@article {48093, title = {Assembly, annotation, and comparative whole genome sequence of fusarium verticillioides isolated from stored maize grains}, volume = {11}, year = {2022}, month = {JUL}, pages = {810}, type = {Article}, abstract = {

Fusarium verticillioides is a plant pathogenic fungus affecting a wide range of crops worldwide due to its toxigenic properties. F. verticillioides BIONCL4 strain was isolated from stored maize grain samples in India, and produces high amount of fumonisin B1 (FB1). We report a comparative genomic analysis of F. verticillioides, covering the basic genome information, secretome, and proteins involved in host-pathogen interactions and mycotoxin biosynthesis. Whole-genome sequencing (WGS) was performed using the Illumina platform with an assembly size of 42.91 Mb, GC content of 48.24\%, and 98.50\% coverage with the reference genome (GCA000149555). It encodes 15,053 proteins, including 2058 secretory proteins, 676 classical secretory proteins, and 569 virulence and pathogenicity-related proteins. There were also 1447 genes linked to carbohydrate active enzymes (CaZymes) and 167 genes related to mycotoxin production. Furthermore, F. verticillioides genome comparison revealed information about the species{\textquoteright} evolutionary history. The overall study helps in disease prevention and management of mycotoxins to ensure food safety.

}, keywords = {comparative genomics, Fusarium verticillioides, mycotoxin biosynthesis, plant-pathogen interaction, Secretome, whole-genome sequencing}, doi = {10.3390/pathogens11070810}, author = {Navale, Vishwambar D. and Sawant, Amol M. and Gowda, Varun U. and Vamkudoth, Koteswara Rao} } @article {48035, title = {Toxicity and preventive approaches of Fusarium derived mycotoxins using lactic acid bacteria: state of the art}, journal = {Biotechnology Letters}, volume = {44}, year = {2022}, month = {OCT}, pages = {1111-1126}, type = {Review}, abstract = {

Mycotoxin contamination of food and feed is a serious food safety issue and causes acute and chronic diseases in humans and livestock. Climatic and agronomic changes helps in the proliferation of fungal growth and mycotoxin production in food commodities. Mycotoxin contamination has attracted global attention due to its wide range of toxicity to humans and animals. However, physical and chemical management approaches in practice are unsafe for well-being due to their health-hazardous nature. Various antibiotics and preservatives are in use to reduce the microbial load and improve the shelf life of food products. In addition, the use of antibiotic growth promotors in livestock production may increase the risk of antimicrobial resistance, which is a global health concern. Due to their many uses, probiotics are helpful microbes that have a significant impact on food and nutrition. Furthermore, the probiotic potential of lactic acid bacteria (LAB) is employed in various food and feed preparations to neutralize mycotoxins, antimicrobial activities, balance the gut microbiome, and various immunomodulatory activities in both humans and livestock. In addition, LAB produces various antimicrobials, flavouring agents, peptides, and proteins linked to various food and health care applications. The LAB-based processes for mycotoxin management are more effective, eco-friendly, and low-cost than physical and chemical approaches. The toxicity, novel preventive measures, binding nature, and molecular mechanisms of mycotoxins{\textquoteright} detoxification using LAB have been highlighted in this review.

}, keywords = {Food safety, Fusarium species, Lactic acid bacteria, management, Mycotoxin}, issn = {0141-5492}, doi = {10.1007/s10529-022-03293-4}, author = {Navale, Vishwambar D. and Vamkudoth, Koteswara Rao} } @article {48298, title = {Genetic diversity of toxigenic Fusarium verticillioides associated with maize grains, India}, journal = {Genetics and Molecular Biology}, volume = {46}, year = {2023}, month = {MAY}, pages = {e20220073}, type = {Article}, abstract = {

In the present investigation, prevalence, genetic diversity, and mycotoxin producing potential of Fusarium species associated with maize grain samples were studied from different geographical regions of India. The highest prevalence of Fusarium verticillioides was recorded as 88.52\%, followed by F. coffeatum, F. foetens, and F. euwallaceae, 6.55\%, 3.27\%, and 1.63\%, respectively. We isolated 54 strains of F. verticillioides, and their genetic diversity was studied by inter simple sequence repeats (ISSR). The ISSR fingerprints (AG) 8C and (AG) 8G showed 252 and 368 microsatellite sites in the genome of F. verticillioides and resulted in 99-100\% repeatability and reproducibility. The Simpson (SID) and Shannon (H) indices (0.78 and 2.36) suggest that F. verticillioides strains exhibit moderate to high diversity. Molecular detection of fumonisin B1 (FB1) biosynthetic genes (FUM1 and FUM13) involved in FB1 production in F. verticillioides was confirmed by polymerase chain reaction (PCR). Furthermore, 91\% of the strains were positive for FB1 production, which was affirmed by liquid chromatography with tandem mass spectrometry (LC-MS-MS). In-vitro appurtenance of F. verticillioides spores exhibited a high to moderate effect on the growth and development of the maize. The current finding demonstrated that most F. verticillioides strains showed a wide range of genetic diversity with varied toxigenic and pathogenic potentials. In conclusion, for the first time, F. coffeatum, F. foetens, and F. euwallaceae species were reported from maize grain samples in India. They were positive for FB1 and negatively affecting grain quality, which is a major concern in food safety.

}, keywords = {diversity, Food safety, Fusarium verticillioides, Mycotoxin, pathogenicity}, issn = {1415-4757}, doi = {10.1590/1678-4685-GMB-2022-0073}, author = {Navale, Vishwambar D. and Sawant, Amol M. and Vamkudoth, Koteswara Rao} } @article {48717, title = {Isolation and molecular characterization of indigenous penicillium chrysogenum/rubens strain portfolio for penicillin V production}, journal = {Microorganisms}, volume = {11}, year = {2023}, month = {APR}, pages = {1132}, type = {Article}, abstract = {

Beta (beta)-lactam antibiotic is an industrially important molecule produced by Penicillium chrysogenum/rubens. Penicillin is a building block for 6-aminopenicillanic acid (6-APA), an important active pharmaceutical intermediate (API) used for semi-synthetic antibiotics biosynthesis. In this investigation, we isolated and identified Penicillium chrysogenum, P. rubens, P. brocae, P. citrinum, Aspergillus fumigatus, A. sydowii, Talaromyces tratensis, Scopulariopsis brevicaulis, P. oxalicum, and P. dipodomyicola using the internal transcribed spacer (ITS) region and the beta-tubulin (BenA) gene for precise species identification from Indian origin. Furthermore, the BenA gene distinguished between complex species of P. chrysogenum and P. rubens to a certain extent which partially failed by the ITS region. In addition, these species were distinguished by metabolic markers profiled by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Secalonic acid, Meleagrin, and Roquefortine C were absent in P. rubens. The crude extract evaluated for PenV production by antibacterial activities by well diffusion method against Staphylococcus aureus NCIM-2079. A high-performance liquid chromatography (HPLC) method was developed for simultaneous detection of 6-APA, phenoxymethyl penicillin (PenV), and phenoxyacetic acid (POA). The pivotal objective was the development of an indigenous strain portfolio for PenV production. Here, a library of 80 strains of P. chrysogenum/rubens was screened for PenV production. Results showed 28 strains capable of producing PenV in a range from 10 to 120 mg/L when 80 strains were screened for its production. In addition, fermentation parameters, precursor concentration, incubation period, inoculum size, pH, and temperature were monitored for the improved PenV production using promising P. rubens strain BIONCL P45. In conclusion, P. chrysogenum/rubens strains can be explored for the industrial-scale PenV production.

}, keywords = {beta-tubulin gene, high-performance liquid chromatography, internal transcribed spacer region, Penicillium chrysogenum/rubens, phenoxymethyl penicillin}, doi = {10.3390/microorganisms11051132}, author = {Sawant, Amol M. and Navale, Vishwambar D. and Vamkudoth, Koteswara Rao} } @article {48733, title = {Metabolites from lactococcus lactis subsp. lactis: isolation, structure elucidation, and antimicrobial activity}, journal = {ACS Omega}, volume = {8}, year = {2023}, month = {SEP}, pages = {36628-36635}, type = {Article}, abstract = {

Herein, we disclose the identification of novel metabolites from a potential probiotic strain, Lactococcus lactis subsp. lactis, obtained from traditional dairy milk samples collected in Maharashtra, India (in January 2021). Isolated metabolites include pyrazin-2-carboxamide [1, pyrazinamide, a potential antitubercular drug], 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (2, DDMP), 2,4-di-tert-butylphenol (3), and hexadecanoic acid (4, palmitic acid). The chemical structures of these metabolites were elucidated through extensive 1D NMR (H-1 and C-13) and 2D NMR (HSQC, HMBC, and NOESY) analyses, high-resolution mass spectrometry, high-performance liquid chromatography, and single-crystal X-ray crystallography. Furthermore, these novel metabolites exhibited potent inhibitory activities against various bacteria, fungi, and yeast strains with minimum inhibitory concentrations ranging between 1.56 and 25 mu g/mL, and compounds 1 and 3 were found to be most active against a wide range of microbial strains tested.

}, issn = {2470-1343}, doi = {10.1021/acsomega.3c01662}, author = {Navale, Vishwambar D. and Borade, Balasaheb R. and Rama Krishna, Gamidi and Vamkudoth, Koteswara Rao and Kontham, Ravindar} } @article {49325, title = {Genome sequencing and analysis of penicillin V producing Penicillium rubens strain BIONCL P45 isolated from India}, journal = {International Microbiology}, volume = {27}, year = {2024}, month = {OCT}, pages = {1473-1484}, type = {Article}, abstract = {

Background A filamentous fungus Penicillium rubens is widely recognized for producing industrially important antibiotic, penicillin at industrial scale. Objective To better comprehend, the genetic blueprint of the wild-type P. rubens was isolated from India to identify the genetic/biosynthetic pathways for phenoxymethylpenicillin (penicillin V, PenV) and other secondary metabolites. Method Genomic DNA (gDNA) was isolated, and library was prepared as per Illumina platform. Whole genome sequencing (WGS) was performed according to Illumina NovoSeq platform. Further, SOAPdenovo was used to assemble the short reads validated by Bowtie-2 and SAMtools packages. Glimmer and GeneMark were used to dig out total genes in genome. Functional annotation of predicted proteins was performed by NCBI non-redundant (NR), UniProt, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO) databases. Moreover, secretome analysis was performed by SignalP 4.1 and TargetP v1.1 and carbohydrate-active enzymes (CAZymes) and protease families by CAZy database. Comparative genome analysis was performed by Mauve 2.4.0. software to find genomic correlation between P. rubens BIONCL P45 and Penicillium chrysogenum Wisconsin 54-1255; also phylogeny was prepared with known penicillin producing strains by ParSNP tool. Results Penicillium rubens BIONCL P45 strain was isolated from India and is producing excess PenV. The 31.09 Mb genome was assembled with 95.6\% coverage of the reference genome P. chrysogenum Wis 54-1255 with 10687 protein coding genes, 3502 genes had homologs in NR, UniProt, KEGG, and GO databases. Additionally, 358 CAZymes and 911 transporter coding genes were found in genome. Genome contains complete pathways for penicillin, homogentisate pathway of phenyl acetic acid (PAA) catabolism, Andrastin A, Sorbicillin, Roquefortine C, and Meleagrin. Comparative genome analysis of BIONCL P45 and Wis 54-1255 revealed 99.89\% coverage with 2952 common KEGG orthologous protein-coding genes. Phylogenetic analysis revealed that BIONCL P45 was clustered with Fleming{\textquoteright}s original isolate P. rubens IMI 15378. Conclusion This genome can be a helpful resource for further research in developing fermentation processes and strain engineering approaches for high titer penicillin production.

}, keywords = {comparative genomics, Illumina NovoSeq, Penicillin biosynthesis, Penicillium rubens, Secondary metabolites, whole genome sequencing}, issn = {1139-6709}, doi = {10.1007/s10123-024-00491-0}, author = {Sawant, Amol M. and Navale, Vishwambar D. and Vamkudoth, Koteswara Rao} } @article {49272, title = {Unlocking the potential of lactococcus lactis subsp. lactis BIONCL17752 strain on fumonisin B1 production by fusarium verticillioides}, journal = {Food Control}, volume = {168}, year = {2025}, month = {FEB}, pages = {110910}, type = {Article}, abstract = {

Mycotoxins are the potent toxic chemical agent{\textquoteright}s linked to food safety, pose a serious risk to human and animal health globally. The management of toxigenic fungi is a challenging task in food and feed industries. The present investigation aimed to use probiotic potentials of lactic acid bacteria (LAB), and fermented cell-free broth as antifungal agents and neutralization of carcinogenic mycotoxin, fumonisin B1 (FB1) in food and feed considered as {\textquoteleft}{\textquoteleft}turned waste into treasure{\textquoteright}{\textquoteright}. We strived to investigate the probiotic potential of Lactococcus lactis subsp. lactis BIONCL17752 strain on prevention of growth and neutralization of FB1 produced by Fusarium verticillioides BIONCL4 strain to emulate as as a food grade bio-preservative. The BIONCL17752 strain exhibited excellent inhibitory activity against BIONCL4 strain. It was cultivated in MRS medium, and obtained cells pellet (CP) and cell-free supernatant (CFS) was lyophilized, and used for their antifungal activity against BIONCL4 strain. The minimum fungicidal concentration (MFC) determined as 12.5 mu g/mL. Nevertheless, CFS assessed for neutralization of FB1 production which exhibited complete inhibition and downregulation of FB1 encoding FUM1 gene expression using quantitative real-time PCR (qPCR). Furthermore, CFS induced transcriptomic studies against BIONCL4 strain endorsed a significant downregulation of virulence, FB1, fusaric acid, fusarin, and chitin biosynthetic pathway genes. The CFS and cells reticent spore germination and FB1 production in the range of 40-61\% and 74-85\%, respectively under stored maize for 60 days. The current findings suggest BIONCL17752 strain apprehend the fungal growth and inhibit the FB1, and other toxigenic molecules biosynthesis, and can be employed as food grade preservative in the food industries to ensure the food safety and human health.

}, keywords = {Food grade preservative, Food safety, Fumonisin B-1, Fusarium verticillioides, lactis, Lactococcus lactis subsp., Maize}, issn = {0956-7135}, doi = {10.1016/j.foodcont.2024.110910}, author = {Navale, Vishwambar D. and Vamkudoth, Koteswara Rao} }