Clostridium Transferase

In combination of this polymerase with purified propionyl-CoA transferase of Clostridium propionicum, a two-enzyme in vitro PHB biosynthesis system was established which allowed the PHB synthesis from (R)-hydroxybutyric acid as substrate [119]. In this way, the PHB synthesis was independent of the consumption of the expensive CoA, and hence PHA could be readily produced in a semipreparative-scale... 

In contrast to coenzyme Bi2, where the alkyl moiety serves purely in a catalytic role, the alkyl group of methyl cobamides (MeCba s) is utilized as a reagent by MeCba-dependent enzymes it is only the cobamide portion of the coenzyme that is catalytic. The cobamide-dependent methyl transferases have been reviewed [11,24-27,165], Three cobamide-dependent methyl transferases have been studied in some cases, more than one protein is required. The Bi2 proteins include methionine synthase (officially called 5-methyltetrahydrofolate-L-homocysteine-S-methyltransferase [HCM] EC 2.1.1.13) MeCba-dependent enzyme from Meth-anosarcina barkeri (MT 0 and the corrinoid/Fe-S protein from Clostridium ther-moaceticum. 

Moriishi et al., 1991 Moriishi et al., 1993). Whereas the gene for C3 from C. botulinum strain C468 encodes a protein of 211 amino acids (without signal peptide) with a molecular mass of 23546 Da (Popoff etal., 1990 Popoff etal., 1991), C3 from C. botulinum strain C 003-9 encodes a protein of 204 amino acids with a molecular mass of 23119 Da (Nemoto eta/., 1991) showing about 65% identity with the other C3 isoform. Moreover, C3-related transferases are produced by Clostridium limosum (C. limosum exoenzyme) (Just et al., 1992a),... 

Simpson LL, Stiles BG, Zepeda H etal. (1989) Production by Clostridium spiroforme of an iotalike toxin that possesses mono(ADP-ribosyl)transferase activity Identification of a novel class of ADP- ribosyltransferases. In Infect. Immun. 57 255-61 Stiles BG, Wilkens TD (1986) Purification and characterization of Clostridium perfringens iota toxin dependence on two nonlinked proteins for biological activity. In Infect. Immun. 54 683-8... 

Popoff MR, Rubin EJ, Gill DM, et al. (1988) Actin-specific ADP-rlbosyl-transferase produced by a Clostridium difficile strain. In Infect Immun. 56, 2299—2306. 

The best understood reactions are those catalyzed by ADP-ribosyl transferase components of certain bacterial toxins. These include, for example, toxins produced by Vibrio cholerae (the causative agent of cholera), pertussis toxin (from Bordetella pertussis, which causes whooping cough), diphtheria toxin, exotoxin A from Pseudomonas aeruginosa (an opportunistic pathogen), and toxins from Clostridium botulinum. 

Inactivation of Rho with Clostridium botulinum C3 ADP-ribosyl-transferase (C3 exoenzyme, which specifically ADP-ribosylates and inactivates Rho) inhibits neutrophil chemotaxis to N-formyl peptides [369, 443] without inhibiting the initiation of actin polymerization [94]. Microscopic examination of these cells reveals that they are unaffected in their abilities to polarize and extend pseudopodia, rather C3 exoenzyme inhibition of migration is associated with inhibition of rear release. 

Selmer, X, Willanzheimer, A., and Hetzel, M. (2002) Propionate CoA-transferase from Clostridium pro-pionicum cloning of the gene and identification of glutamate 324 at the active site. Eur. J. Biochem., 269, 372-380. 

Wiesenborn, D.P., Rudolph, F.B, and Papoutsakis, E.T. (1989) Coenzyme A transferase from Clostridium acetobutylicum ATCC 824 and its role in the uptake of acids. Appl. Environ. Microbiol, 55, 323-329. 

Pathway V uses succinic semialdehyde dehydrogenase (SucD), 4-hydroxybutyrate dehydrogenase (4hbD), and 4-hydroxybutyrate-CoA CoA transferase (OrfZ) to synthesize 4-hydroxybutyryl-CoA for forming 4-hydroxybutyrate-containing PHA. Pathway V was reported in Clostridium kluyveri (Valentin and Dennis 1997). 

FIGURE 9.2 Physiology of ABE fermentation metabolism of Clostridium acetobutylicum with the respective enzymes and products. CoA, coenzyme A Ldh, lactate dehydrogenase Pdc, pyruvate decarboxylase Pfor, pyruvate ferredoxin oxidoreductase Fdred, ferredoxin reduced Thl, thiolase Hbd, p-hydroxybutyryl-CoA dehydrogenase Crt, crotonase Bed, butyryl-CoA dehydrogenase Etf, electron transfer flavoprotein Pta, phosphotransacetylase Ack, acetate kinase Ptb, phosphotransbutyrylase Buk, butyrate kinase Ctf A/B, acetoacetyl-CoA acyl-CoA transferase Adc, acetoacetate decarboxylase AdhE, aldehyde/alcohol dehydrogenase Bdh, butanol dehydrogenase. 

Tummala SB, Junne SG, Papoutsakis ET. (2003a). Antisense RNA downregulation of coenzyme a transferase combined with alcohol-aldehyde dehydrogenase overexpression leads to predominantly alcohologenic Clostridium acetobutylicum fermentations. J Bacterial, 185, 3644-3653. 

Many strains of Clostridium botulinum synthesize an exoenzyme (C3) which catalyzes the specific ADP-ribosylation of a 21 kUodalton substrate (1). AH C3-producing strains also make either Cl or D neurotoxins. Botulinal neurotoxins block the release of neurotransmitters from nerve terminals and at the start of our work the Cl and D neurotoxins had been reported to be ADP-ribosyl transferases (2, 3). Hence we decided to determine if there was any relationship between C3-catalyzed ADP-ribosylation and nemosecretion. Here we show that when C3 is introduced into die cytosol of PC12 cells it induces a morphological differentiation but does not alter secretion of a neurotransmitter. By contrast the introduction of a neurotoxin into the cytosol of PC 12 cells does block secretion but causes no shape change. 

Cary, J.W, Petersen, D.J., Papoutsakis, E.T., and Bennett, G.N. (1990) Cloning and expression of Clostridium acetobutylicum ATCC 824 acetoacetyl-coenzyme Aacetate/butyrate coenzyme A-transferase in Escherichia coli. Appl Envirorc Microbiol, 56, 1576—1583. 

Chotani G, Dodge T, Hsu A, Kumar M, LaDuca R, Trimbur D, Weyler W, Sanford K (2000) The commercial production of chemicals using pathway engineering. Biochim Biophys Acta 1543 434-455 Clark SW, Bennett GN, Rudolph FB (1989) Isolation and characterization of mutants of Clostridium acetobutylicum ATCC 824 deficient in acetoacetyl-coenzyme A acetate/butyrate coenzyme A-transferase (EC 2.8.3.9) and in other solvent pathway enzymes. Appl Environ Microbiol 55 970-976 Cocks GT, Aguilar J, Lin EGG (1974) Evolution of L-1,2-propanediol catabolism in Escherichia coli by recruitment of enzymes for L-fucose and L-lactate metabolism. J Bacteriol 118 83-88... 

Colby GD (1993) CoA-transferase and 3-hydroxybutyryl-CoA Dehydrogenases acetoacetyl-CoA-reacting enzymes from Clostridium beijerinckii NRRL B593. PhD thesis, Virginia Polytechnic Institute and State University, Blacksburg VA... 

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