Metabolic engineering for a clean environment: cell factories for removal and valorisation of heavy metal wastes

Heavy metal pollution significantly threatens environmental and human health, necessitating innovative remediation strategies. Bioremediation, employing organisms engineered to accumulate and transform contaminants, offers a promising, eco-friendly solution. Cell4NanoMet project aims to utilize metabolic engineering to develop organisms capable of concurrently removing heavy metals and converting them into valuable products, such as nanoparticles and nanoalloys.

Main domain

Food, bioeconomy, natural resources, biodiversity, agriculture and environment (ASC); Bioeconomy (DSIN); Environment and Eco-technologies (DSIN)

Main subdomain

Biodiversity recovery, conservation and sustainable restoration of ecosystems and ecosystem services

Secondary domain

Food, bioeconomy, natural resources, biodiversity, agriculture and environment (ASC); Bioeconomy (DSIN); Environment and Eco-technologies (DSIN)

Secondary subdomain

Circular bioeconomy

Tertiary domain

Food, bioeconomy, natural resources, biodiversity, agriculture and environment (ASC); Bioeconomy (DSIN); Environment and Eco-technologies (DSIN)

Tertiary subdomain

Water resources management and sustainable development of fisheries and aquaculture

Coordinator

University of Bucharest (UB)

Project director: Dr. Lavinia Ruta

Partner

APEL LASER S.R.L.

Objective 1

Designing efficient cell factories using yeast-based techniques to accumulate heavy metals and produce metal nanoparticles (MeNPs) via Yeast Molecular Display Technology.

Objective 2

Assessing the toxic effects of MeNPs on cells and identifying natural antioxidants to mitigate toxicity, leveraging Saccharomyces cerevisiae as a model organism.

Objective 3

Scaling up the developed techniques to pilot scale and employ natural antioxidants to enhance the life span of MeNP-exposed cell factories. This multidisciplinary approach holds promise for sustainable remediation and resource recovery from heavy metal pollution.

Results

Deliverables achieved:

Construction of pYD1-GFP: the expression vector was obtained and verified. Initial tests confirmed GFP localization on the cell surface.

Recombinant plasmids pYD1-GFP-MeBPep: generated for various heavy metal–binding peptides (His₆–His₁₀, Cys₆–Cys₁₀, Asp₆–Asp₁₀, Glu₆–Glu₁₀, Asp/Glu and Cys/Glu combinations).

Transformation of Saccharomyces cerevisiae cell lines: transformed colonies were obtained and confirmed by selection on specific media.

Validation of GFP expression: fluorescence microscopy confirmed surface expression of the constructs.

Result indicators:

Obtaining two artificial sequences enriched in amino acids with affinity for metal ions.

Confirmation of recombinant plasmids by electrophoresis and restriction enzyme analysis.

GFP expression under the control of the GAL1 promoter, visualized by fluorescence.

Establishment of a proof of concept for the “Yeast Surface Display” technology applied to metal-binding peptides.

Dissemination of results:

Research results were presented at the Yeast in Bioeconomy Conference, October 22–24, 2025, Compiègne, France, PO15. Yeast surface display as a sustainable strategy for heavy metal removal from water – poster by Lavinia Ruta, Andrei Barboi, Ileana Farcasanu.