Design and biofabrication of bacterial living materials with robust and multiplexed biosensing capabilities

Usai, Francesca; Loi, Giada; Scocozza, Franca; Bellato, Massimo; Castagliuolo, Ignazio; Conti, Michele; Pasotti, Lorenzo

doi:10.1016/j.mtbio.2022.100526

Cited by 8 publications

(4 citation statements)

References 47 publications

(53 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PB5741 contains an IPTG-inducible P hyspank promoter [59] upstream of the pgs operon genes, enabling user-defined tuning of operon expression and removing all the existing regulations. PB5741 has a strong IPTG-inducible mucoid phenotype, qualitatively indicating γ-PGA production [60]. This behavior motivated us to quantitatively characterize γ-PGA production in this strain under different conditions.…”

Section: Engineering the γ-Pga Biosynthetic Operon Regulation Using A...mentioning

confidence: 91%

See 1 more Smart Citation

Metabolic Engineering of Bacillus subtilis for the Production of Poly-γ-Glutamic Acid from Glycerol Feedstock

Pasotti,

Massaiu,

Magni

et al. 2024

Fermentation

Self Cite

View full text Add to dashboard Cite

Poly-γ-glutamic acid (γ-PGA) is an attractive biopolymer for medical, agri-food, and environmental applications. Although microbial synthesis by Bacilli fed on waste streams has been widely adopted, the obtainment of efficient sustainable production processes is still under investigation by bioprocess and metabolic engineering approaches. The abundant glycerol-rich waste generated in the biodiesel industry can be used as a carbon source for γ-PGA production. Here, we studied fermentation performance in different engineered Bacillus subtilis strains in glycerol-based media, considering a swrA+ degU32Hy mutant as the initial producer strain and glucose-based media for comparison. Modifications included engineering the biosynthetic pgs operon regulation (replacing its native promoter with Physpank), precursor accumulation (sucCD or odhAB deletion), and enhanced glutamate racemization (racE overexpression), predicted as crucial reactions by genome-scale model simulations. All interventions increased productivity in glucose-based media, with Physpank-pgs ∆sucCD showing the highest γ-PGA titer (52 g/L). Weaker effects were observed in glycerol-based media: ∆sucCD and Physpank-pgs led to slight improvements under low- and high-glutamate conditions, respectively, reaching ~22 g/L γ-PGA (26% increase). No performance decrease was detected by replacing pure glycerol with crude glycerol waste from a biodiesel plant, and by a 30-fold scale-up. These results may be relevant for improving industrial γ-PGA production efficiency and process sustainability using waste feedstock. The performance differences observed between glucose and glycerol media also motivate additional computational and experimental studies to design metabolically optimized strains.

show abstract

Section: Engineering the γ-Pga Biosynthetic Operon Regulation Using A...mentioning

confidence: 91%

“…Fermentation 2024, 10, x FOR PEER REVIEW 9 of 1 existing regulations. PB5741 has a strong IPTG-inducible mucoid phenotype, qualitativel indicating γ-PGA production [60]. This behavior motivated us to quantitatively charac terize γ-PGA production in this strain under different conditions.…”

Section: γ-Pga Production In Glucose-based Mediamentioning

confidence: 95%

Metabolic Engineering of Bacillus subtilis for the Production of Poly-γ-Glutamic Acid from Glycerol Feedstock

Pasotti,

Massaiu,

Magni

et al. 2024

Fermentation

Self Cite

View full text Add to dashboard Cite

show abstract

“…Another successful development of ELM has been illustrated in the prototype of hydrogel encapsulated engineered multiple bacterial strains of E. coli and B. subtilis by 3D bioprinting. The function of the hydrogel‐encapsulated ELM demonstrated detectable bacteria in the environmental sample (soil and water) and in the clinical sample (bronchial aspirate sample with P. aeruginosa infection), demonstrating red color pigment due to chemical induction 139 …”

Section: Application Of Polymeric Film and Hydrogelmentioning

confidence: 99%

Impact of polymeric films and hydrogels: Physical characteristics on bacterial growth

Techakasikornpanich,

Jangpatarapongsa,

Polpanich

et al. 2024

Polymers for Advanced Techs

View full text Add to dashboard Cite

Hydrogels find diverse applications in manipulating bacteria, and serving purposes like elevation, maintenance, and elimination. Several factors of hydrogel have been studied in the benefits of antibacterial activity. Factors such as hydrogel stiffness and roughness gain significance in surface coating, influencing bacterial behavior. However, the intricate interplay of hydrogel stiffness, roughness, polymer types, and bacterial species necessitates further exploration. The choice of polymer is dictated by the specific objectives, particularly in antibacterial scenarios where polymers with positive charge, hydrophilicity, and acidity prove effective. These properties induce robust electrostatic and hydrophobic/hydrophilic interactions, along with pH‐induced cell membrane damage, collectively contributing to hindered bacterial adhesion and growth. Additionally, extracellular polymeric substances (EPS) emerge as pivotal influencers in bacterial adhesion and proliferation. EPS production alters bacterial surfaces, fostering connections between bacteria and facilitating biofilm formation. The hydrophobic nature of EPS further complicates bacterial interactions with surface materials, emphasizing the nuanced interplay of hydrophilic and hydrophobic forces in bacterial adhesion. Herein, this work article has reviewed the related study of each physical property related to antibacterial property on the surface of the hydrogel. Moreover, this work also illustrates applications of the antibacterial properties of hydrogel for medical and surface treatment, including wound healing, food packaging, and surface coating. Additionally, the bacteria growing on hydrogel for engineered living materials, have been updated in various applications.

show abstract

“…More recently, 3D bioprinting has also been used to produce functional materials in which microorganisms are cultured. The advancement and integration of bioprinting techniques specifically for the printing of microorganisms offers the potential for a completely new generation of biologically produced functional materials [ 7 – 9 ]. Utilizing and further developing 3D bioprinting to create functional bacteria-laden structures can help to solve various challenges in diverse application fields, such as therapeutic devices, environmental engineering, and industrial biomanufacturing [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

3D bioprinting of microorganisms: principles and applications

Herzog,

Franke,

Lai

et al. 2024

Bioprocess Biosyst Eng

View full text Add to dashboard Cite

In recent years, the ability to create intricate, live tissues and organs has been made possible thanks to three-dimensional (3D) bioprinting. Although tissue engineering has received a lot of attention, there is growing interest in the use of 3D bioprinting for microorganisms. Microorganisms like bacteria, fungi, and algae, are essential to many industrial bioprocesses, such as bioremediation as well as the manufacture of chemicals, biomaterials, and pharmaceuticals. This review covers current developments in 3D bioprinting methods for microorganisms. We go over the bioink compositions designed to promote microbial viability and growth, taking into account factors like nutrient delivery, oxygen supply, and waste elimination. Additionally, we investigate the most important bioprinting techniques, including extrusion-based, inkjet, and laser-assisted approaches, as well as their suitability with various kinds of microorganisms. We also investigate the possible applications of 3D bioprinted microbes. These range from constructing synthetic microbial consortia for improved metabolic pathway combinations to designing spatially patterned microbial communities for enhanced bioremediation and bioprocessing. We also look at the potential for 3D bioprinting to advance microbial research, including the creation of defined microenvironments to observe microbial behavior. In conclusion, the 3D bioprinting of microorganisms marks a paradigm leap in microbial bioprocess engineering and has the potential to transform many application areas. The ability to design the spatial arrangement of various microorganisms in functional structures offers unprecedented possibilities and ultimately will drive innovation.

show abstract

Design and biofabrication of bacterial living materials with robust and multiplexed biosensing capabilities

Cited by 8 publications

References 47 publications

Metabolic Engineering of Bacillus subtilis for the Production of Poly-γ-Glutamic Acid from Glycerol Feedstock

Metabolic Engineering of Bacillus subtilis for the Production of Poly-γ-Glutamic Acid from Glycerol Feedstock

Impact of polymeric films and hydrogels: Physical characteristics on bacterial growth

3D bioprinting of microorganisms: principles and applications

Contact Info

Product

Resources

About