imagine (Institute of molecular genetics and genetic engineering)
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Dinuclear Gallium(III) Complex With 1,3-Propanediamine-N,N′-Diacetate: Structural Characterization, Antimicrobial Activity, and DNA/BSA Interactions
No full textIn this study, a tetradentate 1,3-propanediamine-N,N′-diacetate (1,3-pdda2−) was utilized for the synthesis of a dinuclear gallium(III) complex, uns-cis-[Ga(1,3-pdda)(µ-OH)]2.2H2O (1). Complex 1 was characterized using IR and NMR (1H and 13C) spectroscopy, and its crystal structure was determined by single-crystal X-ray diffraction analysis. Both Ga(III) ions in Complex 1 exhibit octahedral geometry, with each ion coordinated by two nitrogen and two oxygen atoms from the 1,3-pdda2– ligand, as well as two oxygen atoms from the bridging hydroxyl groups. IR and NMR (1H and 13C) spectra were simulated using DFT methods, showing a high degree of correlation with experimental data. Hirshfeld surface analysis provided insights into intermolecular interactions, with H⋯O and H⋯H interactions contributing significantly to the crystal stability. The antimicrobial potential of Complex 1 was evaluated alongside previously synthesized gallium(III) complexes, Na[Ga(1,3-pdta)]·3H2O (2) and Ba[Ga(1,3-pndta)]2·3H2O (3), with 1,3-pdta4− (1,3-propanediamine-N,N,N′,N′-tetraacetate) and 1,3-pndta4− ((±)-1,3-pentanediamine-N,N,N′,N′-tetraacetate), respectively. Among all the tested microbial species, the gallium(III) complexes have shown selective activity against Pseudomonas aeruginosa PAO1 strain and were able to reduce pyocyanin production by 40–43% in the clinical isolate BK25H of this bacterium. Moreover, Complexes 1–3 can modulate the quinolone-mediated quorum sensing system in P. aeruginosa PAO1. Interaction studies with calf thymus DNA (ct-DNA) and bovine serum albumin (BSA) were conducted to evaluate the binding affinity and mode of interaction of Complex 1 with key biomolecules, aiming to assess its potential for transport via serum proteins and its safety profile in terms of DNA interactions. Spectrofluorimetric experiments and molecular docking revealed that Complex 1 binds strongly to the Site I on BSA, with weaker interactions at the Site II. While spectrofluorimetric studies showed that Complex 1 has a slight affinity for minor groove binding or intercalation to ct-DNA, docking studies suggested some minor groove binding, especially in larger DNA sequences, with enhanced stabilization in 10-bp-DNA through hydrogen and carbon bonds
Improving enzymatic degradation of unpretreated poly(ethylene terephthalate)
No full textAlthough the efficiency of poly(ethylene terephthalate) (PET) degradation has been successfully improved by depolymerase engineering, mostly by using Goodfellow-PET (gf-PET) as a substrate, efforts to degrade unpretreated PET materials with high crystallinity remain insufficient. Here, we endeavored to improve the degradation capability of a WCCG mutant of leaf-branch compost cutinase (LCC) on a unpretreated PET substrate (crystallinity > 40%) by employing iterative saturation mutagenesis. Using this method, we developed a high-throughput screening strategy appropriate for unpretreated substrates. Through extensive screening of residues around the substrate-binding groove, two variants, WCCG-sup1 and WCCG-sup2, showed good depolymerization capabilities with both high- (42%) and low-crystallinity (9%) substrates. The WCCG-sup1 variant completely depolymerized a commercial unpretreated PET product in 36 h at 72 °C. In addition to enzyme thermostability and catalytic efficiency, the adsorption of enzymes onto substrates plays an important role in PET degradation. This study provides valuable insights into the structure-function relationship of LCC
THE GOLDEN JACKAL (CANIS AUREUS) AS A RESERVOIR AND VECTOR FOR ECHINOCOCCUS SPP.
No full textThe golden jackal (Canis aureus) serves as a definitive and occasionally, an intermediate host for
Echinococcus parasites. Serbia hosts one of the largest golden jackal populations in Europe, and
represents a key region for studying the ecological role of the jackal in the maintenance and spread
of Echinococcus spp. To assess the occurrence of Echinococcus spp. in golden jackals, we
inspected the gastrointestinal tract of legally hunted animals (n=101) from different areas of
Serbia. The intestines were frozen for several weeks prior to examination to inactivate
Echinococcus spp. eggs. The fecal samples were collected and Taeniid eggs were purified by zinc
chloride (ZnCl₂) flotation and sequential mesh filtration. Total DNA (deoxyribonucleic acid) was
extracted and multiplex PCR (polymerase chain reaction) for the detection of Echinococcus
multilocularis, Echinococcus granulosus and Echinococcus canadensis was performed. The
occurrence of Echinococcus spp. was found to be 7.9%, with Echinococcus multilocularis detected
in seven golden jackals, and Echinococcus canadensis in one. Notably, four infected animals
originated from eastern Serbia, an area that may be a hotspot for transmission and warrants further
monitoring. Of particular concern to public health however, is that Echinococcus multilocularis
was present in two infected animals (out of six examined) from a suburb of Niš, the third most
populated city in Serbia. The results of this study emphasize the role of golden jackals as important
reservoirs and vectors for Echinococcus tapeworms in natural ecosystems. In addition, the
increasing expansion of jackal populations into peri-urban environments raises the risk of zoonotic
transmission to humans.Proceedings: Teslić, Banja Vrućica, June, 11th–14th, 2025
Degradation of biomaterials by Streptomyces microflavus DG19: depolymerization activity, genome mining, and soil burial assessment
No full textThe accumulation of plastic waste remains a significant environmental challenge despite the alarming evidence and public efforts, emphasizing the need for biodegradable alternatives and appropriate remediation strategies. In this study, Streptomyces microflavus DG19 was evaluated for capacity to degrade a selection of biomaterials that are increasingly penetrating market as readily degradable alternatives. S. microflavus DG19 rapidly degraded poly(3-hydroxybutyrate-co-3-hydroxyvalerate) films in liquid culture (96% weight loss in 7 days) and demonstrated activity against poly(ε-caprolactone) in both agar-based and liquid culture experiments and against cellulose in Congo red assay. 3-Hydroxybutyrate and lactic acid were also metabolized. Genomic analysis identified a number of enzymes involved in carbohydrate and bioplastic degradation. A putative extracellular poly(3-hydroxybutyrate) (PHB) depolymerase (SmPHBase) containing a variant substrate binding domain, and other enzymes involved in 3-hydroxybutyrate metabolism, were of special interest. The presence of > 30 biosynthetic gene clusters highlights this strain’s potential for upcycling bioplastic-containing waste. Soil burial tests demonstrated substantial weight loss in pure biomaterial films and multilayer consumable items containing PHB, showcasing the applicability of S. microflavus DG19 as a composting enhancer. Overall, the findings highlight the pertinence of specialized bacterial strains to biomaterial recycling and upcycling
PET as bacterial carbon source - a new hope for plastic pollution
No full textPolyethylene terephthalate (PET), one of the most widely used petrochemical-based thermoplastics, can have significant harmful environmental and health impacts if not disposed of properly due to its recalcitrancy to degradation under environmental conditions. Moreover, despite the available technologies for its recycling, many wasted PET products comprise impurities of different nature (e.g., paper, oils, ink, etc.), additives or blends with other polymers, that make their recycling impractical. Therefore, novel approaches are still required to complement the existing ones.
In line with this, the present work reports the valorisation of post-consumer PET waste into value-added microbial biomass. Post-consumer PET waste, containing approximately 95% PET, 5% polyethylene (PE), <1% carbon black additives, was depolymerized via reactive extrusion, resulting in a material named REX-PET. This material was processed into an aqueous solution containing sodium terephthalate (TPA) (REX-TPA solution) that was used as the sole carbon source for the cultivation of Glutamicibacter nicotianae strain 24A, isolated from soil contaminated with plastic waste collected at a municipal solid waste management facility. The fed-batch bioreactor cultivation of G. nicotianae strain 24A, under controlled temperature (30 ºC) and pH (7.0), resulted in the production of 9.4 g/L of dry biomass, with a protein content of 48 wt%, within 62 h of cultivation. A TPA consumption rate of 13.6 g/(L.day) was displayed by the culture that reached a TPA removal efficiency of 89% and a biomass yield of 0.4 g/g. Interestingly, the biomass presented a bright yellow coloration indicative of the presence of a pigment, which was extracted and characterized.
Therefore, this study demonstrates the promising potential of the newly isolated G. nicotianae strain 24A for the biological valorisation of post-consumer PET waste into biomass and pigments of high value, thus contributing for tackling the problems associated with the management of post-consumer PET waste contaminated with other chemicals, whose current recycling in not feasible.21st International Conference on
Renewable Resources
and Biorefineries
Moving Beyond Carbon Neutral
2 – 4 June 2025
Turku/Åbo, Finlan
BIOCATALYTIC PLASTIC DEGRADATION BY BIOFILM-FORMING PSEUDOMONAS STRAINS EXPRESSING CUTINASE
No full textPlastic materials such as polyethylene (PE), polyethylene terephthalate (PET), but also biodegradable poly (lactic
acid) (PLA) and polyhydroxyalkanoate (PHA), are widely used in various industries, but they are not easily
degradable, which significantly contributes to environmental pollution. Traditional waste management methods,
including landfilling, incineration, and mechanical or chemical recycling, have limited effectiveness, with less
than 10% of plastic waste being successfully recycled globally1. Therefore, innovative and environmentally safe
biotechnological approaches for sustainable end-of-life options are needed to address this issue.
Microbial biofilms, which are complex communities of microorganisms adhering to surfaces, play an important
role in various environmental processes, including biodegradation. Due to their enhanced resistance to
environmental stressors, such as chemical exposure and antimicrobial agents, biofilms serve as an ideal model
for exploring the biodegradation potential of microorganisms. Extracellular enzymes secreted by
microorganisms within biofilms have been shown to catalyze the breakdown of various recalcitrant substrates,
including polymers like plastics2. This study investigates the potential of biofilm-forming Pseudomonas
aeruginosa strains (BK25H, BR5H, and S25H), expressing leaf compost cutinase (LCC) from the pSEVA238
plasmid, to degrade plastic materials.
Wild-type P. aeruginosa strains and their constructs were first tested in a growth assay to determine if they could
grow on a substrate supplemented with plastic monomers as the sole carbon source (ethylene glycol, terephthalic
acid, lactic acid, 3-hydroxybutyric acid, and 3-hydroxy-octanoic acid). The results indicated that the P.
aeruginosa strains were unable to grow under these conditions, while LCC-expressing variants BK25H-LCC and
BR5H-LCC demonstrated growth on ethylene glycol. In a disc assay using minimal media supplemented with
various plastic polymers (PLA, PHA, polyhydroxybutyrate (PHB), polyhydroxyoctanoate (PHO), and other
synthetic polymers), the wild-type S25H strain degraded PHO, as did BK25H-LCC, while S25H-LCC degraded
PLA. Building on these growth observations, we further examined the ability of LCC-expressing Pseudomonas
strains to degrade plastic polymers through biofilm formation. Scanning electron microscopy revealed
substantial surface degradation, particularly on PET (BR5H-LCC biofilms) and PLA/LDPE (BK25H-LCC
biofilms), with visible textural alterations and increased susceptibility to heat-induced damage.
These findings highlight the potential of Pseudomonas biofilm forming strains expressing LCC as a promising
biotechnological strategy for the degradation or surface modification of plastic materials.BOOK OF ABSTRACTS: 17th International Symposium on
Biocatalysis and Biotransformations
Basel, Switzerland, June 29-July 3, 202
Live RNA imaging and knockdown using programmable CRISPR-Cas13 (LiveRNA)
No full textNon-coding transcripts of protein-coding genes represent an underexplored area in transcriptomics. LiveRNA project will utilize expertise of our partner in CRISPR/Cas13 technology for live RNA imaging and knockdown of ANXA3-204 transcript with retained intron, as proof-ofprinciple. By visualizing and altering its expression, we will study its functions in colorectal cancer. Staff mobility will increase methodological capacities of the Serbian team and strengthen collaboration with diaspora.Principal Investigator: Dr. Jovana Despotović, IMGGEParticipants from IMGGE: Dunja Pavlović i Anastasija BubanjaDuration period: 2025 - 202
Gut microbiota and autism spectrum disorder: bioinformatic meta-analysis
No full textAutism spectrum disorder, a neurodevelopmental disorder that manifests from early childhood, is now diagnosed in one in every 100 children worldwide, with exact causes still remaining unclear. Certain scientists associate it with disruptions in the gut microbiota (dysbiosis). However, as different studies report varying results, there is still no clear consensus on which microbial markers of dysbiosis are linked to autism spectrum disorder. The use of different laboratory techniques and subsequent bioinformatic analysis affect the results. Therefore, the aim of this study was to obtain clearer understanding of potential microbial markers of autism spectrum disorder by applying uniform bioinformatic protocol on integrated datasets. After filtering the available literature, a bioinformatic meta-analysis was conducted combining raw data obtained from two independent studies. These studies compared gut bacterial communities of children with autism spectrum disorder and healthy children, by analysing the V3-V4 region of the 16S rRNA gene. The raw sequences from the 188 samples (96 from one study and 92 from the other) were processed using the QIIME2 platform, resulting in amplicon sequence variants (ASVs). Alpha and beta diversity metrics were determined, and taxonomic identification was performed by comparing ASVs with sequences in the Silva reference database. The meta-analysis results revealed significant differences in beta diversity between the groups, as well as in certain bacterial genera, many of which remain uncultured and poorly described. Harmonizing approaches to microbiota analysis is crucial for the reliable identification of dysbiosis markers, which could contribute to novel preventive and therapeutic strategies for autism spectrum disorder.BeCELS 2025: Belgrade Conference for Early-Career Life Scientists, taking place on Friday, September 5, 2025, at the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in Belgrad
Structural characteristics and terminal galactose expression of monoclonal IgA paraproteins from sera of multiple myeloma patients
No full textIgA myeloma accounts for approximately 20% of all myeloma cases and is generally associated with a less favorable prognosis than other subtypes. Some complications specific to IgA myeloma might be linked to the structural features of monoclonal IgA. These include a high degree of polymerization, which can cause hyperviscosity syndrome, and reduced expression of galactose (Gal) residues on the glycans of the IgA hinge region, leading to IgA deposition and nephropathy. To assess the level of polymerization and galactose expression, we isolated IgA from the sera of 15 myeloma patients using affinity chromatography on Protein M agarose. Non-reducing SDSPAGE revealed two or more fractions, with molecular weights of 160 kDa and higher. Western blot analysis confirmed the presence of IgA monomers and dimers in all samples. Galactose expression was evaluated by lectin blotting using Ricinus communis agglutinin I (RCA I), a Dgalactose-binding lectin. RCA I blotting demonstrated that both monomeric and polymeric IgA forms expressed galactose, although the intensity notably varied among samples. Our results suggest that the structural properties of myeloma IgA are heterogeneous. To determine whether these differences influence disease progression, we plan to correlate IgA structural features with patients’ clinical data. Further, in-depth glycoprotein analyses will be carried out using mass spectrometry in collaboration with the Horizon CanSERVBeCELS 2025: Belgrade Conference for Early-Career Life Scientists, taking place on Friday, September 5, 2025, at the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in Belgrad
Transcriptomes and proteomes of the desiccation-tolerant resurrection plant Ramonda serbica
No full textAn extreme loss of cellular water, desiccation (5-10% of relative water content), leads to
denaturation, aggregation and degradation of proteins and affects the fluidity and integrity of
cellular membranes. Plant species that can survive these extreme conditions and fully resume
metabolic functions upon rewatering in a very short period, even within 48 h, are called
resurrection plants. Ramonda serbica Panč., an ancient and endemic species, belongs to
resurrection plants.
The goal of this study was to identify key candidates and pathways involved in desiccation
tolerance in R. serbica, using differential transcriptomic and proteomic approaches. Based on de
novo transcriptome analysis, 68694 differentially expressed genes were detected upon desiccation.
Shotgun proteomics revealed 1192 different protein groups; among them, 408 showed statistically
significant differences in abundance between hydrated (HL) and desiccated leaves (DL). The
majority of proteins related to photosynthetic processes were less abundant in the leaves upon
desiccation. The amounts of H2O2 scavenging enzymes, including ascorbate-glutathione cycle
components, catalases, peroxiredoxins, Fe-, and Mn- superoxide dismutase (SOD) were reduced
in DL. However, four Cu/ZnSOD isoforms, three polyphenol oxidases, six germin-like proteins,
and 22 late embryogenesis abundant proteins were desiccation-inducible. Our study demonstrated
that desiccation tolerance in R. serbica is a complex, species-specific process orchestrated by
several metabolic pathways and regulatory networks. These findings can have a significant impact
on improving the drought tolerance of crops grown in arid regions.BeCELS 2025: Belgrade Conference for Early-Career Life Scientists, taking place on Friday, September 5, 2025, at the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in Belgrad