Biology subjects

Simoens, K.

Publications and source records attributed to Simoens, K..

6 recordsLinked to original sources

bioRxiv2025

Phage-mediated just-in-time circuit amplification and delivery for recombinant expression of toxic proteins

In order to address outstanding challenges in bacterial bioreactor bioprocessing, we have developed an approach that alleviates the need for chemical inducers and selective agents (such as antibiotics), and mitigates leaky expression of the production circuit. More specifically, temperate bacteriophage Lambda ({lambda}) was engineered as a chassis to just-in-time amplify and deliver a production circuit to a wild-type Escherichia coli population. Since the expression of the production pathway is engineered to be dependent on {lambda}s central regulatory switch towards the lysogenic state, {lambda} will first lytically amplify in the background without significantly altering overall bioreactor growth dynamics or gene expression. Only when {lambda} eventually starts outnumbering the host population, its natural switch to lysogeny will actively install and trigger the production circuit. As such, the {lambda} chassis serves as an intrinsic self-amplifying inducer that meanwhile rules out any leaky expression. The impact of the initial phage-to-host ratio on population dynamics and production efficiency was assessed experimentally and through ODE modeling, revealing that it can be used to fine-tune the trade-off between gene dosage and biomass conversion into virions. We demonstrate that this leakproof approach is particularly promising for the expression of toxic proteins such as Benzonase, a highly potent DNase and RNase.

synthetic biology↗

bioRxiv2025

BIO-SPEC: an open-source bench-top parallel bioreactor system.

The BIO-SPEC is an open-source, cost-effective, and modular bench-top bioreactor system designed for batch, sequencing batch, and chemostat cultivation. Featuring thermoelectric condensers to eliminate the need for a chiller, it ensures stable long-term operation. Controlled by a Raspberry Pi, the BIO-SPEC offers flexibility in headplate design, gas supply, and feeding strategies, making it a versatile alternative to high-cost commercial systems. This paper details the design, construction, and validation of the BIO-SPEC system, demonstrating its potential to advance microbiology and bioprocessing research through accessible and reliable hardware at a fraction of the cost.

microbiology↗

bioRxiv2024

Discovery of alternative stable states in a synthetic human gut microbial community

AbstractSeveral human-associated microbial communities exist in multiple configurations and can change their composition in response to perturbations, remaining in an altered state even after the perturbation ends. Multistability has been previously proposed to explain this behavior for gut microbiota in particular, but has not been clearly demonstrated experimentally. Here, we first investigated the life history strategies of three common human gut bacteria to identify mechanisms driving alternative states. We then used this data to build and parameterize a kinetic model, which predicted that alternative states emerge due to phenotype switching between subpopulations of the same species. Perturbation experiments supported these predictions, and confirmed the existence of alternative states. Finally, simulations showed that phenotype switching can also explain alternative states in larger communities. Thus, a transient perturbation combined with metabolic flexibility is sufficient for alternative communities to emerge, implying that they are not necessarily explained by differences between individuals. One-Sentence SummaryWe demonstrate the existence of alternative states in a human gut microbial community and propose phenotype switching as a mechanism explaining their emergence.

microbiology↗

bioRxiv2024

Establishing a co-culture aggregate of N-cycle bacteria to elucidate flocculation in biological wastewater treatment

Biological flocculation is a complex phenomenon that is often treated as a black box. As a result, flocculation problems are usually remediated without knowledge of the exact causes. We show that it is feasible to exploit a model (N-cycle) consortium with reduced complexity to fundamentally study bioflocculation. Strong nitrifier microcolonies were formed during oxic/anoxic cycles in sequencing batch reactors, using alginate entrapment as a cell retention system. After release of these aggregates into suspension, macroclusters with flocs of the denitrifier were observed. These results suggest that a living model of a full-scale activated sludge floc can be built through the use of this bottom-up approach. By eliminating shifts in the microbial community, the applied experimental conditions have a more direct effect on the observations.

microbiology↗

bioRxiv2023

Enhanced protein secretion in reduced genome strains of Streptomyces lividans

S. lividans TK24 is a popular host for the production of small molecules and for the secretion of heterologous proteins. TK24 has a large genome with at least 29 secondary metabolite gene clusters that are non-essential for viability and undergo complex regulation. To optimize heterologous protein secretion, we previously constructed ten chassis strains that are devoid of several secondary metabolite gene clusters. Genome reduction was aimed at reducing carbon flow to secondary metabolites and pigmentation in the spent growth medium and improving colony morphology. Strains RG1.0-RG1.10 contain various deletion combinations of the blue actinorhodin cluster (act), the calcium-dependent antibiotic (cda), the undecylprodigiosin (red) and coelimycin A (cpk) clusters, the melanin cluster (mel), the matAB genes that affect mycelial aggregation and the non-essential sigma factor hrdD that controls the transcription of Act and Red regulatory proteins. Two derivative strains, RG1.5 and 1.9, showed a [~]15% reduction in growth rate, >2-fold increase in the total mass yield of their native secretome and altered abundance of several specific proteins compared with TK24. Metabolomics and RNAseq analysis revealed that genome reduction led to rapid cessation of growth due to aminoacid depletion and caused both redox and cell envelope stresses, upregulation of the Sec-pathway components secDF and chaperones and a cell envelope two component regulator. RG1.9 maintained elevated heterologous secretion of mRFP and mTNF by 12-70%. An integrated model is presented linking genome reduction and enhanced secretion.

systems biology↗

bioRxiv2023

Starvation response strategies of human gut bacteria Bacteroides thetaiotaomicron and Roseburia intestinalis and their impact on co-culture dynamics

Bacterial growth often alters the environment, which in turn can impact interspecies interactions among bacteria. Here, we used an in vitro batch system containing mucin beads to emulate the dynamic host environment and to study its impact on the interactions between two abundant and prevalent human gut bacteria, the primary fermenter Bacteroides thetaiotaomicron and the butyrate producer Roseburia intestinalis. By combining machine learning and flow cytometry, we found that the number of viable B. thetaiotaomicron cells decreases with glucose consumption due to acid production, while R. intestinalis survives post-glucose depletion by entering a slow growth mode. Both species attach to mucin beads, but only viable cell counts of B. thetaiotaomicron increase significantly. The number of viable co-culture cells varies significantly over time compared to those of monocultures. A combination of targeted metabolomics and RNA-seq showed that the slow growth mode of R. intestinalis represents a diauxic shift towards acetate and lactate consumption, whereas B. thetaiotaomicron survives glucose depletion and low pH by foraging on mucin sugars. In addition, most of the mucin monosaccharides we tested inhibited the growth of R. intestinalis but not B. thetaiotaomicron. We encoded these causal relationships in a kinetic model, which reproduced the observed dynamics. In summary, we explored how R. intestinalis and B. thetaiotaomicron respond to nutrient scarcity and how this affects their dynamics. We highlight the importance of understanding bacterial metabolic strategies to effectively modulate microbial dynamics in changing conditions.

microbiology↗