02165nas a2200205 4500008004100000245010500041210006900146260000800215300001400223490000700237520142800244100002101672700002001693700002301713700002201736700001901758700002101777700003001798856013101828 2019 eng d00aMorphological and molecular characterization of Penicillium rubens sp.nov isolated from poultry feed0 aMorphological and molecular characterization of Penicillium rube cSEP a461–4780 v723 aFungi are the second largest group of eukaryotic organisms, estimates ranging from 2.2 to 3.8 million and 120,000 currently accepted species. Unfortunately, identification of Penicillium species is an overwhelming task even for taxonomic experts. In the current study, we isolated identified and morphologically characterized Penicillium rubens strain NCIM-1397. Molecular characterization of P. rubens by multilocus sequence analysis (MLSA) of internal transcribed spacer (ITS) region, β-tubulin (BenA), calmodulin (CaM), NL (D1/D2 region) and LR (large subunit region) gene. Conversely, BenA gene differentiated P. rubens from P. chrysogenum. Moreover, Vitek MS, Matrix-assisted laser desorption ionization–time of flight mass spectrometer (MALDI-TOF–MS) failed to discriminate to strain level. Metabolic profiling of P. rubens enable to produce roquefortine C, chrysogine, meleagrine, sorbicillin and non-producers for industrially important metabolite, penicillin determined by LC–MS. Even, In addition, even molecular detection of penicillin biosynthetic pathway genes, PcbC, PenDE and Phl were negative by PCR. Nevertheless, P. rubens showed 90–100% toxicity/pathogenicity on seed germination of sorghum (Sorghum bicolor) and no further development of coleoptiles, radicle and leaf length observed. Also, it showed non-toxic to animal cell determined on Human foreskin fibroblast (HFF) cell line by In-Vitro.1 aSawant, Amol, M.1 aVankudoth, Ramu1 aNavale, Vishwambar1 aKumavat, Rajkamal1 aKumari, Prabla1 aSanthakumari, B.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/morphological-and-molecular-characterization-penicillium-rubens-spnov-isolated-poultry-feed02147nas a2200193 4500008004100000022001400041245014800055210006900203260000900272300001600281490000600297520140000303100002101703700002901724700003001753700002601783700002001809856012401829 2020 eng d a2470-134300aProcess development for 6-aminopenicillanic acid production using lentikats-encapsulated escherichia coli cells expressing penicillin V acylase0 aProcess development for 6aminopenicillanic acid production using cNOV  a28972-289760 v53 a

Penicillin V acylase (PVA, EC 3.5.1.11) hydrolyzes the side chain of phenoxymethylpenicillin (Pen V) and finds application in the manufacture of the pharmaceutical intermediate 6-aminopenicillanic acid (6-APA). Here, we report the scale-up of cultivation of Escherichia coli whole cells expressing a highly active PVA from Pectobacterium atrosepticum and their encapsulation in polyvinyl alcohol-poly(ethylene glycol) Lentikats hydrogels. A biocatalytic process for the hydrolysis of 2% (w/v) Pen V was set up in a 2 L reactor using the Lentikats-immobilized whole cells, with a customized setup to enable continuous downstream processing of the reaction products. The biocatalytic reaction afforded complete conversion of Pen V for 10 reaction cycles, with an overall 90% conversion up to 50 cycles. The bioprocess was further scaled up to the pilot-scale at 10 L, enabling complete conversion of Pen V to 6-APA for 10 cycles. The 6-APA and phenoxy acetic acid products were recovered from downstream processing with isolated yields of 85-90 and 87-92%, respectively. Immobilization in Lentikats beads improved the stability of the whole cells on storage, maintaining 90-100% activity and similar conversion efficiency after 3 months at 4 degrees C. The robust PVA biocatalyst can be employed in a continuous process to provide a sustainable route for bulk 6-APA production from Pen V.

1 aSawant, Amol, M.1 aSunder, Avinash, Vellore1 aVamkudoth, Koteswara, Rao1 aRamasamy, Sureshkumar1 aPundle, Archana uhttps://library.ncl.res.in/content/process-development-6-aminopenicillanic-acid-production-using-lentikats-encapsulated02035nas a2200241 4500008004100000245012600041210006900167260000800236300000800244490000700252520115000259653002501409653002901434653002701463653003101490653001401521653002801535100002701563700002101590700002101611700003001632856013101662 2022 eng d00aAssembly, annotation, and comparative whole genome sequence of fusarium verticillioides isolated from stored maize grains0 aAssembly annotation and comparative whole genome sequence of fus cJUL a8100 v113 a

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' evolutionary history. The overall study helps in disease prevention and management of mycotoxins to ensure food safety.

10acomparative genomics10aFusarium verticillioides10amycotoxin biosynthesis10aplant-pathogen interaction10aSecretome10awhole-genome sequencing1 aNavale, Vishwambar, D.1 aSawant, Amol, M.1 aGowda, Varun, U.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/assembly-annotation-and-comparative-whole-genome-sequence-fusarium-verticillioides-isolated01994nas a2200217 4500008004100000022001400041245009600055210006900151260000800220300001200228490000700240520121800247653002701465653003901492653002401531653001501555653002801570100002101598700003001619856012701649 2022 eng d a0141-549200aBiosynthetic process and strain improvement approaches for industrial penicillin production0 aBiosynthetic process and strain improvement approaches for indus cFEB a179-1920 v443 a

Penicillins and cephalosporins are the most important class of beta (beta) lactam antibiotics, accounting for 65% total antibiotic market. Penicillins are produced by Penicillium rubens (popularly known as P. chrysogenum) were used to synthesize the active pharmaceutical intermediate (API), 6-aminopenicillinic acid (6-APA) employed in semisynthetic antibiotic production. The wild strains produce a negligible amount of penicillin (Pen). High antibiotic titre-producing P. chrysogenum strains are necessitating for industrial Pen production to meet global demand at lower prices. Classical strain improvement (CSI) approaches such as random mutagenesis, medium engineering, and fermentation are the cornerstones for high-titer Pen production. Since, Sir Alexander Fleming Discovery of Pen, great efforts are expanded to develop at a commercial scale antibiotics producing strains. Breakthroughs in genetic engineering, heterologous expression and CRISPR/Cas9 genome editing tools opened a new window for Pen production at a commercial scale to assure health crisis. The current state of knowledge, limitations of CSI and genetic engineering approaches to Pen production are discussed in this review.

10abeta-lactam antibiotic10aClassical strain improvement (CSI)10aGenetic engineering10aPenicillin10aPenicillium chrysogenum1 aSawant, Amol, M.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/biosynthetic-process-and-strain-improvement-approaches-industrial-penicillin-production02468nas a2200229 4500008004100000022001400041245009600055210006900151260000800220300001400228490000700242520169800249653001401947653001601961653002901977653001402006653001802020100002702038700002102065700003002086856012202116 2023 eng d a1415-475700aGenetic diversity of toxigenic Fusarium verticillioides associated with maize grains, India0 aGenetic diversity of toxigenic Fusarium verticillioides associat cMAY ae202200730 v463 a

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.

10adiversity10aFood safety10aFusarium verticillioides10aMycotoxin10apathogenicity1 aNavale, Vishwambar, D.1 aSawant, Amol, M.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/genetic-diversity-toxigenic-fusarium-verticillioides-associated-maize-grains-india02851nas a2200217 4500008004100000245013500041210006900176260000800245300000900253490000700262520199000269653002202259653004302281653003902324653003502363653002902398100002102427700002702448700003002475856012802505 2023 eng d00aIsolation and molecular characterization of indigenous penicillium chrysogenum/rubens strain portfolio for penicillin V production0 aIsolation and molecular characterization of indigenous penicilli cAPR a11320 v113 a

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.

10abeta-tubulin gene10ahigh-performance liquid chromatography10ainternal transcribed spacer region10aPenicillium chrysogenum/rubens10aphenoxymethyl penicillin1 aSawant, Amol, M.1 aNavale, Vishwambar, D.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/isolation-and-molecular-characterization-indigenous-penicillium-chrysogenumrubens-strain03188nas a2200241 4500008004100000022001400041245011800055210006900173260000800242300001400250490000700264520231600271653002502587653002102612653002802633653002302661653002602684653002802710100002102738700002702759700003002786856013002816 2024 eng d a1139-670900aGenome sequencing and analysis of penicillin V producing Penicillium rubens strain BIONCL P45 isolated from India0 aGenome sequencing and analysis of penicillin V producing Penicil cOCT a1473-14840 v273 a

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'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.

10acomparative genomics10aIllumina NovoSeq10aPenicillin biosynthesis10aPenicillium rubens10aSecondary metabolites10awhole genome sequencing1 aSawant, Amol, M.1 aNavale, Vishwambar, D.1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/genome-sequencing-and-analysis-penicillin-v-producing-penicillium-rubens-strain-bioncl-p45