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Abstract <p>Food crops are the indispensable part of human diet, however, crop plants are continuously at the risk of attack by pathogenic fungi. To counter such attack, plants produce pathogenesis related (PR) proteins and phytoalexins. Among these, β-1,3-glucanases play an indispensable role by hydrolysing β-glucans in fungal cell wall to enhance the plant’s resistance against fungal attack. Hence, the present study aims to report the cloning, sub-cloning, recombinant expression and <i>in silico</i> characterization of β-1,3-glucanase (<i>TaGlb2a</i>) gene from wheat. Purified recombinant β-1,3-glucanase possessed 34.6 U/mg specific activity with 45% recovery and displayed a ~38 kDa band on SDS-PAGE and zymogram analysis. The <i>in silico</i> studies predicted TaGlb2a protein sequence to be 342 amino acids long, containing a conserved glycosyl hydrolase family 17 (GH17) domain. The theoretical isoelectric point (pI) and molecular weight were computed to be 4.67 and 31 788 Da, respectively. Secondary structure analysis indicated the predominance of random coil sequences, followed by α-helixes and extended chains. The BLASTp analysis of TaGlb2a confirmed greater sequence similarity with <i>Hordeum vulgare</i> (84.84%), <i>Sorghum bicolor</i> (64.80%), <i>Avena sativa</i> (57.7%). The phylogenetic analysis clustered, all the sequences in single clade, displaying high structural and functional similarity against biotic stresses. Further studies would unlock valuable information regarding efficient utilization of recombinant β-1,3-glucanases in the improvement of fungal resistance in different plants varieties. Additionally, this enzyme may possess considerable potential in brewing industry due to its role in hydrolyzing grain cell wall β-glucans.</p>

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Heterologous Expression of Triticum aestivum β-1,3-Glucanase Taglb2a Gene in E. coli and in silico Structural Characterization

  • 期刊论文

  • 发表日期：

    2025年12月1日

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  • Fatima Shahid,
  • Hamama Islam Butt,
  • Hira Muzzamal,
  • Beenish Maqsood,
  • Asima Ayyub,
  • Riffat Tahira,
  • Mahjabeen Saleem

摘要

Abstract

Food crops are the indispensable part of human diet, however, crop plants are continuously at the risk of attack by pathogenic fungi. To counter such attack, plants produce pathogenesis related (PR) proteins and phytoalexins. Among these, β-1,3-glucanases play an indispensable role by hydrolysing β-glucans in fungal cell wall to enhance the plant’s resistance against fungal attack. Hence, the present study aims to report the cloning, sub-cloning, recombinant expression and in silico characterization of β-1,3-glucanase (TaGlb2a) gene from wheat. Purified recombinant β-1,3-glucanase possessed 34.6 U/mg specific activity with 45% recovery and displayed a ~38 kDa band on SDS-PAGE and zymogram analysis. The in silico studies predicted TaGlb2a protein sequence to be 342 amino acids long, containing a conserved glycosyl hydrolase family 17 (GH17) domain. The theoretical isoelectric point (pI) and molecular weight were computed to be 4.67 and 31 788 Da, respectively. Secondary structure analysis indicated the predominance of random coil sequences, followed by α-helixes and extended chains. The BLASTp analysis of TaGlb2a confirmed greater sequence similarity with Hordeum vulgare (84.84%), Sorghum bicolor (64.80%), Avena sativa (57.7%). The phylogenetic analysis clustered, all the sequences in single clade, displaying high structural and functional similarity against biotic stresses. Further studies would unlock valuable information regarding efficient utilization of recombinant β-1,3-glucanases in the improvement of fungal resistance in different plants varieties. Additionally, this enzyme may possess considerable potential in brewing industry due to its role in hydrolyzing grain cell wall β-glucans.