Potential Biotechnological Applications Research Articles

Screening and evaluation of the ruminal cellulolytic bacteria and their potential application as probiotics.

Rumen microbiologists are looking for new probiotics to improve the digestibility of livestock diets. This study intended to screen and evaluate the ruminal cellulolytic bacteria (CBs) and their potential application as probiotics. Microbial culture and molecular techniques performed to isolate CBs from the rumen of camels, deer and rams. Their antibacterial and antibiogram tests were done using disc diffusion method. Their potential to degrade cellulose, starch, tannin and protein were investigated using clear zone halo, and spectrophotometric techniques. Bilious, saline, and acidic broth media were used to study the resistance of isolates in intestinal conditions. The phylogenetic analysis revealed that the strains belonged to Firmicutes and Proteobacteria phyla, Citrobacter murliniae, Ornithinibacillus bavariensis, C. braakii, and Bacillus subtilis. The highest cellulase (CAS) activity was recorded by C. murliniae Dez wildlife13A (2.98 UmL-1), whereas C. braakii Loot desert 111A (1.14 Uml-1) was produced the lowest enzyme. The isolates were highly resistant to synthetic conditions of intestine (pH 2.5-3.5, bile 0.3-2%), as well as tolerated higher concentrations of NaCl (up to 10%). They effectively inhibited standard pathogen strains, and showed sensitivity to the used antibiotics. This study reports the cellulolytic O. bavariensis Tabbas desert 32A for the first time from the rumen, which will have potential biotechnological applications.

Comparative Biochemical Profiling and Industrial Application Potentials of Dunaliella salina and Spirulina platensis

The diverse biochemical profiles of microalgae, particularly Dunaliella salina and Spirulina platensis, make them essential to biotechnological fields. These microalgae stand out for their high biomass production and capacity to store important materials including proteins, lipids, polysaccharides, and carotenoids in a variety of environmental settings. This work provides a comprehensive evaluation that compares these two popular microalgal species, closely examining important characteristics such organic matter, pigments, carotenoids, carbohydrates, and proteins, along with moisture content, ash content, and ash-free dry weight. The investigation validated the exceptional ability of Dunaliella salina to store protein and β-carotene, indicating its potential use in nutraceutical applications. On the other hand, Spirulina platensis showed increased protein content, which strengthened its standing as a helpful dietary supplement. Spirulina's somewhat higher carbohydrate content suggests that the biofuel industry may find use for it. By identifying each microalga's distinct characteristics in more detail, proximate analysis provided important new information about their potential applications in biotechnology. This thorough characterisation clarifies their inherent benefits notwithstanding the impact of environmental factors and strain selection on their biochemical makeup. By highlighting these qualities, the study advocates for the sustainable use of Dunaliella salina and Spirulina platensis as answers in the rapidly evolving field of biotechnology, and it lays out a path for their advanced usage in a variety of industries, including food, cosmetics, pharmaceuticals, and biofuels.

Pigments profile and antioxidant potential of extremophile cyanobacteria isolated from the Mexican Volcanic Lake Chichonal

Cyanobacteria are ancient and prolific microorganisms for which the current knowledge remains limited. The present work constitutes the first report on the presence of the cyanobacteria Tolypothrix sp. LEGE 221228, Nostoc sp. LEGE 221229, Scytonema sp. LEGE 221230 and Drouetiella sp. LEGE 221231 in the volcanic crater lake in Chichonal (Chiapas-Mexico), and in the characterization of their pigments profile and antioxidant potential, through the application of biorefinery approaches. The identification of cyanobacteria species was achieved through advanced morphology-based methods for cyanobacterial taxonomy. The HPLC-PDA analysis of the cyanobacteria acetone extracts revealed β-carotene, echinenone and chlorophyll-a as the main pigments. Drouetiella sp. LEGE 221231 presented the highest β-carotene concentration (63.67 μg/mL), total carotenoids (87.92 μg/mL) and chlorophylls (20.88 μg/mL), closely followed by Tolypothrix sp. LEGE 221228 (with 51.37, 75.14 and 14.88 μg/mL, respectively), this being the first report on the carotenoid and chlorophylls profile of these strains. Aqueous extracts revealed the highest content of phycoerythrin in Tolypothrix sp. LEGE 221228 (50.61 mg/g) and of phycocyanin in Nostoc sp. LEGE 221229 (57.66 mg/g), standing out in superoxide anion radical scavenging, while the acetonic ones were more effective in scavenging nitric oxide radical. For both, Tolypothrix sp. LEGE 221228 was the most effective, presenting the lowest IC50 (53.75 and 57.68 μg/mL, respectively). Overall, the present study enriches the knowledge on biodiversity of cyanobacteria from underexplored environments, enhancing their potential biotechnological applications as producers of added value secondary metabolites with interest in food, pharmaceutical and cosmetic industries.

Environmental activity-based protein profiling for function-driven enzyme discovery from natural communities

BackgroundMicrobial communities are important drivers of global biogeochemical cycles, xenobiotic detoxification, as well as organic matter decomposition. Their major metabolic role in ecosystem functioning is ensured by a unique set of enzymes, providing a tremendous yet mostly hidden enzymatic potential. Exploring this enzymatic repertoire is therefore not only relevant for a better understanding of how microorganisms function in their natural environment, and thus for ecological research, but further turns microbial communities, in particular from extreme habitats, into a valuable resource for the discovery of novel enzymes with potential applications in biotechnology. Different strategies for their uncovering such as bioprospecting, which relies mainly on metagenomic approaches in combination with sequence-based bioinformatic analyses, have emerged; yet accurate function prediction of their proteomes and deciphering the in vivo activity of an enzyme remains challenging.ResultsHere, we present environmental activity-based protein profiling (eABPP), a multi-omics approach that extends genome-resolved metagenomics with mass spectrometry-based ABPP. This combination allows direct profiling of environmental community samples in their native habitat and the identification of active enzymes based on their function, even without sequence or structural homologies to annotated enzyme families. eABPP thus bridges the gap between environmental genomics, correct function annotation, and in vivo enzyme activity. As a showcase, we report the successful identification of active thermostable serine hydrolases from eABPP of natural microbial communities from two independent hot springs in Kamchatka, Russia.ConclusionsBy reporting enzyme activities within an ecosystem in their native state, we anticipate that eABPP will not only advance current methodological approaches to sequence homology-guided enzyme discovery from environmental ecosystems for subsequent biocatalyst development but also contributes to the ecological investigation of microbial community interactions by dissecting their underlying molecular mechanisms.

Production, Extraction and Partial Characterization of Natural Pigments from Chryseobacterium sp. kr6 Growing on Feather Meal Biomass

Obtaining natural pigments from microorganisms is an alternative with high potential for biotechnological application. The use of agro-industrial wastes as substrate for cultivations enables a reduction of the production cost and may add value to potentially polluting byproducts. In this work, the extraction of pigments produced by the bacterium Chryseobacterium sp. strain kr6 was evaluated, employing feather meal as the sole carbon source for bacterial growth. The maximum production of the yellow pigments was observed for cultivation at 30 °C, during 48 h, with 5 g/L feather meal. The pigment extraction from the bacterial biomass was performed with the aid of physical methods and the testing of different organic solvents. The conditions that provided better extraction were using ultrasound with acetone as the solvent, reaching a yield of 180 μg/g biomass after optimization. The pigment was partially characterized via UV-visible, FTIR and mass spectroscopy and CIELAB color parameters, suggesting the presence of molecules belonging to the flexirubin group (aryl polyenes). The antioxidant capacity of the pigment was confirmed via the scavenging of DPPH radical and thiobarbituric acid reactive substances (TBARS) methodologies. Moreover, the pigment extract showed antimicrobial activity against Staphylococcus aureus and Enterococcus faecalis.

Beyond methane, new frontiers in anaerobic microbial hydrocarbon utilizing pathways.

Alkanes, single carbon methane to long-chain hydrocarbons (e.g. hexadecane and tetradecane), are important carbon sources to anaerobic microbial communities. In anoxic environments, archaea are known to utilize and produce methane via the methyl-coenzyme M reductase enzyme (MCR). Recent explorations of new environments, like deep sea sediments, that have coupled metagenomics and cultivation experiments revealed divergent MCRs, also referred to as alkyl-coenzyme M reductases (ACRs) in archaea, with similar mechanisms as the C1 utilizing canonical MCR mechanism. These ACR enzymes have been shown to activate other alkanes such as ethane, propane and butane for subsequent degradation. The reversibility of canonical MCRs suggests that these non-methane-activating homologues (ACRs) might have similar reversibility, perhaps mediated by undiscovered lineages that produce alkanes under certain conditions. The discovery of these alternative alkane utilization pathways holds significant promise for a breadth of potential biotechnological applications in bioremediation, energy production and climate change mitigation.

Genomic Differences Associated with Resistance and Virulence in Pseudomonas aeruginosa Isolates from Clinical and Environmental Sites.

Pseudomonas aeruginosa is a pathogen that causes healthcare-associated infections (HAIs) worldwide. It is unclear whether P. aeruginosa isolated from the natural environment has the same pathogenicity and antimicrobial resistance potential as clinical strains. In this study, virulence- and resistance-associated genes were compared in 14 genomic sequences of clinical and environmental isolates of P. aeruginosa using the VFDB, PATRIC, and CARD databases. All isolates were found to share 62% of virulence genes related to adhesion, motility, secretion systems, and quorum sensing and 72.9% of resistance genes related to efflux pumps and membrane permeability. Our results indicate that both types of isolates possess conserved genetic information associated with virulence and resistance mechanisms regardless of the source. However, none of the environmental isolates were associated with high-risk clones (HRCs). These clones (ST235 and ST111) were found only in clinical isolates, which have an impact on human medical epidemiology due to their ability to spread and persist, indicating a correlation between the clinical environment and increased virulence. The genomic variation and antibiotic susceptibility of environmental isolates of P. aeruginosa suggest potential biotechnological applications if obtained from sources that are under surveillance and investigation to limit the emergence and spread of antibiotic resistant strains.

The Presence of Arbuscular Mycorrhizal Fungi in the Rhizosphere of Transgenic Rapeseed Overexpressing a Trichoderma Thkel1 Gene Improves Plant Development and Yield

Most of the plants belonging to the family of Brassicaceae are non-hosts for arbuscular mycorrhizal fungi (AMF). These plants are known to produce glucosinolates (GSL), a group of allelopathic compounds, with a role in plant defense. The overexpression of the Thkel1 from Trichoderma harzianum in rapeseed (BnKel) plants, this gene encoding a protein that shares similarities with Brassicaceae plant’s nitrile-specifier and epithiospecifier proteins, modified GSL metabolism, reducing the accumulation of toxic isothiocyanates due to hydrolysis of these secondary metabolites. Here, we have analyzed the effect of AMF application on the GSL profiles and the development and yield of BnKel plants. Our results showed that the reduction of GSL compounds on transgenic plants was not enough to allow the formation of arbuscules and vesicles characteristics of an AMF mycorrhizal association. However, the inoculation of transgenic rapeseed plants expressing Thkel1 with AMF improved seed yield and fatty acid composition of the oilseed, showing a beneficial effect of AMF in these plants. The achievement of this effective beneficial association among mycorrhizas and rapeseed plants opens new opportunities in agribiotechnology for the use of AMF as biofertilizers in Brassicaceae crops with potential application in medical, animal and industrial biotechnology.

Role of rhizospheric microbial enzymes in plant growth promotion, antagonism, and sustainable agriculture: A review

Plant growth-promoting rhizobacteria (PGPR) offer eco-friendly alternatives to chemical fertilizers, promoting sustainable agriculture by enhancing soil fertility, reducing pathogens, and aiding in stress resistance. In agriculture, they play a crucial role in plant growth promotion through the production of agroactive compounds and extracellular enzymes to promote plant health and protection against phytopathogens. In the rhizosphere, diverse microbial interactions, including those with bacteria and fungi, influence plant health by production of antimicrobial compounds. The antagonism displayed by rhizobacteria plays a crucial role in shaping microbial communities and has potential applications in developing a natural and environmentally friendly approach to pest control. The rhizospheric microbes showcase their ecological importance and potential for biotechnological applications in the context of plant-microbe interactions. The extracellular enzymes produced by rhizospheric microbes like amylases, chitinases, glucanases, cellulases, proteases, and ACC deaminase contribute to plant processes and stress response emphasizing their importance in sustainable agriculture. Moreover, this review highlights the new paradigm including artificial intelligence (AI) in sustainable horticulture and agriculture as a harmonious interaction between ecological networks for promoting soil health and microbial diversity that leads to a more robust and self-regulating agricultural system for protecting the environment in the future. Overall, this review emphasizes microbial interactions and the role of rhizospheric microbial extracellular enzymes which is crucial for developing eco-friendly approaches to enhance crop production and soil health.

Bacillus Strains as Effective Biocontrol Agents Against Phytopathogenic Bacteria and Promoters of Plant Growth.

Modern crop production relies on the application of chemical pesticides and fertilizers causing environmental and economic challenges. In response, less environmentally impactful alternatives have emerged such as the use of beneficial microorganisms. These microorganisms, particularly plant growth-promoting bacteria (PGPB), have demonstrated their ability to enhance plant growth, protect against various stresses, and reduce the need for chemical inputs. Among the PGPB, Bacillus species have garnered attention due to their adaptability and commercial potential. Recent reports have highlighted Bacillus strains as biocontrol agents against phytopathogenic bacteria while concurrently promoting plant growth. We also examined Bacillus plant growth-promoting abilities in Arabidopsis thaliana seedlings. In this study, we assessed the potential of various Bacillus strains to control diverse phytopathogenic bacteria and inhibit quorum sensing using Chromobacterium violaceum as a model system. In conclusion, our results suggest that bacteria of the genus Bacillus hold significant potential for biotechnological applications. This includes developments aimed at reducing agrochemical use, promoting sustainable agriculture, and enhancing crop yield and protection.

Sulfonated cottonseed hydrolysates with adjustable amphiphilicity as environmental -Stress stable emulsifiers

Cottonseed protein isolate (CPI) is a valuable agro-industrial waste with potential biotechnological applications. However, inadequate stability in water due to its characteristic hinders its widespread use. Therefore, a new sulfonation modification approach was developed to improve the amphiphilicity and structural flexibility of CPI. Structural characterizations confirmed the successful incorporation of sulfonate groups with structural and conformational changes. This significantly unfolded molecular-chain, and improved amphiphilicity, flexibility, and surface-hydrophobicity while reducing pI (5.1–1.7), and molecular-weight (5745–2089 g/mol). The modified samples exhibited improved emulsification with higher amounts of absorbed proteins on the droplet interface, smaller droplet size, and a higher zeta-potential. Additionally, they possessed good emulsification ability under acidic conditions. The nano-emulsions exhibited long-term stability (≥70 days) under different environmental conditions, with excellent fluidity. This study contributes to understanding sulfonation as a viable approach for improving protein properties, thus, opening up new possibilities for their application and maximizing their economic benefits.

Characterization of melanin from Exophiala mesophila with the prospect of potential biotechnological applications.

Fungal melanin is an underexplored natural biomaterial of great biotechnological interest in different areas. This study investigated the physical, chemical, electrochemical, and metal-binding properties of melanin extracted from the metallotolerant black fungus Exophiala mesophila strain IRTA-M2-F10. Specific inhibitory studies with tricyclazole and biochemical profiling of whole cells by synchrotron radiation-based Fourier-transform infrared spectral microscopy (SR-FTIRM) were performed. An optimized extraction protocol was implemented, and purified fungal melanin was characterized using an array of spectrophotometric techniques (UV-Vis, FTIR, and EPR) and by cyclic voltammetry (CV) experiments. The metal-binding capacity of melanin extracts was also assessed by using Cr(VI) as a model heavy metal. Inhibitory studies indicated that 1,8-dihydroxynaphthalene may be the main precursor molecule of E. mesophila melanin (DHN-melanin). The biochemical characterization of fungal melanin extracts were benchmarked against those from two melanins comprising the precursor molecule L-3,4-dihydroxiphenylalanine (DOPA-melanin): extracts from the ink of the cephalopod Sepia officinalis and DOPA-melanin synthesized in the laboratory. The CV results of melanin extracts incubated with and without cell suspensions of the electroconductive bacterium Geobacter sulfurreducens were indicative of novel semiquinone/hydroquinone redox transformations specific for each melanin type. These interactions may play an important role in cation exchange for the adsorption of metals and in microbial interspecies electron transfer processes. The obtained results provided further evidence for the DHN-nature of E. mesophila melanin. The FTIR profiling of melanin extracts exposed to Cr(VI), compared to unexposed melanin, resulted in useful information on the distinct surface-binding properties of fungal melanin. The parameters of the Langmuir and Freundlicht isotherms for the adsorption of Cr(VI) were determined and compared to bibliographic data. Altogether, the inherent properties of fungal melanin suggest its promising potential as a biomaterial for environmental applications.

Heterologous expression of nattokinase in E. coli: Biochemical characterization and functional analysis of fibrin binding residues

Heterologous expression of nattokinase, a potent fibrinolytic enzyme, has been successfully carried out in various microorganisms. However, the successful expression of this enzyme as a soluble protein was not achieved in E. coli. This study delves into the expression of nattokinase in E. coli as a soluble protein followed by its biochemical characterization and functional analysis for fibrinolytic activity. E. coli BL21C41 and pET32a vector host strain with pGro7 protein chaperone induced with IPTG at 16 °C 180 rpm for 16 h enabled the production of recombinant nattokinase in soluble fraction. Enzymatic assays demonstrated its protease activity, while characterization revealed optimal catalytic conditions at 37 °C and pH 8.0, with remarkable stability over a broad pH range (6.0–10.0) and up to 50 °C. The kinetic constants were determined as follows: Km = 25.83 ± 3.43 μM, Vmax = 62.91 ± 1.68 μM/s, kcat = 38.45 ± 1.06 s−1, and kcat/Km = 1.49 × 106 M−1 s−1. In addition, the fibrinolytic activity of NK, quantified by the fibrin plate hydrolysis assay was 1038 ± 156 U/ml, with a corresponding specific activity of 1730 ± 260 U/mg and the assessment of clot lysis time on an artificial clot (1 mg) was found to be 51.5 ± 2.5 min unveiling nattokinase's fibrinolytic potential. Through molecular docking, a substantial binding energy of −6.46 kcal/mol was observed between nattokinase and fibrin, indicative of a high binding affinity. Key fibrin binding residues, including Ser300, Leu302, and Asp303, were identified and confirmed. These mutants affected specifically the fibrin binding and not the proteolytic activity of NK. This comprehensive study provides crucial conditions for the expression of protein in soluble form in E. coli and biochemical properties paving the way for future research and potential applications in medicine and biotechnology.

Screening of Bacillus spp. bacterial endophytes for protease production, and application in feather degradation and bio-detergent additive

Research on proteases and secondary metabolites from endophytes is an area that requires attention from researchers. In this study, proteases from Bacillus sp. strain MHSD16 and Bacillus sp. strain MHSD17 endophytes were characterised, and their potential biotechnological applications were investigated. Optimum protease production was achieved when isolates were grown in media containing (g/L): glucose 10g, casein 5g, yeast extract 5g, KH2PO4 2g, Na2CO3 10g at pH 9. The crude protease extracts were active in alkaline environments, thus referred to as alkaline proteases with optimal pH of 10. Additionally, Bacillus sp. strain MHSD 16 and Bacillus sp. strain MHSD17 proteases were active at high temperatures, with optimum enzyme activity at 50 °C. Thermostability profiles of these proteases showed that the enzymes were highly stable between (40–60 °C), maintaining over 85 % stability after 120 min incubation at 60 °C. Furthermore, the enzymes were stable and compatible with various household and laundry detergents. In the presence of commercial laundry detergent, OMO® 68 % and 72 % activity was retained for Bacillus sp. strain MHSD16 and Bacillus sp. strain MHSD17, respectively, while 67 % and 68 % activity were retained in the presence of Sunlight®. The potential application for use in detergents was investigated through the removal of blood stains with the crude alkaline extracts displaying efficient stain removal abilities. Feather degradation was also investigated and Bacillus sp. MHSD17 exhibited feather keratin degrading properties more effectively than Bacillus sp. MHSD16.

The Role of Microbial Diversity in Lignocellulosic Biomass Degradation: A Biotechnological Perspective

AbstractLignocellulosic biomass, such as plant residues and agricultural waste, holds immense potential as a renewable resource for the production of biofuels, chemicals, and animal feed. However, the efficient degradation of lignocellulose into fermentable sugars remains a significant challenge. Recent research has highlighted the critical role of microbial diversity in lignocellulosic biomass degradation, offering new insights from a biotechnological perspective. The comprehension and utilization of microbial diversity are crucial for developing efficient biotechnological strategies for lignocellulosic biomass degradation. By uncovering the intricate relationships between microbial communities and their enzymatic machinery, researchers can optimize degradation processes, enhance biofuel production, and contribute to a more sustainable bio‐based economy. Microorganisms, including bacteria, fungi, and archaea, possess diverse enzymatic capabilities, allowing them to secrete a plethora of lignocellulolytic enzymes. Microbial organisms inhabiting extreme environments, such as the rumen, hot and cold springs, deep sea trenches, and acidic and alkaline pH environments, exhibit significant potential in generating enzymes, including hemicellulolytic and lignocellulolytic enzymes, which possess superior biochemical properties essential for industrial bioconversion applications. This review explores the ability of lignocellulosic enzymes from microbial sources to efficiently break down the lignocellulosic biomass and their potential applications in industrial biotechnology.

Direct Degradation of Fresh and Dried Macroalgae by Agarivorans albus B2Z047.

Marine macroalgae are increasingly recognized for their significant biological and economic potential. The key to unlocking this potential lies in the efficient degradation of all carbohydrates from the macroalgae biomass. However, a variety of polysaccharides (alginate, cellulose, fucoidan, and laminarin), are difficult to degrade simultaneously in a short time. In this study, the brown alga Saccharina japonica was found to be rapidly and thoroughly degraded by the marine bacterium Agarivorans albus B2Z047. This strain harbors a broad spectrum of carbohydrate-active enzymes capable of degrading various polysaccharides, making it uniquely equipped to efficiently break down both fresh and dried kelp, achieving a hydrolysis rate of up to 52%. A transcriptomic analysis elucidated the presence of pivotal enzyme genes implicated in the degradation pathways of alginate, cellulose, fucoidan, and laminarin. This discovery highlights the bacterium's capability for the efficient and comprehensive conversion of kelp biomass, indicating its significant potential in biotechnological applications for macroalgae resource utilization.

Insights into Early Phases of Phycocyanin Crystal Formation via SONICC Spectroscopy

This research delves into the early nucleation stages of phycocyanin, a protein pivotal for its fluorescent properties and crystalline stability and holding considerable potential for biotechnological applications. The paper contrasts traditional crystallization methods with the innovative Langmuir–Blodgett nanotemplate approach, aiming to enhance molecular assembly and nucleation processes. The study employs Langmuir–Blodgett nanotemplates alongside second-order nonlinear imaging of chiral crystal (SONICC) spectroscopy. This combination is designed to orderly organize phycocyanin molecules and provide a sensitive visualization of early-stage crystal formation, capturing the intricate dynamics of protein crystallization. The experiments were conducted under controlled conditions, where surface pressure was maintained at 26 mN/m and barrier speed at 70 cm/min to optimize the monolayer formation at the air–water interface. The Langmuir–Blodgett method, compared to traditional vapor diffusion techniques, shows improvements in the uniformity and efficiency of nucleation. The sensitivity of SONICC spectroscopy significantly enhances the visualization of the nucleation process, revealing a more structured and uniform crystalline assembly in the early stages of formation. This method demonstrates a substantial improvement in nucleation dynamics, leading to a more orderly growth process and potentially larger, well-ordered crystals. Integrating Langmuir–Blodgett nanotemplates with SONICC spectroscopy offers a significant step in understanding protein crystallization processes with insights into the nucleation and growth of protein crystals and broad implications for refining crystallography methodologies of protein-based biomaterials, contributing to the advancement of structural biology and materials science.

In silico biotechnological potential of Bacillus sp. strain MHSD_37 bacterial endophyte

BackgroundEndophytic bacteria possess a range of unique characteristics that enable them to successfully interact with their host and survive in adverse environments. This study employed in silico analysis to identify genes, from Bacillus sp. strain MHSD_37, with potential biotechnological applications.ResultsThe strain presented several endophytic lifestyle genes which encode for motility, quorum sensing, stress response, desiccation tolerance and root colonisation. The presence of plant growth promoting genes such as those involved in nitrogen fixation, nitrate assimilation, siderophores synthesis, seed germination and promotion of root nodule symbionts, was detected. Strain MHSD_37 also possessed genes involved in insect virulence and evasion of defence system. The genome analysis also identified the presence of genes involved in heavy metal tolerance, xenobiotic resistance, and the synthesis of siderophores involved in heavy metal tolerance. Furthermore, LC-MS analysis of the excretome identified secondary metabolites with biological activities such as anti-cancer, antimicrobial and applications as surfactants.ConclusionsStrain MHSD_37 thereby demonstrated potential biotechnological application in bioremediation, biofertilisation and biocontrol. Moreover, the strain presented genes encoding products with potential novel application in bio-nanotechnology and pharmaceuticals.

Electrochemical Impedance Spectroscopy for the Sensing of the Kinetic Parameters of Engineered Enzymes.

The study presents a promising approach to enzymatic kinetics using Electrochemical Impedance Spectroscopy (EIS) to assess fundamental parameters of modified enteropeptidases. Traditional methods for determining these parameters, while effective, often lack versatility and convenience, especially under varying environmental conditions. The use of EIS provides a novel approach that overcomes these limitations. The enteropeptidase underwent genetic modification through the introduction of single amino acid modifications to assess their effect on enzyme kinetics. However, according to the one-sample t-test results, the difference between the engineered enzymes and hEKL was not statistically significant by conventional criteria. The kinetic parameters were analyzed using fluorescence spectroscopy and EIS, which was found to be an effective tool for the real-time measurement of enzyme kinetics. The results obtained through EIS were not significantly different from those obtained through traditional fluorescence spectroscopy methods (p value >> 0.05). The study validates the use of EIS for measuring enzyme kinetics and provides insight into the effects of specific amino acid changes on enteropeptidase function. These findings have potential applications in biotechnology and biochemical research, suggesting a new method for rapidly assessing enzymatic activity.

Optogenetic Control of Bacterial Cell-Cell Adhesion Dynamics: Unraveling the Influence on Biofilm Architecture and Functionality.

The transition of bacteria from an individualistic to a biofilm lifestyle profoundly alters their biology. During biofilm development, the bacterial cell-cell adhesions are a major determinant of initial microcolonies, which serve as kernels for the subsequent microscopic and mesoscopic structure of the biofilm, and determine the resulting functionality. In this study, the significance of bacterial cell-cell adhesion dynamics on bacterial aggregation and biofilm maturation is elucidated. Using photoswitchable adhesins between bacteria, modifying the dynamics of bacterial cell-cell adhesions with periodic dark-light cycles is systematic. Dynamic cell-cell adhesions with liquid-like behavior improve bacterial aggregation and produce more compact microcolonies than static adhesions with solid-like behavior in both experiments and individual-based simulations. Consequently, dynamic cell-cell adhesions give rise to earlier quorum sensing activation, better intermixing of different bacterial populations, improved biofilm maturation, changes in the growth of cocultures, and higher yields in fermentation. The here presented approach of tuning bacterial cell-cell adhesion dynamics opens the door for regulating the structure and function of biofilms and cocultures with potential biotechnological applications.