ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Ellappan Vanitha and Krishnan Dhandayuthapani
PG and Research Department of Botany, Arignar Anna Government Arts College, Cheyyar (Affiliated to Thiruvalluva University, Vellore), Tamil Nadu, India.
Article Number: 10688 | © The Author(s). 2025
J Pure Appl Microbiol. 2025;19(4):2850-2863. https://doi.org/10.22207/JPAM.19.4.22
Received: 15 June 2025 | Accepted: 08 September 2025 | Published online: 13 November 2025
Issue online: December 2025
Abstract

This study presents an integrated approach for solvent free recovery of total phenolic compounds (TPC) from the biomass of double mutant microalga Chlorococcum humicola MCH4 using microwave-assisted pretreatment (MAPT) followed by submerged fermentation (SmF) with Bacillus subtilis. This microalga strain was developed through UV-C and EMS-induced mutagenesis and cultivated in ultrasonically pretreated municipal wastewater to obtain biomass, which was then subjected to MAPT to improve the release of TPC. Key fermentation conditions such as temperature, pH, fermentation time, agitation speed and zinc oxide nanoparticles (ZnO-NPs) concentration were optimized to maximize TPC yield. Under optimal conditions-microwave power of 600 W, pH 7.0, temperature 35 °C, agitation at 150 rpm, fermentation time of 48 h, and ZnO-NPs supplementation at 150 mgL-1, the process achieved a maximum TPC yield of 60.72 ± 0.11 mg GAE g-1 DW. The IC50 values of 38.7 µgmL-1 and 41.2 µgmL-1 were obtained for antioxidant activity of the recovered TPC by DPPH and ABTS assay, respectively, indication strong antioxidant potential. This work highlights the synergistic effect of MAPT and bacterial fermentation for effective solvent free recovery of bioactive phenolics from microalgal biomass. The process is both scalable and environmentally sustainable, indicating robust possible for use in the nutraceutical and pharmaceutical industries.

Keywords

Antioxidant Activity, Chlorococcum humicola, Microwave-assisted Pretreatment, Submerged Fermentation, Total Phenolic Compounds

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© The Author(s) 2025. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.