Flavin mononucleotide cofactor

Chang FC, Swenson RP(1999)The midpoint potentials for the oxidized-semiquinone couple for Gly57 mutants of the Clostridium beijerinckii flavodoxin correlate with changes in the hydrogen-bonding interaction with the proton on N(5) of the reduced flavin mononucleotide cofactor as measured by NMR chemical shift temperature dependencies. Biochemistry 38 7168 7176... 

There are two interesting features to the reactiori. First, HLADH accomplishes two oxidations, to the hemiacetal and thence to the lactone. Second, as in the natural system (a horse), the four hydrogens from the substrate end up on the flavin mononucleotide cofactor (FMN). The NAD is recycled, and is therefore only needed in catalytic amount. 

Figure 8 Illustration of the ensemble of 10 E. coli Flavodoxin structures obtained from homology modeling using distance geometry, superimposed on the crystal structure (heavy line) so as to minimize the coordinate differences to the alpha carbons in residues 4-170. Only the heavy backbone and aromatic sidechain atoms are shown, together with those of the flavin mononucleotide cofactor (lower left)...

Flavin Mononucleotide (FMN) (C17H21N409P) is a phosphoric ester of riboflavin that constitutes the cofactor of various flavoproteins. 

All NOS isoforms utilize L-arginine as the substrate, and molecular oxygen and reduced nicotinamide adenine dinucleotide phosphate (NADPH) as cosubstrates. Flavin adenine dinucleotide (FMN), flavin mononucleotide (FAD), and (6R)-5,6,7,8-tetrahydro-L-biopterin (BH4) are cofactors of the enzyme. All NOS isoforms contain heme and bind calmodulin. In nNOS and eNOS,... 

Flavoprotein enzymes contain flavin mononucleotide (FMN) or flavin adenine dinucleotide (FAD) as prosthetic groups. FMN and FAD are formed in the body from the vitamin riboflavin (Chapter 45). FMN and FAD are usually tighdy—but not covalendy—bound to their respecdve apoenzyme proteins. Metalloflavopro-teins contain one or more metals as essential cofactors. 

Since long retention times are often applied in the anaerobic phase of the SBR, it can be concluded that reduction of many azo dyes is a relatively a slow process. Reactor studies indicate that, however, by using redox mediators, which are compounds that accelerate electron transfer from a primary electron donor (co-substrate) to a terminal electron acceptor (azo dye), azo dye reduction can be increased [39,40]. By this way, higher decolorization rates can be achieved in SBRs operated with a low hydraulic retention time [41,42]. Flavin enzyme cofactors, such as flavin adenide dinucleotide, flavin adenide mononucleotide, and riboflavin, as well as several quinone compounds, such as anthraquinone-2,6-disulfonate, anthraquinone-2,6-disulfonate, and lawsone, have been found as redox mediators [43—46]. 

Flavin mononucleotide (FMN), 25 796 cofactor regeneration using, 3 673 Flaviviruses, 3 137-138 Flavones, microwave-assisted synthesis of, 16 575... 

All bacteria where nitrate ester degradation has been characterized have very similar enzymes. The enzymes eatalyze the nicotinamide cofactor-dependent reductive eleavage of nitrate esters that produces alcohol and nitrite. Purification of the PETN reduetase from Enterobacter cloacae yielded a monomerie protein of around 40 kilo Daltons, which required NADPH as a co-faetor for aetivity. Similar enzymes were responsible for the nitrate ester-degrading activity in Agrobacterium radiobacter (Snape et al. 1997) - nitrate ester reductase - and in the strains of Pseudomonas fluorescens and Pseudomonas putida (Blehert et al. 1999) - xenobiotic reduetases . All utilize a non-covalently bound flavine mononucleotide as a redox eofactor. 

The oxidation state of thiazolines and oxazolines can be adjusted by additional tailoring enzymes. For instance, oxidation domains (Ox) composed of approximately 250 amino acids utilize the cofactor FMN (flavin mononucleotide) to form aromatic oxazoles and thiazoles from oxazolines and thiazolines, respectively. Such domains are likely utilized in the biosynthesis of the disorazoles, " diazonimides, bleomycin, and epothiolone. The typical domain organization for a synthetase containing an oxidation domain is Cy-A-PCP-Ox however, in myxothiazol biosynthesis one oxidation domain is incorporated into an A domain. Alternatively, NRPSs can utilize NAD(P)H reductase domains to convert thiazolines and oxazolines into thiazolidines and oxazolidines, respectively. For instance, PchC is a reductase domain from the pyochelin biosynthetic pathway that acts in trans to reduce a thiazolyinyl-Y-PCP-bound intermediate to the corresponding thiazolidynyl-Y-PCP. ... 

Riboflavin (from the Latin flavus, yellow) serves in the metabolism as a component of the redox coenzymes flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD see p. 104). As prosthetic groups, FMN and FAD are cofactors for various oxidoreductases (see p. 32). No specific disease due to a deficiency of this vitamin is known. 

The C-terminal portion of the NOS protein closely resembles to cytochrome P-450 reductase, possesses many of the same cofactor binding sites, and basically performs the same functions. Consequently, this portion is often referred to as the reductase domain. At the extreme C-terminus is an NADPH binding region, which is conserved in all NOS and aligns perfectly with that of cytochrome P-450 reductase. The NADPH binding site is followed, in turn, by flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) consensus sequences. 

Ochoa reported that malic enzyme from L. plantarum was NAD and not NADP specific. The malic enzyme of cauliflower bud mitochondria (31) is NAD and NADP specific, with NAD being the preferred cofactor. Both the malo-lactic activity and NADH producing activity of the Leuconostoc oenos system (6,7, 8) was strictly NAD specific. Nicotinamide-adenine dinucleotide phosphate, flavin adenine dinucleotide, and flavin mononucleotide could not substitute in either of these activities. 

In 1989, BH4 was found to be a cofactor for nitric oxide synthase (NOS) [ 126, 127]. BH4 is also involved in dimerization of NOS, as NOS is catalytically active in a homodimer structure. Three isoforms of NOS exist neuronal NOS (NOS 1), inducible NOS (NOS 2) and endothelial NOS (NOS 3). BH4 is essential for all NOS isoforms. The NOS isoforms share approximately 50-60% sequence homology. Each NOS polypeptide is comprised of oxygenase and reductase domains. An N-terminal oxygenase domain contains iron protoporphyrin IX (heme), BH4 and an arginine binding site, and a C-terminal reductase domain contains flavin mononucleotide (FMN), and a reduced nicotin-amide adenine dinucleotide phosphate (NADPH) binding site. 

A model of a flavin-based redox enzyme was prepared.[15] Redox enzymes are often flavoproteins containing flavin cofactors flavin adenine dinucleotide (FAD) or flavin mononucleotide (FMN). They mediate one- or two-electron redox processes at potentials which vary in a range of more than 500 mV. The redox properties of the flavin part must be therefore tuned by the apoenzyme to ensure the specific function of the enzyme. Influence by hydrogen bonding, aromatic stacking, dipole interactions and steric effects have been so far observed in biological systems, but coordination to metal site has never been found before. Nevertheless, the importance of such interactions for functions and structure of other biological molecules make this a conceivable scenario. 

Flavoenzymes are widespread in nature and are involved in many different chemical reactions. Flavoenzymes contain a flavin mononucleotide (FMN) or more often a flavin adenine dinucleotide (FAD) as redox-active prosthetic group. Both cofactors are synthesized from riboflavin (vitamin B2) by microorganisms and plants. Most flavoenzymes bind the flavin cofactor in a noncovalent mode (1). In about 10% of aU flavoenzymes, the isoalloxazine ring of the flavin is covalently linked to the polypeptide chain (2, 3). Covalent binding increases the redox potential of the flavin and its oxidation power, but it may also be beneficial for protein stability, especially in flavin-deficient environments. 

Enzymatic cofactors, such as nicotinamide adenine dinucleotide (NADH), nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (EAD), flavin mononucleotide (EMN), and pyridoxal phosphate, are fluorescent and commonly found associated with various proteins where they are responsible for electron transport (see Fig. lb and Table 1). NADH and NADPH in the oxidized form are nonfluorescent, whereas conversely the flavins, FAD and EMN, are fluorescent only in the oxidized form. Both NADH and FAD fluorescence is quenched by the adenine found within their cofactor structures, whereas NADH-based cofactors generally remain fluorescent when interacting with protein structures. The fluorescence of these cofactors is often used to study the cofactors interaction with proteins as well as with related enzymatic kinetics (1, 9-12). However, their complex fluorescent characteristics have not led to widespread applications beyond their own intrinsic function. 

Some proteins contain other native fluorophores in addition to fluorescent amino acids. These include cofactors such as nicotinamide adenine dinucleotide (fluorescent in its reduced, NADH state) and flavin adenine dinucleotide (FAD). NADH is weakly fluorescent in water, but its fluorescence yield increases markedly on binding to a protein-binding site with an emission peak around 470 nm (3). FAD and flavin mononucleotide (FMN) are also fluorescent with an emission maximum around 520 nm, but fluorescence is quenched on binding to many flavoproteins (4). 

Whereas redox reactions on metal centres usually only involve electron transfers, many oxidation/reduction reactions in intermediary metabolism, as in the case above, involve not only electron transfer, but hydrogen transfer as well — hence the frequently used denomination dehydrogenase . Note that most of these dehydrogenase reactions are reversible. Redox reactions in biosynthetic pathways usually use NADPH as their source of electrons. In addition to NAD and NADP+, which intervene in redox reactions involving oxygen functions, other cofactors like riboflavin (in the form of flavin mononucleotide, FMN, and flavin adenine dinucleotide, FAD) (Figure 5.3) participate in the conversion of [—CH2—CH2— to —CH=CH—], as well as in electron transfer chains. In addition, a number of other redox factors are found, e.g., lipoate in a-ketoacid dehydrogenases, and ubiquinone and its derivatives, in electron transfer chains. 

Other Coenzymes and Cofactors.— The chemical synthesis of riboflavin phosphates and their acetyl derivatives has been reinvestigated. Riboflavin 4 -monophosphate (10) is an important contaminant of commercial flavin mononucleotide (FMN), and... 

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