Project for production technology of glutathione-producing yeast for food technology
Students Name: Zubar Zoriana Orestivna
Qualification Level: magister
Speciality: Biotechnology and Bioengineering
Institute: Institute of Chemistry and Chemical Technologies
Mode of Study: full
Academic Year: 2025-2026 н.р.
Language of Defence: ukrainian
Abstract: Study object – glutathione-producing biomass of Saccharomyces cerevisiae and extraction utilizing pulsed electric field (PEF) electroporation. Scope of research – industrial biotechnology of microbial antioxidants; fer-mentation technology; non-thermal cell-disruption methods. Research objective – development, optimization, and validation of an inte-grated biotechnological process for production of food-grade reduced glutathione from yeast biomass, including fermentation, PEF-assisted cell disruption, extraction, purification, and evaluation of industrial applicability. Glutathione being a predominant intracellular antioxidant involved in redox regulation, detoxification, and metabolic homeostasis, it has gained an increasing application in food technology, nutraceuticals, pharmaceuticals, and functional in-gredients. Given the high rates of production of natural biologically active com-pounds, the pursuit of efficient and environmentally friendly production techniques is of interest. Conventional chemical and enzymatic approaches to glutathione syn-thesis face formidable drawbacks in terms of high costs, environmental impact, and biological activity limitations that have led industrial interest to rely on microbial fermentation. Saccharomyces cerevisiae is a safe and effective manufacturer with GRAS status, metabolic robustness, and intrinsic capacity to capture high concentrations of glutathione in an intracellular environment. In optimized fermentations the yeast biomass level reaches 1.8–2.0% GSH in the ratio of dry weight. But to extract bio-logically active reduced glutathione, it still faces significant challenge, since me-chanical or thermal perturbation of the molecule can damage or deposit protein con-tamination. Therefore, this paper aims to tackle this issue by means of pulsed elec-tric field (PEF) technology, a non-thermal electroporation method which promotes the permeabilization of yeast cells, enhances the release of GSH at appropriate tem-perature, reduces thermal degradation, and minimizes the energy consumption. The scientific novelty of this work is in combining high-density fed-batch yeast fermentation with a pilot-scale PEF electroporation process that is tailored to-wards the recovery of glutathione. This concept has not been previously employed in the Ukrainian bioprocessing practice and offers an advanced technology model on local glutathione cultivation, which is also valuable in view of the present situa-tion by Ukraine’s lack of industrial production of glutathione and the reliance on imports. The entire technical construction, including preparation of raw material, 2-step inoculum farming, high-rate fermentation, separation of biomass, PEF treat-ment, ultrafiltration and ion-exchange chromatography purification, and drying of product, were included within the thesis. A material balance was carried out at each production stage and the potential of producing 20 kg of purified glutathione annu-ally was verified. A detailed process flow diagram and equipment specification was submitted. Technical-economic parameters of the biotechnology-based program were determined in the economic section, to verify the technology capability and compet-itiveness of the developed biotechnology under pilot-run conditions. The proposed process reduces energy consumption, saves operational overhead compared to me-chanical disruption, and is congruent with the development of existing fermentation infrastructure. Conclusion is supported by literature review and experimental data with modern biochemical and technological assessment techniques. Key words: glutathione, Saccharomyces cerevisiae, pulsed electric field (PEF), fermentation technology, cell electroporation, bioprocess engineering.