Bioinformatics Analysis of Upstream Region and Protein Structure of Fungal Phytase Gene

Document Type : Short Communication

Authors

1 Department of Animal Sciences, Faculty of Animal and Food Sciences, Khuzestan Ramin Agriculture and Natural Resources University, Mollasani, Ahvaz, Iran

2 Department of Genetics, Faculty of Animal Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

3 Department of Medical Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Cell and Molecular Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Phytase increases the bioavailability of phytate phosphorus in seed-based animal feeds and reduces the phosphorus pollution of animal waste. Since most animal feeds for pellets are heated up to 65-80 °C, the production of a thermostable structure for phytase can be useful. In this study, we sought to perform bioinformatics analysis of the upstream region and protein structure of fungal phytase to improve its expression and thermostability properties. We used bioinformatics methods such as similarity search, multiple alignment, statistical analysis of physicochemical properties of amino acids, pattern recognition, and protein modeling to find out the effective factors in heat resistance of phytase. Change in Gibbs free energy (ΔG) of the best pattern promoter resulting from the interaction between RNA polymerase and the promoter sequences of modified genes of phytase was equal to -9 kcalmol-1, which is lower compared to other interactions. The evaluation of the three-dimensional structure of new phytases showed that amino acid substitutions aimed at improving thermostability did not change the form and structure of the protein. The results of Prochek, Whatcheck, and ERRAT for structural analysis and verification were 84, 72, and 70, respectively, that were satisfactory.

Keywords


Article Title [French]

Analyse Bio-informatique de la Région en Amont du Gène de Phytase Fongique Ainsi que de sa Structure Protéique

Abstract [French]

La phytase augmente la biodisponibilité du phytate phosphore dans les graines dédiées à l’alimentation animale et réduit la pollution au phosphore des déchets animaliers. Étant donné que la plupart des aliments pour animaux destinés à la production de granulés sont chauffés à 65-80 °C, la production d'une structure thermostable pour la phytase peut être utile. Notre étude avait pour objectif l’analyse bioinformatique de la région en amont du gène de la phytase fongique ainsi que la sturucture de la protéine codée, afin d'améliorer ses propriétés d'expression et de thermostabilité. A cet effet, des méthodes bio-informatiques telles que la recherche de similarité, l'alignement multiple, l'analyse statistique des propriétés physicochimiques des acides aminés, la reconnaissance des formes et la modélisation des protéines pour déterminer les facteurs de résistance à la chaleur de la phytase ont été utilisées. La modification (ΔG) dans l'énergie libre de Gibbs du meilleur promoteur de modèle résultant de l'interaction entre l'ARN polymérase et les séquences promotrices de gènes modifiés de la phytase était égal à -9 kcalmol-1, ce qui est inférieur aux autres connexions. L’évaluation de la structure tridimensionnelle de nouvelles phytases a montré que les substitutions d’acides aminés visant à améliorer la thermostabilité n’ont pas modifié la forme et la structure de la protéine. Les résultats obtenus par Prochek, Whatcheck et ERRAT pour l’analyse structurelle étaient satisfaisants et la vérification était respectivement de 84, 72 et 70.

Keywords [French]

  • Phytase Fongique
  • Bio-Informatique
  • Modélisation Homologique
  • Amarrage Moléculaire
  • Analyse des Régions en Amont
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