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<p>Glucan-type exopolysaccharides (EPSs) produced by lactic acid bacteria (LAB) have gained considerable attention owing to the increasing demand for natural resources in food and pharmaceutical industries. LAB use glucosyltransferases (GTFs) to synthesize glucan-type EPSs with different sizes and structures. Exploring new EPS-producing sources is key to discovering genes responsible for diverse EPS types. Recently, the complete genomes of two novel LAB species, <i>Apilactobacillus waqarii</i> <b>HBW1</b> and <i>Apilactobacillus iqraium</i> <b>F1</b> were reported. This study aims to identify and characterize EPS producing GTF genes in these species using computational approaches. RAST annotation revealed the presence of three EPS synthesizing genes each in <i>A. waqarii</i>&#xa0;(designated as AwGTF1, AwGTF2 and AwGTF3) and <i>A. iqraium,</i> (designated as GTFG1-F1, GTFG2-F1 and GTFG3-F1). Evolutionary analysis indicated that most of the retrieved GTFs shared low similarity with known GTFs and formed a distinct clade closely adjacent to dextran-producing&#xa0;<i>L. mesenteroides</i>, along with <i>A. kunkeei</i>. Domain analysis revealed a conserved domain architecture, though AwGTF3 lacks a glucan-binding domain (GBD). Due to the high molecular weight of GH70 family GTFs, 3D structure prediction was only possible using recently discovered computational tools, which revealed structural variations among these GTFs. Our findings suggest that out of six identified GTFs in <i>A. waqarii</i>&#xa0;and <i>A. iqraium,</i> five are putative glucansucrases while AwGTF3, contain a tryptophan residue in place of glycine, may function as a branching sucrase. Further studies involving recombinant gene expression and functional characterization could reveal detailed structure–function relationship of these enzymes.</p>

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Deciphering the structural–functional dynamics of glucosyltransferases from novel Apilactobacillus isolates through in silico analysis

  • 期刊论文

  • 发表日期：

    2026年7月29日

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  • Hira Kiran,
  • Munir A. Anwar,
  • Waqar Ahmad,
  • Iqra Jawad,
  • Ayesha Siddiqa,
  • Muhammad Imran,
  • Moazur Rahman,
  • Muhammad Saleem

摘要

Glucan-type exopolysaccharides (EPSs) produced by lactic acid bacteria (LAB) have gained considerable attention owing to the increasing demand for natural resources in food and pharmaceutical industries. LAB use glucosyltransferases (GTFs) to synthesize glucan-type EPSs with different sizes and structures. Exploring new EPS-producing sources is key to discovering genes responsible for diverse EPS types. Recently, the complete genomes of two novel LAB species, Apilactobacillus waqarii HBW1 and Apilactobacillus iqraium F1 were reported. This study aims to identify and characterize EPS producing GTF genes in these species using computational approaches. RAST annotation revealed the presence of three EPS synthesizing genes each in A. waqarii (designated as AwGTF1, AwGTF2 and AwGTF3) and A. iqraium, (designated as GTFG1-F1, GTFG2-F1 and GTFG3-F1). Evolutionary analysis indicated that most of the retrieved GTFs shared low similarity with known GTFs and formed a distinct clade closely adjacent to dextran-producing L. mesenteroides, along with A. kunkeei. Domain analysis revealed a conserved domain architecture, though AwGTF3 lacks a glucan-binding domain (GBD). Due to the high molecular weight of GH70 family GTFs, 3D structure prediction was only possible using recently discovered computational tools, which revealed structural variations among these GTFs. Our findings suggest that out of six identified GTFs in A. waqarii and A. iqraium, five are putative glucansucrases while AwGTF3, contain a tryptophan residue in place of glycine, may function as a branching sucrase. Further studies involving recombinant gene expression and functional characterization could reveal detailed structure–function relationship of these enzymes.