MaghrebDataLibMedV2, Main, Exploration, bibRecord, 000208

Thermostability improvement of maltogenic amylase MAUS149 by error prone PCR.

Identifieur interne : 000208 ( Main/Exploration ); précédent : 000207; suivant : 000209

Thermostability improvement of maltogenic amylase MAUS149 by error prone PCR.

Auteurs : Sameh Ben Mabrouk [Tunisie] ; Dorra Zouari Ayadi ; Hajer Ben Hlima ; Samir Bejar

Source :

Journal of biotechnology [ 1873-4863 ] ; 2013.

RBID : pubmed:23994264

Descripteurs français

KwdFr :
- Bacillus (enzymologie), Glycosidases (composition chimique), Glycosidases (génétique), Mutagenèse dirigée (MeSH), Mutation (MeSH), Réaction de polymérisation en chaîne (méthodes), Stabilité enzymatique (MeSH), Séquence d'acides aminés (génétique), Température élevée (MeSH).
MESH :
- composition chimique : Glycosidases.
- enzymologie : Bacillus.
- génétique : Glycosidases, Séquence d'acides aminés.
- méthodes : Réaction de polymérisation en chaîne.
- Mutagenèse dirigée, Mutation, Stabilité enzymatique, Température élevée.

English descriptors

KwdEn :
- Amino Acid Sequence (genetics), Bacillus (enzymology), Enzyme Stability (MeSH), Glycoside Hydrolases (chemistry), Glycoside Hydrolases (genetics), Hot Temperature (MeSH), Mutagenesis, Site-Directed (MeSH), Mutation (MeSH), Polymerase Chain Reaction (methods).
MESH :
- chemical , chemistry : Glycoside Hydrolases.
- enzymology : Bacillus.
- genetics : Amino Acid Sequence, Glycoside Hydrolases.
- methods : Polymerase Chain Reaction.
- Enzyme Stability, Hot Temperature, Mutagenesis, Site-Directed, Mutation.

Abstract

The thermostability of maltogenic amylase from Bacillus sp. US149 (MAUS149) was improved by random mutagenesis using error prone PCR. The library constructed for the mutants obtained was subjected to screening, leading to the selection of a thermostable mutant enzyme named MA-A27. The latter was noted to contain four single mutations, namely D46V, P78L, V145A, and K548E. The half-life times recorded for MA-A27 at 50°C and 55°C were 70 min and 25 min, compared to 30 min and 13 min for the wild type, respectively. The results from molecular modeling attributed the increase in thermostability observed for MA-A27 to P78L and K548E substitutions that led to new hydrogen bond and salt bridge formations. Further site-directed mutagenesis studies showed that the P78L and K548E single mutations underwent an increase in thermostability, thus confirming the joint contribution of both substitutions to the increase in thermostability observed for MA-A27.

DOI: 10.1016/j.jbiotec.2013.08.026
PubMed: 23994264

Affiliations:

Tunisie

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Le document en format XML

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<front><div type="abstract" xml:lang="en">The thermostability of maltogenic amylase from Bacillus sp. US149 (MAUS149) was improved by random mutagenesis using error prone PCR. The library constructed for the mutants obtained was subjected to screening, leading to the selection of a thermostable mutant enzyme named MA-A27. The latter was noted to contain four single mutations, namely D46V, P78L, V145A, and K548E. The half-life times recorded for MA-A27 at 50°C and 55°C were 70 min and 25 min, compared to 30 min and 13 min for the wild type, respectively. The results from molecular modeling attributed the increase in thermostability observed for MA-A27 to P78L and K548E substitutions that led to new hydrogen bond and salt bridge formations. Further site-directed mutagenesis studies showed that the P78L and K548E single mutations underwent an increase in thermostability, thus confirming the joint contribution of both substitutions to the increase in thermostability observed for MA-A27.</div>
</front>
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<name sortKey="Ben Hlima, Hajer" sort="Ben Hlima, Hajer" uniqKey="Ben Hlima H" first="Hajer" last="Ben Hlima">Hajer Ben Hlima</name>
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Thermostability improvement of maltogenic amylase MAUS149 by error prone PCR.

Thermostability improvement of maltogenic amylase MAUS149 by error prone PCR.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki