Bacillus megaterium soluble

Hydroxylation of long-chain carboxylic acids, amides, and alcohols—but not the esters or the corresponding alkanes—at the w-l, (0-2, and (o-3 positions by a soluble enzyme system from Bacillus megaterium (Miura and Fulco 1975 Narhi and Fulco 1986,1987). 

Miura Y, AJ Fulco (1975) u-l, co-2, and co-3 hydroxylation of long-chain fatty acids, amides and alcohols by a soluble enzyme system from Bacillus megaterium. Biochim Biophys Acta 388 305-317. 

Buchanan, J. F., and Fulco, A. J. 1978. Formation of 9,10-Epoxypalmitate and 9,10-Dihydroxypalmitate from Palmitoleic Acid by a Soluble System from Bacillus megaterium. Biochem. Biophys. Res. Comm., 85,1254-1260. 

Miura, Y., and Fulco, A. J. 1975. Omega-1, Omega-2 and Omega-3 Hydroxylation of Long-Chain Fatty-Acids, Amides and Alcohols by a Soluble Enzyme-System from Bacillus megaterium. Biochim. Biophys. Acta, 388, 305-317. 

Ruettinger, R. T., and Fulco, A. J. 1981. Epoxidation of Unsaturated Fatty-Acids by a Soluble Cytochrome-P-450-Dependent System from Bacillus megaterium. J. Biol. Chem., 256, 5728-5734. 

These screens have been used to direct the evolution of cytochrome P450 BM-3, a soluble enzyme from Bacillus megaterium that contains its reductase and hydroxylase domains on a single polypeptide chain. P450 BM-3 primarily catalyzes the hydroxylation of fatty acids ( 12 to 18 carbons long) at the a>-1, a>-2, and m-3 positions, but also... 

The degradation of alkanoic acids by P-oxidation has been noted parenthetically above, but alternative pathways may occur. For example, the metabolism of hexanoic acid by strains of Pseudomonas sp. may take place by co-oxidation with subsequent formation of succinate and 2-tetrahydrofurany-lacetate as a terminal metabolite (Kunz and Weimer 1983). In a strain of Cory neb acterium sp., the specificities of the relevant catabolic enzymes are consistent with the production of dodecanedioic acid by co-oxidation of dode-cane but not of hexadecanedioic acid from hexadecane (Broadway et al. 1993). Hydroxylation at subterminal (co-1, co-2, and co-3) positions of carboxylic acids with chain lengths of 12 to 18—and less readily of the corresponding alcohols, but not the carboxylic acids or the alkanes—has been observed (Miura and Fulco 1975) for a soluble enzyme system from a strain of Bacillus megaterium. Whereas in this organism co-2 hydroxylation is carried out by a soluble cytochrome P-450 BM 3 (Narhi and Fulco 1987), co-hydroxylation in P. oleovorans that carries the OCT plasmid is mediated by a three-component hydroxylase that behaves like a cytoplasmic membrane protein (Ruettinger et al. 1974 Kok et al. 1989). 

Another Class II P-450 redox system that has been extensively studied is the cytosolic flavocytochrome P-450 BM3 from Bacillus megaterium. BM3 is the fusion of a soluble P-450 domain with CPR." " " The FAD of BM3 is reduced by NADPH the electrons are transferred to FMN and then finally to the substrate-bound P-450 domain." " BM3 is the fastest reported P-450 monooxygenase,with the rate of hydride transfer from NADPH to FAD and the rate of electron transfer from FMN to heme several-fold above the mammalian P-450 redox systems." The structure of the full-length protein has yet to be solved, but the structure of the FAD- and NADPH-binding domain has been determined." This domain closely resembles rat liver CPR and contains several conserved residues implicated in NADPH binding and flavin reduction. 

Narhi, L.O. and A.J. Fulco (1982). Phenobarbital induction of a soluble cytochrome P450 dependent fatty acid monooxygenase from Bacillus megaterium. J. Biol. Chem. 257, 2147-2150. 

Narhi LO, Fulco AJ (1982) Phenobarbital induction of a soluble cytochrome P-450-dependent fatty acid monooxygenase in Bacillus megaterium. J Biol Chem 257 2147-2150... 

Palmitoleic acid Soluble enzymes from Bacillus megaterium 93... 

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