Purification and characterization of a long-chain acyl coenzyme A thioesterase from Rhodopseudomonas sphaeroides

Boyce, Stephen G.; Leuking, Donald R.

doi:10.1021/bi00296a023

Cited by 16 publications

(15 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, both PhaG and thioesterase II are essential for extracellular production of 3HD. Surprisingly, two thioesterases whose activities are similar to those of E. coli were found in Rhodopseudomonas sphaeroides (4). Similar thioesterase complementation occurring in dissimilar bacteria suggests that these enzymes play important roles (22).…”

Section: Discussionmentioning

confidence: 92%

Thioesterase II of Escherichia coli Plays an Important Role in 3-Hydroxydecanoic Acid Production

Zheng

Gong

Liu

et al. 2004

Appl Environ Microbiol

View full text Add to dashboard Cite

3-Hydroxydecanoic acid (3HD) was produced in Escherichia coli by mobilizing (R)-3-hydroxydecanoyl-acyl carrier protein-coenzyme A transacylase (PhaG, encoded by the phaG gene). By employing an isogenic tesB (encoding thioesterase II)-negative knockout E. coli strain, CH01, it was found that the expressions of tesB and phaG can up-regulate each other. In addition, 3HD was synthesized from glucose or fructose by recombinant E. coli harboring phaG and tesB. This study supports the hypothesis that the physiological role of thioesterase II in E. coli is to prevent the abnormal accumulation of intracellular acyl-coenzyme A.

show abstract

Section: Discussionmentioning

confidence: 92%

Thioesterase II of Escherichia coli Plays an Important Role in 3-Hydroxydecanoic Acid Production

Zheng

Gong

Liu

et al. 2004

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…We are currently attempting to isolate mutants deficient in both thioesterase activities. It should be noted that another gram-negative bacterium, Rhodopseudomonas sphaeroides has two thioesterase activities very similar to those of E. coli (8). The finding that two such dissimilar bacteria possess similar thioesterase complements suggests that these enzymes play an important but as yet not understood role.…”

Section: Discussionmentioning

confidence: 99%

“…Long-chain acyl thioesterases occur in a wide variety of organisms, and the properties of several of these proteins have been investigated (8,11,19,23,28). The physiological role of these enzymes is, however, still unclear.…”

mentioning

confidence: 99%

Genetic and biochemical characterization of an Escherichia coli K-12 mutant deficient in acyl-coenzyme A thioesterase II

Narasimhan¹,

Lampi²,

Cronan³

1986

J Bacteriol

View full text Add to dashboard Cite

Mutants of Escherichia coli deficient in thioesterase II activity were isolated by taking advantage of the fact that thioesterase I specifically hydrolyzes long-chain (C12 to C18) acyl coenzyme A (CoA) esters but is unable to cleave the short-chain substrate decanoyl-CoA. One of these lesions (designated tesBi) reduces thioesterase II activity to about 10% of the normal level. The mutant enzyme activity was abnormally labile to temperature, but It was normal in all the other characteristics examined (pH optimum, Km for decanoyl-CoA, molecular weight). The level of thioesterase I activity was unaffected by the tesBI lesion. The tesB locus was mapped with a closely linked Tn1O insertion. tesB was mapped to minute 10 of the E. coli linkage map, close to the lon locus. The clockwise gene order is Ion tesB acrA dnaZ. The tesB mutation is recessive. We found no phenotype for the mutation. The fatty acid compositions of the phospholipids, lipid A, and lipoprotein components are normal in thioesterase II mutants. These data show that thioesterases I and II of E. coli are encoded by different genetic loci and strongly suggest that tesB is the structural gene for thioesterase II.

show abstract

“…A histidine residue present at position 58 in thioesterase II has been implicated in the cleavage of the thioester bond [10,11]. Two thioesterases with striking similarities in their physical properties to thioesterase I and II have been reported from photosynthetic bacteria, Rhodopseudomonas sphaeroides [12,13].…”

mentioning

confidence: 99%

Characterization of a novel long‐chain acyl‐CoA thioesterase from Alcaligenes faecalis

et al. 2006

View full text Add to dashboard Cite

Long-chain acyl-CoA thioesterases (EC 3.1.2.2) hydrolyze acyl-CoA esters to nonesterified fatty acids and coenzyme A (CoASH) [1]. These are ubiquitously expressed in bacteria, yeast, plants and mammals, and in most cell compartments, such as endoplasmic reticulum, cytosol, mitochondria and peroxisomes. Several unrelated thioesterases have been purified to homogeneity from plants, animals, and bacteria, and the cDNAs encoding several of them have been cloned and sequenced [2][3][4][5][6][7]. Although the physiological functions of these enzymes remain largely unknown, it is speculated that they regulate lipid metabolism by maintaining appropriate concentrations of acyl-CoA, CoASH, and nonesterified fatty acids. The only established function for acyl-CoA thioesterases is in the termination of fatty acid synthesis in eukaryotes [8].Two thioesterases, I and II, that cleave acyl-CoA molecules in vitro have been characterized from A novel long-chain acyl-CoA thioesterase from Alcaligenes faecalis has been isolated and characterized. The protein was extracted from the cells with 1 m NaCl, which required 1.5-fold, single-step purification to yield near-homogeneous preparations. In solution, the protein exists as homomeric aggregates, of mean diameter 21.6 nm, consisting of 22-kDa subunits. MS ⁄ MS data for peptides obtained by trypsin digestion of the thiosterase did not match any peptide from Escherichia coli thioesterases or any other thioesterases in the database. The thioesterase was associated exclusively with the surface of cells as revealed by ultrastructural studies using electron microscopy and immunogold labeling. It hydrolyzed saturated and unsaturated fatty acyl-CoAs of C 12 to C 18 chain length with V max and K m of 3.58-9.73 lmolAEmin )1 AE(mg protein) )1 and 2.66-4.11 lm, respectively. A catalytically important histidine residue is implicated in the active site of the enzyme. The thioesterase was active and stable over a wide range of temperature and pH. Maximum activity was observed at 65°C and pH 10.5, and varied between 60% and 80% at temperatures of 25-70°C and pH 6.5-10. The thioesterase also hydrolyzed p-nitrophenyl esters of C 2 to C 12 chain length, but substrate competition experiments demonstrated that the long-chain acyl-CoAs are better substrates for thioesterase than p-nitrophenyl esters. When assayed at 37 and 20°C, the affinity and catalytic efficiency of the thioesterase for palmitoleoyl-CoA and cis-vaccenoyl-CoA were reduced approximately twofold at the lower temperature, but remained largely unaltered for palmitoyl-CoA.Abbreviations DTNB, 5,5¢-dithiobis(2-nitrobenzoic acid); TEM, transmission electron microscopy.

show abstract

Purification and characterization of a long-chain acyl coenzyme A thioesterase from Rhodopseudomonas sphaeroides

Cited by 16 publications

References 42 publications

Thioesterase II of Escherichia coli Plays an Important Role in 3-Hydroxydecanoic Acid Production

Thioesterase II of Escherichia coli Plays an Important Role in 3-Hydroxydecanoic Acid Production

Genetic and biochemical characterization of an Escherichia coli K-12 mutant deficient in acyl-coenzyme A thioesterase II

Characterization of a novel long‐chain acyl‐CoA thioesterase from Alcaligenes faecalis

Contact Info

Product

Resources

About