A procedure resulting in a highly purified preparation of bacterial luciferase with a high specific activity towards FMNH2 and NADH was developed. Using SDS-electrophoresis in polyacrylamide gel, it was shown that the enzyme has a subunit composition and that its monomers do not contain the luciferase activity. The use of the obtained preparation for determining the content of NADH and FMN by the bioluminescent method allowed to increase its sensitivity up to 10(-18) and 10(-17) moles, respectively.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Thymol as a Novel Plant-Derived Antibacterial Agent for Suppressing Xanthomonas citri pv. malvacearum in Cotton.
Curr Microbiol
January 2025
Department of Plant Pathology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan.
Xanthomonas citri pv. malvacearum (Xcm) associated with bacterial blight disease is a significant and widespread pathogen affecting cotton worldwide. The excessive use of harmful chemicals to control plant pathogens has exerted a negative impact on environmental safety.
View Article and Find Full Text PDFDesign and Characterization of a Gene-Encoding DNA Nanoparticle in a Cell-Free Transcription-Translation System.
ACS Appl Nano Mater
June 2024
Department of Chemistry, College of Arts and Sciences, Case Western Reserve University, Cleveland, Ohio 44106, United States.
DNA nanotechnology has made initial progress toward developing gene-encoded DNA origami nanoparticles (NPs) that display potential utility for future gene therapy applications. However, due to the challenges involved with gene delivery into cells including transport through the membrane, intracellular targeting, and inherent expression of nucleases along with interference from other active proteins, it can be difficult to more directly study the effect of DNA NP design on subsequent gene expression. In this work, we demonstrate an approach for studying the expression of gene-encoding DNA origami NPs without the use of cells.
View Article and Find Full Text PDFMechanisms and applications of bacterial luciferase and its auxiliary enzymes.
Arch Biochem Biophys
January 2025
Department of Biochemistry and Center of Excellent in Protein Structure & Function, Faculty of Science, Mahidol University, Bangkok, 14000, Thailand. Electronic address:
Bacterial luciferase (LuxAB) catalyzes the conversion of reduced flavin mononucleotide (FMNH⁻), oxygen, and a long-chain aldehyde to oxidized FMN, the corresponding acid and water with concomitant light emission. This bioluminescence reaction requires the reaction of a flavin reductase such as LuxG (in vivo partner of LuxAB) to supply FMNH⁻ for the LuxAB reaction. LuxAB is a well-known self-sufficient luciferase system because both aldehyde and FMNH⁻ substrates can be produced by the associated enzymes encoded by the genes in the lux operon, allowing the system to be auto-luminous.
View Article and Find Full Text PDFScreening to isolate Bacillus subtilis mutants with enhanced NADPH levels.
J Gen Appl Microbiol
January 2025
Department of Science, Technology and Innovation, Kobe University.
As the first step toward understanding how NADPH levels are regulated in Bacillus subtilis, we sought to obtain mutant strains with enhanced NADPH levels. Our previous study demonstrated that in a strain of B. subtilis expressing bacterial luciferase derived from Photorhabdus luminescens, artificially enhancing NADPH levels enhanced luciferase luminescence in the colonies.
View Article and Find Full Text PDFEngineering a thermophilic luciferase variant from Photuris pennsylvanica into a mesophilic-like enzyme for expanded applications potential.
Int J Biol Macromol
January 2025
Anhui Academy of Medical Sciences, Anhui Medical College, Hefei, China. Electronic address:
Luciferase, known for its exceptional catalytic bioluminescent properties, has been widely utilized in diverse applications within biotechnology and medical research. Currently, enhancing thermostability and catalytic activity is a primary focus for optimizing luciferase modifications to further expand its detection range and accuracy. This study revealed a highly thermostable luciferase variant from Photuris pennsylvanica, Ppe146-1H2, which inherently exhibits thermophilic enzyme characteristics that are not conducive for optimal catalytic performance in practical applications.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!