scholarly journals Purification, Characterization, Molecular Cloning, and Expression of a New Aminoacylase fromStreptomyces mobaraensisThat Can HydrolyzeN-(Middle/Long)-chain-fatty-acyl-L-amino Acids as Well asN-Short-chain-acyl-L-amino acids

2009 ◽  
Vol 73 (9) ◽  
pp. 1940-1947 ◽  
Author(s):  
Mayuko KOREISHI ◽  
Yasuyuki NAKATANI ◽  
Manami OOI ◽  
Hiroyuki IMANAKA ◽  
Koreyoshi IMAMURA ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Cloning ◽  
Fatty Acyl ◽  
Cloning And Expression ◽  
Short Chain ◽  
Chain Acyl ◽  
Long Chain

Molecular Cloning and Expression of cDNAs Encoding Rat Brain and Liver Cytosolic Long-Chain Acyl-CoA Hydrolases

1997 ◽  
Vol 232 (1) ◽  
pp. 198-203 ◽  
Author(s):  
Junji Yamada ◽  
Takao Furihata ◽  
Noriko Iida ◽  
Takafumi Watanabe ◽  
Masakiyo Hosokawa ◽  
...  
Keyword(s):  
Rat Brain ◽  
Molecular Cloning ◽  
Cloning And Expression ◽  
Rat Brain And Liver ◽  
Chain Acyl ◽  
Long Chain

scholarly journals Higher-plant medium- and short-chain acyl-CoA oxidases: identification, purification and characterization of two novel enzymes of eukaryotic peroxisomal β-oxidation

1996 ◽  
Vol 320 (2) ◽  
pp. 607-614 ◽  
Author(s):  
Mark A. HOOKS ◽  
Kornelia BODE ◽  
Ivan COUÉE
Keyword(s):  
Molecular Mass ◽  
Subcellular Location ◽  
Maximal Activity ◽  
Fatty Acyl ◽  
Short Chain ◽  
Higher Plant ◽  
Medium Chain ◽  
Chain Acyl ◽  
Acyl Coa Oxidase ◽  
Long Chain

Medium- and short-chain acyl-CoA oxidases were identified in and subsequently purified from dark-grown maize plantlets. The oxidase showing preference for medium-chain fatty acyl-CoAs (C10–C14) was purified to homogeneity. The oxidase showing preference for short-chain fatty acyl-CoAs (C4–C8) was purified over 150-fold. Various catalytic properties confirmed these enzymes to be true acyl-CoA oxidases. They produced trans-2-enoyl-CoA and H2O2 from the saturated acyl-CoA, as verified by various independent assay techniques. They also exhibited FAD-dependent activity; i.e. removal of loosely bound FAD by gel filtration markedly reduced activity, which could be restored upon re-addition of FAD. They showed apparent Km values between 2 and 10 µM for the acyl-CoA substrate giving maximal activity, no activity with the corresponding free fatty acid, high pH optima (8.3–8.6) and a peroxisomal subcellular location. The medium-chain acyl-CoA oxidase was determined to be a monomeric protein with a molecular mass of 62 kDa. The short-chain acyl-CoA oxidase was shown to have a native molecular mass of 60 kDa, but exhibited a labile multimeric structure, as indicated by the elution of multiple peaks of activity during several chromatographic steps, and ultimately by the purification of a subunit of molecular mass 15 kDa. The medium- and short-chain acyl-CoA oxidases were demonstrated to be distinct from the maize equivalent of the cucumber glyoxysomal long-chain acyl-CoA oxidase previously purified and characterized [Kirsch, Loffler and Kindl (1986) J. Biol. Chem. 261, 8570–8575]. The maize long-chain acyl-CoA oxidase was partially purified to permit determination of its substrate specificity; it showed activity with a broad range of acyl-CoAs of chain length greater than C8, and maximal activity with C16. The implications of the existence of multiple acyl-CoA oxidases in the regulation of plant peroxisomal β-oxidation are discussed.

scholarly journals Effect of long-chain fatty acyl-CoA on mitochondrial and cytosolic ATP/ADP ratios in the intact liver cell

1984 ◽  
Vol 220 (2) ◽  
pp. 371-376 ◽  
Author(s):  
S Soboll ◽  
H J Seitz ◽  
H Sies ◽  
B Ziegler ◽  
R Scholz
Keyword(s):  
Liver Cell ◽  
Adenine Nucleotide ◽  
Intact Cell ◽  
Inhibitory Effect ◽  
Fatty Acyl ◽  
Fed State ◽  
Physiological Relevance ◽  
Chain Acyl ◽  
Long Chain

The effect of long-chain acyl-CoA on subcellular adenine nucleotide systems was studied in the intact liver cell. Long-chain acyl-CoA content was varied by varying the nutritional state (fed and starved states) or by addition of oleate. Starvation led to an increase in the mitochondrial and a decrease in the cytosolic ATP/ADP ratio in liver both in vivo and in the isolated perfused organ as compared with the fed state. The changes were reversed on re-feeding glucose in liver in vivo or on infusion of substrates (glucose, glycerol) in the perfused liver, respectively. Similar changes in mitochondrial and cytosolic ATP/ADP ratios occurred on addition of oleate, but, importantly, not with a short-chain fatty acid such as octanoate. It is concluded that long-chain acyl-CoA exerts an inhibitory effect on mitochondrial adenine nucleotide translocation in the intact cell, as was previously postulated in the literature from data obtained with isolated mitochondria. The physiological relevance with respect to pyruvate metabolism, i.e. regulation of pyruvate carboxylase and pyruvate dehydrogenase by the mitochondrial ATP/ADP ratio, is discussed.

scholarly journals A fixation procedure suitable for autoradiography of endogenous long-chain acyl carnitine.

1985 ◽  
Vol 33 (8) ◽  
pp. 744-748 ◽  
Author(s):  
M T Knabb ◽  
G G Ahumada ◽  
B E Sobel ◽  
J E Saffitz
Keyword(s):  
Subcellular Localization ◽  
Selective Extraction ◽  
Total Radioactivity ◽  
Water Soluble ◽  
Free Carnitine ◽  
Neonatal Rat ◽  
Short Chain ◽  
Tissue Processing ◽  
Chain Acyl ◽  
Long Chain

A tissue processing procedure was evaluated for fixation of endogenous long-chain acyl carnitine (LCA) to facilitate autoradiographic subcellular localization of this amphiphile. Suspensions of neonatal rat myocytes labeled with exogenous 14C-palmitoyl carnitine retained 85.2% of the radiolabel after tissue processing. Autoradiography demonstrated no significant translocation of radiolabeled LCA from myocytes to unlabeled sheep erythrocytes mixed in equal proportions and processed together. To evaluate endogenous LCA fixation, cultured myocytes were incubated for 3 days with 3H-carnitine. Radioactivity was distributed in LCA, short-chain acyl carnitine, and free carnitine pools in proportion to the physiological concentrations of the metabolites traced. Before tissue processing, LCA contained 4.5% of total radioactivity. After tissue processing, labeled water-soluble components were lost and 88% of the retained radioactivity was in the LCA pool. The enrichment of endogenous LCA radioactivity was attributable to the selective extraction of endogenous short-chain and free carnitine. Nearly 75% of endogenous LCA was preserved. In contrast, 99.5% of both endogenous short-chain and free carnitine were extracted. Thus, endogenous LCA can be selectively preserved, permitting quantitative subcellular localization of this amphiphile with ultrastructural autoradiography.

Molecular cloning and expression of key enzymes for biosynthesis of cysteine and related secondary non-protein amino acids

1994 ◽  
Vol 38 (2-3) ◽  
pp. 153-158 ◽  
Author(s):  
Kazuki Saito ◽  
Naoko Miura ◽  
Mami Yamazaki ◽  
Kazuyo Tatsuguchi ◽  
Makoto Kurosawa ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Cloning ◽  
Cloning And Expression ◽  
Key Enzymes ◽  
Protein Amino Acids
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