Aldehydes bacterial bioluminescence

Fig. 2.1 Mechanism of the bacterial bioluminescence reaction. The molecule of FMNH2 is deprotonated at N1 when bound to a luciferase molecule, which is then readily peroxidized at C4a to form Intermediate A. Intermediate A reacts with a fatty aldehyde (such as dodecanal and tetradecanal) to form Intermediate B. Intermediate B decomposes and yields the excited state of 4a-hydroxyflavin (Intermediate C) and a fatty acid. Light (Amax 490 nm) is emitted when the excited state of C falls to the ground state. The ground state C decomposes into FMN plus H2O. All the intermediates (A, B, and C) are luciferase-bound forms. The FMN formed can be reduced to FMNH2 in the presence of FMN reductase and NADH.

The reported quantum yields of the long-chain aldehydes in the luminescence reaction catalyzed by P. fischeri luciferase are 0.1 for dodecanal with the standard I (Lee, 1972) 0.13 for decanal with the standard I (McCapra and Hysert, 1973) and 0.15-0.16 for decanal, dodecanal and tetradecanal with the standard III (Shimomura et al., 1972). Thus, the quantum yield of long-chain aldehydes in the bacterial bioluminescence reaction appears to be in the range of 0.10-0.16. 

Quantula (Dyakia), 180, 334 Quantum yield, xvi, 361, 362 aequorin, 104, 106, 110 aldehydes in bacterial bioluminescence, 36, 41 Chaetopterus photoprotein, 224 coelenterazine, 85, 143, 149 Cypridina luciferin, 69-71 definition, xvi, 361 Diplocardia bioluminescence, 242 firefly luciferin, 12 fluorescent compound F, 73 Latia luciferin, 190 pholasin, 197 PMs, 286... 

FORMATION OF H2O2 IN BACTERIAL BIOLUMINESCENCE REACTION WITH FLAVINMONONUCLEOTIDE ACTIVATED WITH N-METHYLIMIDAZOLE ON THE PHOSPHATE GROUP WITHOUT ADDITION OF THE EXOGENOUS ALDEHYDE... 

The bacterial bioluminescent reaction requires the contribution of the flavine mononucleotide (FMN) reduced form (FMNH), a long-chain aldehyde (tet-radecanal), and oxygen. All these molecules produce an oxidative reaction in the presence of bacterial luciferase (EC 1.14.14.3), giving tetradecanoic acid, FMN, wateq and hght emission under these conditions. 

The bacterial bioluminescence of Phofobacfer/ t/m phosphoreum results from the interaction of the enzyme luciferase, a flavin mononucleotide in its reduced form, and a long chain (eight carbons or more) aliphatic aldehyde. The presence of oxygen is vital to this interaction. 

An essential feature of bacterial bioluminescence is the requirement for long chain aldehyde (p. 156). Some bacteria, or certain mutants thereof, can use long-chain fatty acids, especially myristic acid [107] as precursors of the aldehydes. Thus a dark mutant of B. harveyi emits light when myristic acid is added. The light yield is proportional to the quantity of the acid added, down to 10 picomoles per ml. 

The possibility of isolating the components of the two above-reported coupled reactions offered a new analytical way to determine NADH, FMN, aldehydes, or oxygen. Methods based on NAD(P)H determination have been available for some time and NAD(H)-, NADP(H)-, NAD(P)-dependent enzymes and their substrates were measured by using bioluminescent assays. The high redox potential of the couple NAD+/NADH tended to limit the applications of dehydrogenases in coupled assay, as equilibrium does not favor NADH formation. Moreover, the various reagents are not all perfectly stable in all conditions. Examples of the enzymes and substrates determined by using the bacterial luciferase and the NAD(P)H FMN oxidoreductase, also coupled to other enzymes, are listed in Table 5. 

Production of light by certain marine bacteria. The general consensus is that light is produced when bacterial luciferase catalyzes the bioluminescent oxidation of FMNH2 and a long chain aldehyde by molecular oxygen. Volume 1(1,2). 

Most of bacterial biosensors are based on the operon luxCDABE that codes for the bacterial luciferase founded in the marine bacteria V. fischeri and V. harveyi, and for an essential aldehyde substrate that would otherwise have to be supplied exogenously. The cluster luxAB cassette codes for the luciferase whereas luxCDE encodes a fatty acid reductase complex. The latter enzymes are responsible for the synthesis of the long-chain aldehyde that is required as substrate in the bioluminescence reaction (Meighen and Dunlap, 1993 Hakkila et al., 2002). Luciferase catalyses the oxidation reaction of flavin mononucleotide (FMNH2). A long-chain (7 to 16 carbons) aldehyde is reduced in presence of oxygen by the aldehyde reductase. The outcome of the bioluminescent reaction can be expressed as follows ... 

Lin L, Szitmer R, Meighen E. Binding of flavin and aldehyde to the active site of bacterial luciferase. In Stanley P, Kricka L. eds. Bioluminescence Chemiluminescence Progress Current Applications. Singapore World Scientific, 2002 89-92. 

Bioluminescence in the sea is produced by bacterial luciferase, a two-component flavin hydroxylase system. Luciferase oxidizes long-chain aldehydes to the corresponding fatty acids using O2 and reduced FMN as substrates (Equation (19)). 

Bacterial L. has not yet been characterized. The corresponding luciferase produces luminescence in the presence of FMNH and straight chain aldehydes with more than 7 C-atoms. Structural elucidation of L. was delayed on account of their low natural concentration. 30,000 fireflies were required for the isolation of IS mg L., and 40,000 sea pansies yielded only O.S mg of the Renitla L. Many L. and synthetic analogs show a spontaneous luminescence in proton-free solvents, such as dimethyl sulfoxide, but the quantum yield is lower than in bioluminescence. See also Photoproteins. 

Holzman, T.F. and Baldwin, T.O., Reversible inhibition of the bacterial luciferase catalyzed bioluminescence reaction by aldehyde substrate kinetic mechanism and Ugand effects. Biochemistry, 22, 2838, 1983. 

Hastings, J.W., Tu, S.-C., Becvar, J.E., and Presswood, R.P., Bioluminesce from the reaction of FMN, hydrogen peroxide, and long chain aldehyde with bacterial luciferase, Photochem. Photobiol, 29, 383, 1979. 

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