Isolation and Molecular Characterization of Indigenous Penicillium chrysogenum/rubens Strain Portfolio for Penicillin V Production

Abstract

Beta (β)-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 β-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.

An Adsorption Based Downstream Processing Approach for Penicillin V from a Penicillium chrysogenum BIONCL I22 Culture Filtrate

“…To compare solvent extraction with the resin-based recovery method, butyl acetate-based extraction of penicillin V was initially demonstrated using a 1 mg/mL solution of standard penicillin V followed by extraction from the culture filtrate. It was first extracted into n -butyl acetate (pH 2.5), followed by back extraction into phosphate buffer (pH 7.5) ,, (Figure S2). The effect of varying volume ratios of penicillin V solution to the n -butyl acetate organic phase was studied (Figure S3 and Table S2).…”

“…Furthermore, it was cultured for 8 days at 25 °C and 180 rpm in a rotary incubator shaker. Following incubation, the culture was harvested and initially filtered with a muslin cloth, followed by filtering with a 0.22 μm filter. ,, …”

“…Following incubation, the culture was harvested and initially filtered with a muslin cloth, followed by filtering with a 0.22 μm filter. 19 , 57 , 58 …”

“…Following incubation, the culture was harvested and initially filtered with a muslin cloth, followed by filtering with a 0.22 μm filter. 19,57,58 4.9. Batch Adsorption Experiments from the P. chrysogenum BIONCL I22 Culture Filtrate.…”

“…Penicillin V being one such essential antibiotic, the development of a novel, effective, and environmentally friendly extraction process would be quite advantageous. Conventional downstream processing of penicillin V using solvent extraction is a multistep procedure, in which, it is first extracted into an organic solvent using n -butyl acetate or amyl acetate, followed by back extraction into phosphate buffer (PBS). , This method has various drawbacks, such as high solvent consumption, multiple extraction steps, less yield, and emulsification. Apart from butyl acetate-based extraction, numerous alternative approaches have been developed to extract penicillins.…”