Cloning and analysis of pyrG gene encoding orotidine 5-monophosphate decarboxylase of Aspergillus oryzae strain Sl

Selina Oh Siew Ling, Leong Jiun Min, Abdul Munir Abd. Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In this study, the pyrG gene which encodes for orotidine 5-monophosphate decarboxylase (OMP decarboxylase) of Aspergillus oryzae strain Sl was cloned and analysed. This 1.8kb A. oryzae pyrG encompasses the 5'-regulatory flanking region (465 bp), open reading frame (899 bp) and 3'-regulatory region (475 bp). The pyrG contained one intron at position 623-687 bp based on the AUGUSTUS and FGENESH (SoftBerry) analysis corresponding to the intron present in the pyrG of A. oryzae (Accession Number: Y13811 ). In silico analysis showed that the enzyme encoded by the A. oryzae Sl pyrG gene has a theoretical molecular weight of 30.28 kDa and theoretical pl value of 5.92. This enzyme is hydrophilic, located in a region outside of the transmembrane and it functions in the cytoplasm. Five motives such as N-glycosylation sitef protein kinase C (PKC) phosphorylation site, casein kinase II (CK-2) phosphorylation site, N-myristolation site and orotidine 5-monophoshate decarboxylase active site have been identified in the pyrG amino acid sequence. The three dimensional structure of this enzyme generated via protein homology modeling using the bioinformatic software, Swiss Model, shows that OMP decarboxylase is a protein with an alpha;/beta; barrel structure possessing 8β-strands surrounded by 9 α-helices. The amino acid residues involved in the active site have been identified and it is located on one of the β-strands. The pyrG DNA sequence will be used for the complementation of a pyrG auxotroph mutant of A. oryzae.

Original languageEnglish
Pages (from-to)331-337
Number of pages7
JournalSains Malaysiana
Volume40
Issue number4
Publication statusPublished - Apr 2011

Fingerprint

Aspergillus oryzae
molecular cloning
genes
active sites
introns
phosphorylation
enzymes
auxotrophs
protein kinase C
glycosylation
bioinformatics
open reading frames
amino acid sequences
cytoplasm
proteins
orotidine-5'-phosphate decarboxylase
molecular weight
nucleotide sequences
mutants
amino acids

Keywords

  • Aspergillus oryzae
  • Orotidine 5-monophosphate dehydrogenase
  • pyrG

ASJC Scopus subject areas

  • General

Cite this

Cloning and analysis of pyrG gene encoding orotidine 5-monophosphate decarboxylase of Aspergillus oryzae strain Sl. / Ling, Selina Oh Siew; Min, Leong Jiun; Abd. Murad, Abdul Munir; Mahadi, Nor Muhammad; Abu Bakar, Farah Diba.

In: Sains Malaysiana, Vol. 40, No. 4, 04.2011, p. 331-337.

Research output: Contribution to journalArticle

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abstract = "In this study, the pyrG gene which encodes for orotidine 5-monophosphate decarboxylase (OMP decarboxylase) of Aspergillus oryzae strain Sl was cloned and analysed. This 1.8kb A. oryzae pyrG encompasses the 5'-regulatory flanking region (465 bp), open reading frame (899 bp) and 3'-regulatory region (475 bp). The pyrG contained one intron at position 623-687 bp based on the AUGUSTUS and FGENESH (SoftBerry) analysis corresponding to the intron present in the pyrG of A. oryzae (Accession Number: Y13811 ). In silico analysis showed that the enzyme encoded by the A. oryzae Sl pyrG gene has a theoretical molecular weight of 30.28 kDa and theoretical pl value of 5.92. This enzyme is hydrophilic, located in a region outside of the transmembrane and it functions in the cytoplasm. Five motives such as N-glycosylation sitef protein kinase C (PKC) phosphorylation site, casein kinase II (CK-2) phosphorylation site, N-myristolation site and orotidine 5-monophoshate decarboxylase active site have been identified in the pyrG amino acid sequence. The three dimensional structure of this enzyme generated via protein homology modeling using the bioinformatic software, Swiss Model, shows that OMP decarboxylase is a protein with an alpha;/beta; barrel structure possessing 8β-strands surrounded by 9 α-helices. The amino acid residues involved in the active site have been identified and it is located on one of the β-strands. The pyrG DNA sequence will be used for the complementation of a pyrG auxotroph mutant of A. oryzae.",
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AU - Mahadi, Nor Muhammad

AU - Abu Bakar, Farah Diba

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