Nonsterile Process for Biohydrogen Production: Recent Updates, Challenges, and Opportunities.

Hydrogen (H2), a clean and versatile energy carrier, has recently gained significant attention as a potential solution for reducing carbon emissions and promoting sustainable energy systems. The yield and efficiency of the biological H2 production process primarily depend on sterilization conditions. Various strategies, such as heat inactivation and membrane-based sterilization, have been used to achieve desirable yields via microbial fermentation. Almost every failed biotransformation process is linked to nonsterile conditions at any reaction stage. Therefore, the production of renewable biofuels as alternatives to fossil fuels is more attractive. Pure sugars have been widely documented as a costly feedstock for H2 production under sterile conditions. Biotransformation under nonsterile conditions is more desirable for stable and sustainable operation. Low-cost feeds, such as biowaste, are considered suitable alternatives, but they require appropriate sterilization to overcome the limitations of inherited or contaminating microbes during H2 production. This article describes the status of microbial fermentative processes for H2 production under nonsterile conditions and discusses strategies to improve such processes for sustainable, cleaner production.

Trametes versicolor Laccase-Based Magnetic Inorganic-Protein Hybrid Nanobiocatalyst for Efficient Decolorization of Dyes in the Presence of Inhibitors.

In the present investigation, an ecofriendly magnetic inorganic-protein hybrid system-based enzyme immobilization was developed using partially purified laccase from Trametes versicolor (TvLac), Fe3O4 nanoparticles, and manganese (Mn), and was successfully applied for synthetic dye decolorization in the presence of enzyme inhibitors. After the partial purification of crude TvLac, the specific enzyme activity reached 212 U∙mg total protein-1. The synthesized Fe3O4/Mn3(PO4)2-laccase (Fe3O4/Mn-TvLac) and Mn3(PO4)2-laccase (Mn-TvLac) nanoflowers (NFs) exhibited encapsulation yields of 85.5% and 90.3%, respectively, with relative activities of 245% and 260%, respectively, compared with those of free TvLac. One-pot synthesized Fe3O4/Mn-TvLac exhibited significant improvements in catalytic properties and stability compared to those of the free enzyme. Fe3O4/Mn-TvLac retained a significantly higher residual activity of 96.8% over that of Mn-TvLac (47.1%) after 10 reuse cycles. The NFs showed potential for the efficient decolorization of synthetic dyes in the presence of enzyme inhibitors. For up to five reuse cycles, Fe3O4/Mn-TvLac retained a decolorization potential of 81.1% and 86.3% for Coomassie Brilliant Blue R-250 and xylene cyanol, respectively. The synthesized Fe3O4/Mn-TvLac showed a lower acute toxicity towards Vibrio fischeri than pure Fe3O4 nanoparticles did. This is the first report of the one-pot synthesis of biofriendly magnetic protein-inorganic hybrids using partially purified TvLac and Mn.

Antimicrobial photodynamic therapy on Staphylococcus aureus and Escherichia coli using malachite green encapsulated mesoporous silica nanoparticles: an in vitro study.

BACKGROUND: Rise in the number of healthcare associated or hospital acquired infections is a major problem affecting the global healthcare sector. We evaluated superior antibacterial and antibiofilm photodynamic therapy (aPDT) using malachite green encapsulated mesoporous silica nanoparticles (MG-MSN) against Staphylococcus aureus and Escherichia coli, which are known to be major causative agents of nosocomial infections. METHODS: Malachite green (MG) was encapsulated on mesoporous silica nanoparticles (MSN). Fourier-transform infrared spectroscopy, Transmission electron microscopy, and spectroscopic analysis were performed to characterize the MG-MSN. The antimicrobial efficacies of MSN, MG, and MG-MSN were investigated and the results were recorded. RESULTS: MG-MSN was effective against both the tested bacteria. S. aureus was more phototoxic to MG-MSN compared to E. coli. The antibiofilm efficacy of MG-MSN on E. coli and S. aureus was also studied. Biofilm inhibition was 65.68 ± 2.62% in E. coli and 79.66 ± 3.82% in S. aureus. Cell viability assay, exopolysaccharides quantification, and confocal laser scanning microscopy studies also revealed the enhanced antibiofilm activity of MG-MSN when used as a potential photosensitizer for aPDT. This study can be extended to eradicate these strains from localized superficial infections and medical appliances, preventing nosocomial infections.

Malachite green-conjugated multi-walled carbon nanotubes potentiate antimicrobial photodynamic inactivation of planktonic cells and biofilms of Pseudomonas aeruginosa and Staphylococcus aureus.

Purpose: Infections associated with medical devices that are caused by biofilms remain a considerable challenge for health care systems owing to their multidrug resistance patterns. Biofilms of Pseudomonas aeruginosa and Staphylococcus aureus can result in life-threatening situations which are tough to eliminate by traditional methods. Antimicrobial photodynamic inactivation (aPDT) constitutes an alternative method of killing deadly pathogens and their biofilms using reactive oxygen species (ROS). This study investigated the efficacy of enhanced in vitro aPDT of P. aeruginosa and S. aureus using malachite green conjugated to carboxyl-functionalized multi-walled carbon nanotubes (MGCNT). Both the planktonic cells and biofilms of test bacteria were demonstrated to be susceptible to the MGCNT conjugate. These MGCNT conjugates may thus be employed as a facile strategy for designing antibacterial and anti-biofilm coatings to prevent the infections associated with medical devices. Methods: Conjugation of the cationic dye malachite green to carbon nanotube was studied by UV-visible spectroscopy, high-resolution transmission electron microscopy, and Fourier transform infrared spectrometry. P. aeruginosa and S. aureus photodestruction were studied using MGCNT conjugate irradiated for 3 mins with a red laser of wavelength 660 nm and radiant exposure of 58.49 J cm-2. Results: Upon MGCNT treatment, S. aureus and P. aeruginosa were reduced by 5.16 and 5.55 log10 , respectively. Compared to free dye, treatment with MGCNT afforded improved phototoxicity against test bacteria, concomitant with greater ROS production. The results revealed improved biofilm inhibition, exopolysaccharide inhibition, and reduced cell viability in test bacteria treated with MGCNT conjugate. P. aeruginosa and S. aureus biofilms were considerably reduced to 60.20±2.48% and 67.59±3.53%, respectively. Enhanced relative MGCNT phototoxicity in test bacteria was confirmed using confocal laser scanning microscopy. Conclusion: The findings indicated that MGCNT conjugate could be useful to eliminate the biofilms formed on medical devices by S. aureus and P. aeruginosa.

Antimicrobial photodynamic activity of toluidine blue-carbon nanotube conjugate against Pseudomonas aeruginosa and Staphylococcus aureus - Understanding the mechanism of action.

BACKGROUND: The emergence of drug-resistant bacterial strains has raised the need to develop alternative treatment modalities to combat infectious diseases. Antimicrobial photodynamic therapy (aPDT) is an alternative to conventional treatment modalities. aPDT integrates a photosensitizer, which, after exposure to light of an appropriate wavelength, leads to the generation of cytotoxic reactive oxygen species (ROS). METHODS: The aim of the present study was to synthesize a toluidine blue/multiwalled carbon nanotube conjugate (TBCNT) for enhanced photoinactivation of Pseudomonas aeruginosa and Staphylococcus aureus. Synthesized TBCNT conjugate was characterized and its antibacterial and antibiofilm activity was determined. RESULTS: During TBCNT synthesis, dye loading, and entrapment efficiency of the CNT were 12.04 ±â€¯0.55% and 48.99 ±â€¯2.33%, respectively. The photo-destruction of planktonic cells of the test bacteria was performed by exposure to a 125 mW red laser with a wavelength of 670 nm (radiant exposure of 58.49 J/cm2) for 3 min. Photoinactivation using TBCNT resulted in a 4.91- and 5.47-log10 reduction in P. aeruginosa and S. aureus, respectively. The mechanism of this aPDT was studied by measuring intracellular ROS generation, protein leakage, and lipid peroxidation in the test bacteria after light irradiation. The antibiofilm activity of TBCNT after light exposure was 69.94% and 75.54% for P. aeruginosa and S. aureus, respectively. Photoinactivation of test bacteria treated with TBCNT reduced cell viability and exopolysaccharide production. Confocal laser-scanning microscopy revealed a significant biofilm inhibition efficacy of the TBCNT conjugate. CONCLUSION: Therefore, TBCNT conjugates may be used for the eradication of P. aeruginosa and S. aureus biofilms.

Antioxidant, anti-quorum sensing and anti-biofilm potential of ethanolic leaf extract of Phrynium capitatum and Dryptes indica

Objective: To investigate the antioxidant and anti-infective potential of Phrynium capitatum and Dryptes indica extract. Methods: The antioxidant potentials were determined by DPPH radical scavenging, reducing power, hydroxyl radical scavenging and total antioxidant assays. We further examined anti-quorum sensing activity and inhibition of synthesis of pathogenic factor of Chromobacterium violaceum and Pseudomonas aeruginosa PAO1. Bioactive compounds were determined using gas chromatography–mass spectrometry analysis. In silico analysis was conducted to determine the binding affinity of bioactive compounds of plant extracts for the quorum sensing regulatory receptor LasR. Results: DPPH assay showed that the ethanolic extract of Phrynium capitatum and Dryptes indica at 500 μg/mL showed (86.96 ± 4.07)％ and (74.83 ± 3.47)％ inhibition, respectively. Hydroxyl radical scavenging assay showed (73.17 ± 3.03)％ and (62.63 ± 4.59)％ activity, respectively. The ethanolic extract of Phrynium capitatum and Dryptes indica showed high level of attenuation of quorum sensing regulated pyocyanin production. Confocal laser scanning microscopic analysis revealed that the extracts had the potential to effectively inhibit biofilm formation of Pseudomonas aeruginosa. Molecular docking analysis showed a better binding affinity of bioactive compounds from the extracts for the structure of LasR protein of Pseudomonas aeruginosa. Conclusions: The ethanolic extracts of Phrynium capitatum and Dryptes indica possess antioxidant activity and the potential to inhibit the quorum sensing system and its regulatory virulence traits in Pseudomonas aeruginosa PAO1.

Antimicrobial photodynamic activity of rose bengal conjugated multi walled carbon nanotubes against planktonic cells and biofilm of Escherichia coli.

BACKGROUND: The global threat of antimicrobial resistance especially due to the bacterial biofilms has engaged researchers in the search of new treatment modalities. Antimicrobial photodynamic inactivation (aPDI) is one of the alternative treatment modalities which kills bacteria by generating endogenous reactive oxygen species (ROS). In this work authors evaluated the antibacterial and antibiofilm efficacy of rose Bengal (RB) conjugated to CNT against E. coli. METHODS: The interaction of anionic dye to the CNT was studied using UV-vis spectroscopy, HRTEM, FTIR and spectrofluorometry. Phototoxicity of RBCNT conjugate against E. coli was studied using a green light of 50 mW and radiant exposure of 1674.7 J/ cm2 for 10 min. The antibiofilm activity and mechanism of action of RBCNT conjugate in presence of light was evaluated. RESULTS: The loading and encapsulation was found to be 15.46 ± 1.05 and 61.85 ± 4.23% respectively. The photodynamic inactivation of planktonic cells of E. coli was found to 5.46 and 3.56 log10 CFU/ml reduction on treatment with RBCNT and RB respectively. The conjugate also exhibited efficient and enhanced antibiofilm activity against E. coli. The study of mechanism of action has confirmed cell membrane and DNA damage were the two main targets of aPDI. CONCLUSION: This report has concluded the efficient photodynamic inactivation occurred in Gram negative bacteria E. coli due to the increased production of ROS inside the bacterial cells. Hence, the newly synthesized RBCNT conjugate can be efficiently utilized to control infections caused by E. coli.