Alcohol oxidase (EC 1.1.3.13; AOX) is a flavoprotein that catalyzes the oxidation of primary short-chain alcohols to corresponding carbonyl compounds with a concomitant release of hydrogen peroxide. It is a key enzyme of methanol metabolism in methylotrophic yeasts, catalyzing the first step of methanol oxidation to formaldehyde.Here we describe the isolation and purification of AOX from the thermotolerant methylotrophic yeast Ogataea (Hansenula) polymorpha, and using this enzyme in enzymatic assay of ethanol, simultaneous analysis of methanol and formaldehyde, and in construction of amperometric biosensors selective to primary alcohols and formaldehyde.
We propose a hybrid electrocatalytic-bioelectrocatalytic reaction cascade integrated on a gas diffusion electrode for CO2 reduction under selective formation of methanol. Ag-Bi2O3 selectively reduces gaseous CO2 to formate at neutral pH conditions. A subsequent enzymatic cascade comprising formaldehyde dehydro-genase and alcohol dehydrogenase, which are both nicotinamide adenine dinucleotide (NAD)-dependent, further reduce formate sequentially to formaldehyde and methanol.
The vanillyl alcohol oxidase/p-cresol methylhydroxylase (VAO/PCMH) flavoprotein family comprises a broad spectrum of enzymes capable of catalyzing the oxidative bioconversions of various substrates. Among them, pinoresinol hydroxylase (PinH) from the 4-alkylphenol oxidizing subgroup initiates the oxidative degradation of (+)-pinoresinol, a lignan important for both lignin structure and plant defense. In this study, we present a detailed biochemical and structural characterization of PinH from Pseudomonas sp.
Department of Biochemistry and Biotechnology, Institute of Biological Sciences, Maria Curie-Skłodowska University, Akademicka 19 St., 20-033 Lublin, Poland.
Intracellular alcohol oxidase (AOX) was isolated from the basidiomycetous white rot fungus FCL139. The enzyme was semi-purified (13-fold) using two-step chromatography with 30% activity recovery. The identity of the protein was confirmed by LC-MS/MS analysis, and its MW (72 kDa) and pI (6.
Methylotrophic yeast is a useful host for producing heterologous proteins using the unique and strong alcohol oxidase 1 (AOX1) promoter, which is induced by methanol and repressed by various carbon sources. However, methanol is preferably avoided in industrial-scale fermentation given its toxicity, flammability, and explosiveness. To develop a protein production system under reduced methanol supply conditions, we attempted to characterize the AOX1 promoter induction activity by comparing derepression conditions with methanol induction conditions.
Alcohol oxidase (AOx) enzymes are important for industrial applications due to their ability to oxidize various alcohols without needing extra co-factors, and this study focuses on enhancing the understanding of recombinant AOx from MTCC6324 using advanced computational methods.
The research utilizes technologies like artificial intelligence (AlphaFold), molecular docking, and molecular dynamics simulations to analyze the enzyme's structure and functions, revealing significant stability and binding interactions with substrates like cinnamyl alcohol.
The findings highlight a unique substrate sequestration mechanism where cinnamyl alcohol temporarily binds to the enzyme before detaching, providing valuable insights for the future design of biocatalytic processes.
