Production of phospholipase C by Pseudomonas fluorescens MICAYA 2023

Document Type : Original Article


Agricultural Microbiology Department, Faculty of Agriculture, Ain Shams University, Cairo, Egypt


Phospholipases represent versatile biocatalysts that are widely used in various industrial applications, Such as for biodiesels, food, oil degumming, and detergents. The purpose of this work was to use design expert to enhance the production of phospholipase C (PLC) from the most efficient strain, which can then be applied to degumming. PLC producing bacteria were isolated from Egyptian soil and food sources. A potent Pseudomonas isolate was identified as P. fluorescens MICAYA 2023 by 16S rRNA sequencing and submitted in the GenBank with accession number (OQ231499). Optimization via Plackett Burman and central composite design found soybean meal, yeast extract, NaCl and egg yolk significantly enhanced phospholipase C production. Michaelis-Menten kinetics determined a Km of 0.4 mg/ml egg yolk and Vmax 287 U/ml. Box Behnken design identified optimal pH 6.5, 0.55 g/l CaCO3, 1.05% egg yolk, 48.5°C for 395 U/ml phospholipase C production. The phospholipase showed low cytotoxicity against human fibroblast cells. PLC concentrations (0.2-1 ml) effectively degummed Sesamum indicum, Matricaria chamomilla, Nasturtium officinale, Simmondsia chinensis, Olea europaea, Nigella , and Ricinus communis oils. PLC concentrations of 0.4-0.8 ml/L yielded the highest phospholipid reduction. P. fluorescens phospholipase C offers a biodegradable alternative to chemical degumming. Statistical optimization successfully increased phospholipase C yields. Characterization found the enzyme works best at alkaline pH, moderate temperature, and with egg yolk substrate. The bio-degumming capability was demonstrated for multiple plant seed oils. Further immobilization and protein engineering could enhance the industrial applicability of PLC.


Main Subjects

Articles in Press, Accepted Manuscript
Available Online from 07 May 2024
  • Receive Date: 17 March 2024
  • Revise Date: 03 May 2024
  • Accept Date: 07 May 2024