Biology subjects

Solomon, K. V.

Publications and source records attributed to Solomon, K. V..

4 recordsLinked to original sources

bioRxiv2026

Programmable assembly of higher-order DNA nanostructures from microbial ssDNA staple libraries

Bottom-up manufacturing of structural DNA nanotechnology requires a long single-stranded DNA (ssDNA) scaffold and hundreds of short ([~]30 nt) ssDNA staples. However, scaling production remains bottlenecked by the high economic cost and environmental footprint of solid-phase chemical staple synthesis. To address these limitations, a phage-free, biological nanomanufacturing platform engineered in Escherichia coli is developed here. Two intracellular strategies for producing programmable ssDNA were systematically evaluated: retron-based multicopy ssDNA (msDNA) synthesis via the Ec67 system and plasmid-encoded rolling circle replication (RCR). While native structural topology constraints within the retron (msd) cassette limit its sequence-design flexibility, the alternative RCR-driven engine successfully decouples ssDNA replication from sequence secondary structures, enabling the synthesis of arbitrary staples. This RCR platform reliably generates long circular ssDNA (cssDNA) precursors of at least 1.8 kb with exceptional sequence fidelity (>99%). Integrating programmable BseGI cleavage sites allows targeted strand-selective enzymatic processing to cleanly release stoichiometric, origami-grade pools of 32-nt staple strands. Atomic force microscopy (AFM) confirms that these biologically produced staples successfully drive the high-fidelity self-assembly of complex DNA tiles and hollow tubules. Crucially, robust structural folding is demonstrated directly within crude, unpurified cellular lysates, establishing a green, cost-effective framework for the one-pot fabrication of advanced DNA-based nanomaterials.

synthetic biology↗

bioRxiv2025

A high throughput assay to detect enzymatic polyethylene oxidation

Biological plastics deconstruction and upcycling have emerged as a sustainable alternative to traditional recycling technologies for plastics waste. The discovery and engineering of efficient thermostable poly(ethylene terephthalate) (PET) hydrolases has made biological PET recycling possible at scale; however, enzymes for non-PET plastics, which account for approximately 70% of all plastics produced, remain largely undiscovered. To accelerate the discovery of such enzymes, a high throughput screening (HTS) platform is needed. Here, we develop a HTS liquid-based assay to detect one of the first committed steps of polyolefin degradation, oxidation of the C-H bond to an aldehyde. We test 4-hydrazino-7-nitro-2,1,3-benxoxadiozole hydrazine (NBD-H), which reacts with generated aldehydes to form a fluorescent hydrazone, on oxidized low-density polyethylene (LDPE) films. Hydrazone generation correlated well with established carbonyl index metrics for polymer oxidation (R2 = 0.97). Moreover, we demonstrate that the probe reliably identifies LDPE-active dye decolorizing peroxidases (DyPs) that generate aldehydes on LDPE films, serving as effective screen as demonstrated by a receiver operating characteristic area under the curve of 0.95. Due to the rapid fluorescent readout and parallelization in microarray plates, this assay enables screening thousands of enzymes in 24 hours compared to time-consuming established approaches, accelerating discovery of enzymes that catalyze the first step of polyolefin biodeconstruction.

biochemistry↗

bioRxiv2025

Biological polyethylene deconstruction initiated by oxidation from DyP peroxidases

Polyethylene (PE) is the most commonly used plastic on Earth due to its favorable material properties such as high ductility, mechanical strength, and bond homogeneity that make the material resistant to deconstruction. However, the lack of robust recycling infrastructure for PE end-of-life management is leading to an estimated 4 million tons of environmental accumulation annually, with implications for human and environmental health. Biological deconstruction and upcycling could potentially aid in PE waste management by allowing for high-yield conversion of waste plastics to high value products, although such processes are not yet possible. In this work, we mined the gut of low-density PE (LDPE) fed mealworms that can reduce LDPE molecular weight by >40% and discovered dye decolorizing peroxidases (DyPs) that oxidized LDPE, initiating biological deconstruction. A plastic-active DyP is characterized by a hydrophobic loop near its active site that helps mediate binding and tunes activity. LDPE oxidation is driven by surface exposed residues proximal to the active site enabling activity on polymeric substrates. Our work provides robust evidence for enzymatic LDPE deconstruction and identifies molecular targets for further development to realize scalable biological LDPE upcycling. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=41 SRC="FIGDIR/small/640435v1_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@146b8fborg.highwire.dtl.DTLVardef@1a72adorg.highwire.dtl.DTLVardef@5ece5forg.highwire.dtl.DTLVardef@3ee20a_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2024

Hydrophobins from Aspergillus mediate fungal interactions with microplastics

Microplastics present myriad ecological and human health risks including serving as a vector for pathogens in human and animal food chains. However, the specific mechanisms by which pathogenic fungi colonize these microplastics have yet to be explored. In this work, we examine the opportunistic fungal pathogen, Aspergillus fumigatus, and other common soil and marine Aspergilli, which we found bind microplastics tightly. Up to 3.85+/-1.48 g microplastic plastic/g fungi were bound and flocculated for polypropylene (PP), polyethylene (PE), and polyethylene terephthalate (PET) powders and particles ranging in size from 0.05 - 5 mm. Gene knockouts revealed hydrophobins as a key biomolecule driving microplastic-fungi binding. Moreover, purified hydrophobins were still able to flocculate microplastics independent of the fungus. Our work elucidates a role for hydrophobins in fungal colonization of microplastics and highlights a potential target for mitigating the harm of microplastics through engineered fungal-microplastic interactions. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/622132v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@13eb679org.highwire.dtl.DTLVardef@3dc9c0org.highwire.dtl.DTLVardef@888c96org.highwire.dtl.DTLVardef@c5e1aa_HPS_FORMAT_FIGEXP M_FIG C_FIG Significance StatementMicroplastics pose serious ecological and human health effects by introducing pathogens and toxins into animal and human food chains. Many pathogenic microorganisms preferentially form biofilms on microplastic particles that are then ingested. Here, we demonstrate that hydrophobins, highly hydrophobic, cell surface proteins, enable microplastic binding and colonization by the opportunistic pathogen Aspergillus fumigatus and other fungi within the Aspergillus genus. Our work recognizes a novel role for hydrophobin proteins, identifying potential strategies for pathogen control and protein-based microplastics recovery.

molecular biology↗