Annotating Multiple Prebiotic Synthesizing Capabilities Through Whole Genome Sequencing of Fusarium Strain HFK-74

In the present study, seven fungal isolates from effluent treatment plants were screened for the production of prebiotic fructooligosaccharide synthesizing enzymes with the highest activity of fructofuranosidase (17.52 U/mL) and fructosyl transferase (18.92 U/mL) in strain HKF-74. Mining of genome sequence of strain revealed the annotation of genes providing multiple carbohydrate metabolizing capacities, such as amylases (AMY1), beta-galactosidase (BGAL), beta-xylosidase (Xyl), β-fructofuranosidase (ScrB), fructosyltransferase (FTF), and maltose hydrolases (malH). The annotated genes were further supported by β-galactosidase (15.90 U/mL), xylanase (17.91 U/mL), and α-amylase (14.05 U/mL) activities for synthesis of galactooligosaccharides, xylooligosaccarides, and maltooligosaccharides, respectively. In addition to genes encoding prebiotic synthesizing enzymes, four biosynthetic gene clusters (BGCs) including Type I polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS), NRPS-like, and terpene were also predicted in strain HKF-74. This was significant considering their potential role in pharmaceutical and therapeutic applications as well as in virulence. Accurate taxonomic assignment of strain HKF-74 by in silico genomic comparison indicated its closest identity to type strains Fusarium verticillioides NRRL 20984, and 7600. The average nucleotide identity (ANI) of strain HKF-74 with these strains was 92.5% which was close to the species threshold cut-off value (95-96%) while the DNA-DNA hybridization (DDH) value was 83-84% which was greater than both, species delineating (79-80%), and also sub-species delineating (70%) boundaries. Our findings provide a foundation for further research into the use of Fusarium strains and their prebiotic synthesizing enzymes for the development of novel prebiotic supplements.