.pone.0030998https://doaj.org/article/d61e85a220e04ed6905e0d61de906a412012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22347419/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203We 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 in the secondary structure between AtGUS-E and AtGUS(-3t)-E by circular dichroism (CD). The results showed that the truncation of the non-conservative sequence could preferably alter and influence the stability and catalytic efficiency of enzyme.Yanli LiuJie HuangfuFeng QiImdad KaleemWenwen EChun LiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 2, p e30998 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yanli Liu Jie Huangfu Feng Qi Imdad Kaleem Wenwen E Chun Li Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
description	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 in the secondary structure between AtGUS-E and AtGUS(-3t)-E by circular dichroism (CD). The results showed that the truncation of the non-conservative sequence could preferably alter and influence the stability and catalytic efficiency of enzyme.
format	article
author	Yanli Liu Jie Huangfu Feng Qi Imdad Kaleem Wenwen E Chun Li
author_facet	Yanli Liu Jie Huangfu Feng Qi Imdad Kaleem Wenwen E Chun Li
author_sort	Yanli Liu
title	Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
title_short	Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
title_full	Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
title_fullStr	Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
title_full_unstemmed	Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.
title_sort	effects of a non-conservative sequence on the properties of β-glucuronidase from aspergillus terreus li-20.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/d61e85a220e04ed6905e0d61de906a41
work_keys_str_mv	AT yanliliu effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20 AT jiehuangfu effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20 AT fengqi effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20 AT imdadkaleem effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20 AT wenwene effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20 AT chunli effectsofanonconservativesequenceonthepropertiesofbglucuronidasefromaspergillusterreusli20
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Effects of a non-conservative sequence on the properties of β-glucuronidase from Aspergillus terreus Li-20.

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