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Journal of Biotechnology Feb 2024l-Lactate oxidase has important applications in biosensing and finds increased use in biocatalysis. The enzyme has been characterized well, yet its immobilization has...
l-Lactate oxidase has important applications in biosensing and finds increased use in biocatalysis. The enzyme has been characterized well, yet its immobilization has not been explored in depth. Here, we studied immobilization of Aerococcus viridansl-lactate oxidase on porous carriers of variable matrix material (polymethacrylate, polyurethane, agarose) and surface functional group (amine, Ni-loaded nitrilotriacetic acid (NiNTA), epoxide). Carrier activity (A) and immobilized enzyme effectiveness (ɳ) were evaluated in dependence of protein loading. Results show that efficient immobilization (A: up to 1450 U/g carrier; ɳ: up to 65%) requires a hydrophilic carrier (agarose) equipped with amine groups. The value of ɳ declines sharply as A increases, probably due to transition into diffusional regime. Untagged l-lactate oxidase binds to NiNTA carrier similarly as N-terminally His-tagged enzyme. Lixiviation studies reveal quasi-irreversible enzyme adsorption on NiNTA carrier while partial release of activity (≤ 25%) is shown from amine carrier. The desorbed enzyme exhibits the same specific activity as the original l-lactate oxidase. Collectively, our study identifies basic requirements of l-lactate oxidase immobilization on solid carrier and highlights the role of ionic interactions in enzyme-surface adsorption.
Topics: Aerococcus; Sepharose; Mixed Function Oxygenases; Enzymes, Immobilized; Amines
PubMed: 38280467
DOI: 10.1016/j.jbiotec.2024.01.011 -
Foods (Basel, Switzerland) Jul 2023In order to investigate and develop functional foods of marine origin with hypoglycemic activity, polysaccharide-Zn(II) (EZ) complex was first prepared by marine...
In order to investigate and develop functional foods of marine origin with hypoglycemic activity, polysaccharide-Zn(II) (EZ) complex was first prepared by marine resourced polysaccharide (EP) and ZnSO and their anti-diabetes activities against high-sugar and high-fat-induced diabetic mice were evaluated. The detailed structural characterization of EZ was elucidated by UV-Vis spectroscopy, infrared spectroscopy, and monosaccharide composition determination. The pharmacological research suggests that EZ has a potent hypoglycemic effect on high-sugar and high-fat-induced diabetic mice by inhibiting insulin resistance, improving dyslipidemia, decreasing inflammatory status, repairing pancreas damage, as well as activating the IRS/PI3K/AKT signaling pathway and regulating GLUT2 gene expression. At the same time, microbiota analysis indicates that a high dose of EZ could enhance the abundance of dominant species, such as , , , , and , in intestinal microbiota distribution. Thus, EZ could be considered as a potential candidate for developing an ingredient of functional foods for Zn(II) supplements with hypoglycemic activity.
PubMed: 37569125
DOI: 10.3390/foods12152854 -
International Journal of Molecular... Nov 2023() is a swine pathogen that can cause sepsis, meningitis, endocarditis, and other infectious diseases; it is also a zoonotic pathogen that has caused a global surge in...
() is a swine pathogen that can cause sepsis, meningitis, endocarditis, and other infectious diseases; it is also a zoonotic pathogen that has caused a global surge in fatal human infections. The widespread prevalence of multidrug-resistant strains and the decline in novel antibiotic candidates have necessitated the development of alternative antimicrobial agents. In this study, AVPL, the () phage lysin, was found to exhibit efficient bactericidal activity and broad lytic activity against multiple serotypes of . A final concentration of 300 μg/mL AVPL reduced counts by 4-4.5 log10 within 1 h in vitro. Importantly, AVPL effectively inhibited 48 h biofilm formation and disrupted preformed biofilms. In a mouse model, 300 μg/mouse AVPL protected 100% of mice from infection following the administration of lethal doses of multidrug-resistant type 2 (SS2) strain SC19, reduced the bacterial load in different organs, and effectively alleviated inflammation and histopathological damage in infected mice. These data suggest that AVPL is a valuable candidate antimicrobial agent for treating infections.
Topics: Animals; Swine; Humans; Mice; Bacteriophages; Aerococcus; Streptococcus suis; Streptococcal Infections; Bacteremia; Disease Models, Animal
PubMed: 38068990
DOI: 10.3390/ijms242316670 -
Emerging Infectious Diseases Aug 2023We report a novel Globicatella species causing extensive soft tissue infection in a man bitten by a stray domestic cat in the United Kingdom. We identified this...
We report a novel Globicatella species causing extensive soft tissue infection in a man bitten by a stray domestic cat in the United Kingdom. We identified this bacterium by 16S rRNA gene sequencing, whole-genome sequencing, and biochemical profiling and determined antimicrobial drug susceptibility.
Topics: Animals; Cats; Gram-Positive Bacterial Infections; RNA, Ribosomal, 16S; Soft Tissue Infections; Aerococcaceae; Bacteria
PubMed: 37486350
DOI: 10.3201/eid2908.221770 -
Biomedicine & Pharmacotherapy =... Jun 2024There is a growing evidence suggesting the association of vitamin D deficiency (VDD) and cognitive impairment. In this study we evaluated the possible involvement of gut...
There is a growing evidence suggesting the association of vitamin D deficiency (VDD) and cognitive impairment. In this study we evaluated the possible involvement of gut microbiota in the cognitive impairments mediated by VDD and investigated the effects of pharmacological treatment with the oxazoline derivative of the aliamide palmitoylethanolamide, 2-Pentadecyl-2-oxazoline (PEA-OXA). Mice were submitted to behavioural, biochemical and electrophysiological analysis to assess whether their vitamin D status affected cognitive performance together with gut microbiota composition. In VDD mice we found cognitive malfunctioning associated with reduced neuroplasticity, indicated by impaired long term potentiation, and neuroinflammation at the hippocampal level. Importantly, PEA-OXA counteracted the cognitive impairments and modified the biochemical and functional changes induced by VDD. Additionally, PEA-OXA treatment enhanced gut microbiota diversity, which tended to be decreased by VDD only in female mice, elevated the relative abundance of lactic and butyric acid-producing families, i.e. Aerococcaceae and Butyricicoccaceae, and reversed the VDD-induced decrease of butyrate-producing beneficial genera, such as Blautia in female mice, and Roseburia in male mice. These data provide novel insights for a better understanding of the cognitive decline induced by VDD and related gut dysbiosis and its potential therapeutic treatment.
Topics: Animals; Gastrointestinal Microbiome; Cognitive Dysfunction; Male; Female; Mice; Vitamin D Deficiency; Hippocampus; Mice, Inbred C57BL; Ethanolamines; Dysbiosis; Amides; Cognition; Disease Models, Animal
PubMed: 38670046
DOI: 10.1016/j.biopha.2024.116600 -
Clinical Infectious Diseases : An... Oct 2023
Topics: Humans; Aerococcus; Endocarditis, Bacterial; Endocarditis; Heart; Communicable Diseases
PubMed: 37382039
DOI: 10.1093/cid/ciad393 -
Microbiology Spectrum Jun 2024Bacterial isolates from the human urinary microbiome have been extensively studied for their antibiotic resistance; however, little work has been done on those isolates...
UNLABELLED
Bacterial isolates from the human urinary microbiome have been extensively studied for their antibiotic resistance; however, little work has been done on those isolates that are difficult to grow . This study was designed to qualify a serum-based medium, New York City Broth III (NYCIII), and a broth microdilution method to determine the antibiotic susceptibility of previously underreported or undescribed microbes that have a difficult time growing in standard Mueller-Hinton broth. Here, we demonstrate that NYCIII microbroth dilution can be an effective method for the determination of antibiotic susceptibility of species found in the human urinary microbiome. We show that this method serves well to characterize fastidious and anaerobic urinary microbes that have no Clinical and Laboratory Standards Institute (CLSI) guidelines, including several in the families , , or . Previous studies using expanded quantitative urine culture reveal that urine samples from clinical patients are commonly polymicrobial in composition. Thus, we test whether NYCIII can serve as a viable harmonized medium, capable of supporting antibiotic susceptibility testing in a range of fastidious, non-fastidious, and anaerobic urinary microbes. We propose this methodology to be standardized comparable to CLSI standards to allow for resistance testing in uncharacterized urinary bacteria.
IMPORTANCE
Antibiotic susceptibilities of fastidious and anaerobic bacteria of the human urinary microbiome are largely underreported due to difficulty in growing them in the lab environment. The current standard medium, Muller-Hinton broth, has difficulty supporting the growth of many of these species, leaving microbiologists without a standardized method. To address this need, this study offers a methodology to survey susceptibilities in a high-throughput manner of these understudied microbes with a proposed harmonized medium, NYCIII, which is capable of supporting the growth of both fastidious and non-fastidious urinary microbes. Broader standardization of this method can allow for the development of antibiotic-resistant breakpoints of the many uncharacterized urinary microbes.
Topics: Humans; Microbial Sensitivity Tests; Anti-Bacterial Agents; Microbiota; Bacteria, Anaerobic; Urine; Urinary Tract Infections; Bacteria; Culture Media
PubMed: 38709058
DOI: 10.1128/spectrum.00314-24