Clostridium kluyveri

PHAs consisting of 4HB are other examples which can be synthesized from simple, unrelated carbon sources. Recombinant strains of E. coli expressing phaCRe and orfZ from Clostridium kluyveri synthesized poly(3HB-co-4HB) or even poly(4HB) homopolyester if 4-hydroxybutyrate was used as carbon source [90]. When additional succinate degradation genes from C. kluyveri were expressed in the recombinant E. coli, poly(3HB-co-4HB) with a low content of 4HB was synthesized from glucose [118]. 

Lynen had studied chemistry in Munich under Wieland his skill as a chemist led to the successful synthesis of a number of fatty acyl CoA derivatives which proved to be substrates in the catabolic pathway. Many of these C=0 or C=C compounds had characteristic UV absorption spectra so that enzyme reactions utilizing them could be followed spectrophotometrically. This technique was also used to identify and monitor the flavoprotein and pyridine nucleotide-dependent steps. Independent evidence for the pathway was provided by Barker, Stadtman and their colleagues using Clostridium kluyveri. Once the outline of the degradation had been proposed the individual steps of the reactions were analyzed very rapidly by Lynen, Green, and Ochoa s groups using in the main acetone-dried powders from mitochondria, which, when extracted with dilute salt solutions, contained all the enzymes of the fatty acid oxidation system. 

Hashimoto, H., Gtlnther, H., Simon, H. The stereochemistry of vinylacetyl-CoA-isomerase of Clostridium kluyveri. FEBS Letters 33, 81—83 (1973). 

The biochemical reduction of a, -unsaturated -haloaliphatic acids by means of Clostridium kluyveri yielded halogen-free satmated acids. The same products were obtained from saturated -halo acids. However, the same microorganism converted ix, -unsaturated a-halo acids to saturated a-halo acids with R configuration. Yields of reduction of a-fluoro-, a-chloro- and a-bromocrotonic acid ranged from 30% to 100% [330]. 

Selected entries from Methods in Enzymology [vol, page(s)] Determination of FMN and FAD by fluorescence titration with apoflavodoxin, 66, 217 purification of flavin-adenine dinucleotide and coenzyme A on p-acetoxymercurianiline-agarose, 66, 221 a convenient biosynthetic method for the preparation of radioactive flavin nucleotides using Clostridium kluyveri, 66, 227 isolation, chemical synthesis, and properties of roseoflavin, 66, 235 isolation, synthesis, and properties of 8-hydroxyflavins, 66, 241 structure, properties and determination of covalently bound flavins, 66, 253 a two-step chemical synthesis of lumiflavin, 66, 265 syntheses of 5-deazaflavins, 66, 267 preparation, characterization, and coenzymic properties of 5-carba-5-deaza and 1-... 

A thiolase (Eq. 13-35) from Clostridium kluyveri is one of only two known selenoproteins that contain selenomethionine.569 However, the selenomethionine is incorporated randomly in place of methionine. This occurs in all proteins of all organisms to some extent and the toxicity of selenium may result in part from excessive incorporation of selenomethionine into various proteins. 

The most obvious route of metabolism of propionyl-CoA is further (1 oxidation which leads to 3-hydroxypropionyl-CoA (Fig. 17-3, step a). This appears to be the major pathway in green plants.17 Continuation of the (1 oxidation via steps a-c of Fig. 17-3 produces the CoA derivative of malonic semialdehyde. The latter can, in turn, be oxidized to malonyl-CoA, a P-oxoacid which can be decarboxylated to acetyl-CoA. The necessary enzymes have been found in Clostridium kluyveri,70 but the pathway appears to be little used. 

The fermentation of Eq. 17-31 is catalyzed by Clostridium kluyveri. The value of -AG is one of the lowest that we have considered but is still enough to provide easily for the synthesis of one molecule of ATP. 

Fredericks, W. W. and E. R. Stadtman The role of ferredoxin in the hydro-genase system from Clostridium kluyveri. J. Biol. Chem. 240, 4065—4071 (1965). 

Stadtman, E. R. and H. A. Barker Fatty acid synthesis by enzyme preparations of Clostridium kluyveri. VI. Reactions of acyl phosphates. J. Biol. Chem. 184, 769-793 (1950). 

Stern, J. R. Role of cofactors in pyruvate oxidation and synthesis by extracts of Clostridium kluyveri. In Non-Heme Iron Proteins Role in Energy Conversion, A. San Pietro, ed., Antioch Press, Yellow Springs, Ohio, pp. 199—210 (1965). 

Tomlinson, N. Carbon dioxide and acetate utilization by Clostridium kluyveri. II. Synthesis of amino acids. J. Biol. Chem. 209, 597—603 (1954). 

Bechamp (15) in 1867 was the first to describe methane production from a simple fermentation product, ethanol, and to attribute it to a microbial fermentation. The second product he found to be formed from ethanol was caproate, now known to be produced by Clostridium kluyveri (4). Thus, carbon-carbon bonds are not only destroyed early in the fermentation chain, they can also be reductively synthesized if the reaction is paired with the energy yielding oxidation of another substrate (c/. initial fermentations of acetate and ethanol and also of H2, Figure 1). Several successive fermentations may then be required to convert these products to methane and CO2. 

No NADP-specific activity has been detected in this organism nor in Rhodospirillum rubrum (37), but Clostridium kluyveri may possess both NAD and NADP linked GDH s (40). 

Simon s Clostridium kluyveri system has been used to reduce trans-cinnamic acid 309 to (2S, 3R)-[2,3- H2]-3-phenylpropionic acid 310, and this was converted by bromination and ammonolysis to (2RS,3S)-[3- Hi]phenyl-alanine 297, Hp = H (311), as in Scheme 82. A Japanese group recently reduced the diketopiperazine 311 at the exomethylene group with asymmetric induction and obtained (2R, 3R)-[2,3- H2]phenylalanine (312). 

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