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1.
We studied 20 electrophoretic loci in two populations ofAteles (Ateles paniscus paniscus andAteles paniscus chamek). We observed intrapopulational variation at the following loci: esterase D, glyoxalase 1, adenosine deaminase (A. p. chamek) and carbonic anhydrase 2 (A. p. paniscus). The two populations share the most frequent alleles at 17 loci, but we noted great differences in glyoxalase 1, adenosine deaminase and phosphoglucomutase 1.A. p. paniscus is monomorphic for theGLO1 *1 allele, which has a frequency of 6% inA. p.chamek. They did not share alleles in relation to the ADA and PGM1 loci. We found a CA2 allele, named hereCA2 *1, which has not been described previously in other neotropical primates (Sampaio et al., 1991a), inA. p. paniscus. The present results suggest that the geographical isolation represented by the Rio Amazonas has lasted long enough to support this level of divergence. These observations taken together with chromosomal findings, led us to endorse the proposal of two distinct species:Ateles paniscus andAteles chamek.  相似文献   

2.
The present study was conducted to investigate the capability of Haloarcula marismortui to synthesize esterases and lipases, and the effect of physicochemical conditions on the growth and the production of esterases and lipases. Finally, the effect of NaCl concentration and temperature on esterase and lipase activities was studied using intracellular crude extracts. In order to confirm the genomic prediction about the esterase and lipase synthesis, H. marismortui was cultured on a rich medium and the crude extracts (intra- or extracellular) obtained were assayed for both activities using p-nitrophenyl esters and triacylglycerides as substrates. Studies on the kinetics of growth and production of esterase and lipase of H. marismortui were performed, reaching a maximum growth rate of 0.053 h−1 and maximal productions of intracellular esterase and lipase of 2.094 and 0.722 U l−1 using p-nitrophenyl valerate and p-nitrophenyl laurate, respectively. Both enzymes were produced as growth-associated metabolites. The effects of temperature, pH, and NaCl concentration on the growth rate and production of enzymes were studied by using a Box–Behnken response surface design. The three response variables were significantly influenced by the physicochemical factors and an interaction effect between temperature and NaCl concentration was also evidenced. The surface response method estimated the following maximal values for growth rate and productions of esterase and lipase: 0.086 h−1 (at 42.5°C, pH 7.4, and 3.6 mol l−1 NaCl), 2.3 U l−1 (at 50°C, pH 7.5, and 4.3 mol l−1 NaCl), and 0.58 U l−1 (at 50°C, pH 7.6, and 4.5 mol l−1 NaCl), respectively. Esterases were active at different salt concentrations, showing two optimal activities (at 0.5 and 5 mol l−1 NaCl), which suggested the presence of two different esterases. Interestingly, in the absence of salt, esterase retained 50% residual activity. Esterases and lipase activities were maximal at 45°C and inactive at 75°C. This study represents the first report evidencing the synthesis of esterase and lipase by H. marismortui.  相似文献   

3.
The genome sequence of Streptomyces coelicolor A3(2) contains 51 putative lipase and esterase genes mostly of unknown function. The gene estB (locus SCO 6966) was expressed as a His-tagged protein in E. coli. Esterase B was active at low temperatures exerting its maximum activity at 30°C and retaining more than 25% of its activity at 4°C. The optimum pH was 8–8.5. The enzyme was active against short synthetic p-nitrophenylesters (C2–C10) with maximum activity towards the acetate ester (C2). The esterase was tested on 13 series of racemic esters of potential interest for the synthesis of chiral pharmaceutical compounds. 4 of the series were substrates and a modest degree of enantioselectivity was observed (enantiomeric ratios of 1.1–1.9).  相似文献   

4.
A highly enantioselective l-menthyl acetate esterase was purified to homogeneity from Burkholderia cepacia ATCC 25416, with a recovery of 4.8% and a fold purification of 22.7. The molecular weight of the esterase was found to be 37 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The N-terminal amino acid sequence was “MGARTDA”, and there was no homology in contrast to other Burkholderia sp. esterases. This enzyme preferentially hydrolyzed short-chain fatty acid esters of menthol with high stereospecificity and high hydrolytic activity, while long-chain l-menthyl esters were poor substrates. Considered its substrate specificity and N-terminal sequence, this esterase was concluded as a new enzyme belonging to the carboxylesterase group (EC 3.1.1.1) of esterase family. The optimum temperature and pH for enzyme activity using racemic menthyl acetate as substrate were 30 °C and 7.0, respectively. The esterase was more stable in the pH range of 7.0–9.0 and temperature range of 30–40 °C. Hydrolytic activity was enhanced by Ca2+, K+ and Mg2+, but completely inhibited by Hg2+, Cu2+, ionic detergents and phenylmethylsulfonyl fluoride (PMSF) at 0.01 M concentration.  相似文献   

5.
The O-polysaccharide (O-antigen) of Escherichia coli O19ab was studied by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy. The following structure of the linear pentasaccharide repeating unit was established:→2)-α-l-Rhap-(1→2)-α-l-Rhap-(1→2)-α-l-Rhap-(1→2)-α-d-Glcp-(1→3)-α-d-GlcpNAc6Ac-(1→where the degree of O-acetylation of GlcNAc is ∼33%. The O-antigen gene cluster of E. coli O19ab was sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the E. coli O19ab-antigen structure.  相似文献   

6.
We identify an esterase isozyme inDrosophila melanogaster, EST 23, which shares biochemical, physiological, and genetic properties with esterase E3, which is involved in resistance to organophosphate insecticides inLucilia cuprina. Like E3, theD. melanogaster EST 23 is a membrane-bound -esterase which migrates slowly toward the anode at pH 6.8. Both enzymes have similar preferences for substrates with shorter acid side chain lengths. Furthermore, on the basis of their high sensitivity to inhibition by paraoxon and their insensitivity to inhibition by eserine sulfate, both enzymes were classified as subclass I carboxylesterases. The activity of each enzyme peaks early in development and, again, in the adult stage. Both enzymes are found in the male reproductive system and larval and adult digestive tissues, the latter being consistent with a role for these enzymes in organophosphate resistance. Fine structure deficiency mapping localizedEst 23 to cytological region 84D3 to E1-2 on the right arm of chromosome 3. Moreover, we show that the genes encoding three other esterase phenotypes also map to the same region; these phenotypes involve allozymic differences in EST 9 (formerly EST C), ali-esterase activity, defined by the hydrolysis of methyl butyrate, and malathion carboxylesterase activity, defined by hydrolysis of the organophosphate malathion. This cluster corresponds closely to that encompassing E3 and malathion carboxylesterase on chromosome 4 inL. cuprina, the homologue of chromosome 3R inD. melanogaster.  相似文献   

7.
A gene encoding an esterase (estO) was identified and sequenced from a gene library screen of the psychrotolerant bacterium Pseudoalteromonas arctica. Analysis of the 1,203 bp coding region revealed that the deduced peptide sequence is composed of 400 amino acids with a predicted molecular mass of 44.1 kDa. EstO contains a N-terminal esterase domain and an additional OsmC domain at the C-terminus (osmotically induced family of proteins). The highly conserved five-residue motif typical for all α/β hydrolases (G × S × G) was detected from position 104 to 108 together with a putative catalytic triad consisting of Ser106, Asp196, and His225. Sequence comparison showed that EstO exhibits 90% amino acid identity with hypothetical proteins containing similar esterase and OsmC domains but only around 10% identity to the amino acid sequences of known esterases. EstO variants with and without the OsmC domain were produced and purified as His-tag fusion proteins in E. coli. EstO displayed an optimum pH of 7.5 and optimum temperature of 25°C with more than 50% retained activity at the freezing point of water. The thermostability of EstO (50% activity after 5 h at 40°C) dramatically increased in the truncated variant (50% activity after 2.5 h at 90°C). Furthermore, the esterase displays broad substrate specificity for esters of short-chain fatty acids (C2–C8).  相似文献   

8.
A cDNA encoding a bifunctional acetylxylan esterase/xylanase, XynS20E, was cloned from the ruminal fungus Neocallimastix patriciarum. A putative conserved domain of carbohydrate esterase family 1 was observed at the N-terminus and a putative conserved domain of glycosyl hydrolase family 11 was detected at the C-terminus of XynS20E. To examine the enzyme activities, XynS20E was expressed in Escherichia coli as a recombinant His6 fusion protein and purified by immobilized metal ion-affinity chromatography. Response surface modeling combined with central composite design and regression analysis was then applied to determine the optimal temperature and pH conditions of the recombinant XynS20E. The optimal conditions for the highest xylanase activity of the recombinant XynS20E were observed at a temperature of 49°C and a pH of 5.8, while those for the highest carbohydrate esterase activity were observed at a temperature of 58°C and a pH of 8.2. Under the optimal conditions for the enzyme activity, the xylanase and acetylxylan esterase specific activities of the recombinant XynS20E toward birchwood xylan were 128.7 and 873.1 U mg−1, respectively. To our knowledge, this is the first report of a bifunctional xylanolytic enzyme with acetylxylan esterase and xylanase activities from rumen fungus.  相似文献   

9.
Kornyeyev  D.  Logan  B.A.  Holaday  A.S. 《Photosynthetica》2002,40(1):77-84
When dark-acclimated cotton (Gossypium hirsutum L. cv. Coker 312) leaves, pre-treated with lincomycin to inhibit chloroplast protein repair processes, were exposed to 10 °C and a PPFD of 500 mol m-2 s-1, the proportion of excitation energy entering photochemistry (P) increased, but only to 5 % of the total energy absorbed at steady state levels of P, which were reached at 40 min of irradiation. Thermal dissipation (D) of absorbed energy increased throughout the 360 min irradiation period and accounted for the greatest portion of absorbed energy at 10 °C. When D was partitioned into constitutive (DCON), regulated (DREG), and photoinhibitory (DPI) components, it was primarily composed of DREG, the readily reversible portion of D. However, the induction of D was slow at 10 °C. Sixty minutes were required for D to reach 70 % of the energy absorbed. Considerable absorption of energy in excess of that utilized in photochemistry or dissipated thermally (designated as E) occurred, especially during induction of P and D. Over the irradiation period, the time-dependent averaged E exhibited an inverse, linear relationship with the ratio of variable (Fv) to maximum (Fm) fluorescence (PS2 efficiency) and a linear relationship with DPI. We propose that time-dependent averaged E may be useful for estimating the potential for damage to PS2 under stressful environmental conditions.  相似文献   

10.
A thermostable esterase from the hyperthemophilic archaeonSulfolobus solfataricus was partially purified 590-fold with 16.2% recovery. The partially purified esterase had a specific activity of 29.5μmol min−1 mg−1 when the enzyme activity was determined usingp-nitrophenyl butyrate as a substrate. The apparent molecular weight was about 100 kDa, while the optimum temperature and pH for esterase were 75°C and 8.0, respectively. The enzyme showed high thermal stability and solvent tolerance in comparison to its mesophilic counterpart. The enzyme also showed chiral resolution activity for (S)-ibuprofen, indicating thatS. solfataricus esterase can be used for the production of commercially important chiral drugs.  相似文献   

11.
The degradation rate of the D1 polypeptide was measured in threeSynechocystis PCC 6803 mutantsin vivo. Mutations were introduced into a putative cleavage area of the D1 polypeptide (QEEET motif) and into the PEST-like area. PEST sequences are often found in proteins with a high turnover rate. The QEEET-motif mutants are CA1 [(E242-E244);Q241H] and E243K, and the third mutation, E229D, was directed to the PEST-like area. During high-light illumination (1500 mol photons m-2s-1) that induced photoinhibition of photosystem II (PSII), the half-life time of the D1 polypeptide in mutant E229D (t 1/2=35 min) was about twice as long as in AR (control strain) cells (t 1/2=19 min). In growth light (40 mol photons m-2s-1), the degradation rate of the D1 polypeptide in E229D and AR strains was the same (t 1/25 h). In growth light the D1 polypeptide was degraded faster in both QEEET-motif mutants than in the AR strain, but in photoinhibitory light the degradation rates were similar. According to these results, the highly conservative QEEET motif as such is not required for the proteolytic cut of the D1 polypeptide, but it does affect the rate of degradation. No simple correlation existed between the degradation rate of the D1 polypeptide and the susceptibility of PSII to photoinhibition in mutant and AR cells under our experimental conditions.  相似文献   

12.
Zhang J  Liu J  Zhou J  Ren Y  Dai X  Xiang H 《Biotechnology letters》2003,25(17):1463-1467
The lipA gene encoding a thermostable esterase was cloned from Thermoanaerobacter tengcongensis and overexpressed in Escherichia coli. The recombinant esterase, with a molecular mass of approx. 43 kDa determined by SDS-PAGE, was purified to homogeneity through Sephadex G-100 gel filtration. The purified enzyme actively hydrolyzed tributyrin but not olive oil. Maximum activity was observed on p-nitrophenyl (NP)-propionate (C3) and p-NP-butyrate (C4), with little activity towards p-NP-palmitate (C16). The esterase was optimally active at 70 °C (over 15 min) and at pH 9. It is highly thermostable, with a residual activity greater than 80% after incubation at 50 °C for more than 10 h. The activity was not inhibited by 5 mM EDTA and PMSF, indicating the esterase is not a metalloenzyme and may contain a specific structure around the catalytic serine residue. In addition, it was stable for 1 h at 37 °C in 1% CHAPS and Triton X-100 but not stable in 1% Tween 20 or SDS.  相似文献   

13.
The purpose of our work was to observe the influence of probiotic preparation BioPlus 2B on average daily gains of weaned pigs, feed efficiency and to evaluate some metabolic indices. The weaned pigs, at the age of 42 days, were included into the trial and divided into four groups. Pigs in groups A (n = 5) and B (n = 4) received BioPlus 2B also before weaning. Only group A received BioPlus 2B, at the concentration of 3.2 × 109 CFU per kg of feed, after weaning continually. Groups C (n = 5) and D (n = 4) did not receive BioPlus 2B until the start of the trial, but group C was given BioPlus 2B at the same concentration as group A during the experiment. Blood samples for determination of metabolic indices were collected at the start of experiment, i.e. on 42nd day of pigs life, and then on 56th, 70th, 84th and 91st days of pigs life. The following biochemical indices were evaluated within the trial: serum levels of total proteins, albumin, urea, total cholesterol and total lipids. Total serum protein level (p < 0.0001) and serum albumin level (p = 0.0024) in groups A and B were significantly higher in comparison with groups C and D on day 56 of pigs life. Serum urea level in group D was significantly (p = 0.049) higher than in group A on 70th day of pigs life. Serum level of total cholesterol in group B on day 56 and 84 of pigs life was significantly (p = 0.0004) higher than in groups C and D. Total serum lipid level was significantly (p < 0.05) higher in B group compared to other groups on 56th, 70th, 84th days of pigs life. Average daily gains (ADG) in A group, even if non-significantly, were about 10% better than in groups B, C, D between 57th and 77th days of pigs life. ADG in groups A and B were 14% better in comparison with that in groups C and D between 78th and 91st days of pigs life (p = 0.036). The best feed efficiency in the trial was in group A, approximately 13%, 16% and 21% better than that in the groups B, C and D, repectively. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.  相似文献   

14.
Isoelectric focusing was used to identify five alleles at the locus determining the production of the sixth component of complement (C6) in the dog. Four of these alleles,C6 1, C62, C64,andC6 5,were studied in family pedigrees and shown to be inherited in a codominant autosomal fashion. All alleles except forC6 4occurred commonly in the multiple breeds tested. This investigation was supported by Grant HL 17265 awarded by the National Institute of Heart, Lung, and Blood Diseases, DHEW, and by Grants CA 18105 and CA 31787 awarded by the National Cancer Institute.  相似文献   

15.
A novel esterase gene (e25) was identified from Altererythrobacter epoxidivorans CGMCC 1.7731T by genome sequence screening. The e25 gene is 948 nucleotides in length and encodes a 315?amino acid protein (E25) with a predicted molecular mass of 33,683 Da. A phylogenetic tree revealed that E25 belongs to the hormone-sensitive lipase (HSL) family of lipolytic enzymes. An activity assay of E25 showed that it exhibited the highest catalytic efficiency when using p-nitrophenyl caproate (C6) as a substrate. The optimum pH and temperature were determined to be approximately pH 9 and 45°C, and the Km and Vmax values were 0.12?mM and 1,772?µmol/min/mg, respectively. After an incubation at 40°C for 80?min, E25 retained 75% of its basal activity. The enzyme exhibited good tolerance to metal cations, such as Ba2+, Ca2+, and Cu2+ (10?mM), but its activity was strongly inhibited by Co2+, Ni2+, Mn2+, and Zn2+. The E25 enzyme was stimulated by glycerol and retained over 60% of its basal activity in the presence of 1% Tween-80 and Triton X-100. Overall, the activity of E25 under alkaline conditions and its organic solvent and detergent tolerance indicate that E25 could be useful as a novel industrial catalyst in biotechnological applications.  相似文献   

16.
Summary Escherichia coli Rl is an Ag+-resistant strain that, as we have shown recently, harbours at least two large plasmids, pJT1 (83 kb) and pJT2 (77 kb). Tn5-Mob was introduced into theE. coli Rl host replicon via conjugation on membrane filters. The transfer functions of plasmid RP4-4 were provided in this process and Tn5-Mob clones mated withE. coli C600 yielded Ag+-resistant transconjugants. This mobilization procedure allowed transfer and expression of pJT1 Ag+ resistance inE. coli C600. Prior to use of Tn5-Mob mobilization, it was not possible to transfer Ag+-resistant determinant(s) intoE. coli by conjugation or transformation including high-voltage electroporation.E. coli C600 containing PJTI and PJT2 displayed decreased accumulation of Ag+ similar toE. coli R1.E. coli C600 could not tolerate 0.1 and 0.5 mM Ag+, rapidly accumulated Ag+ and became non-viable. Tn5-Mob mobilization may be useful in the study of metal resistance in bacteria, especially in strains not studied for resistance mechanisms.  相似文献   

17.
Walter K. Dodds 《Oecologia》1991,85(4):572-580
Summary Interactions between epiphytes, epiphyte grazers and the filamentous green alga Cladophora glomerata (L.) Kuetzing were explored with smaples from rivers in Montana. Extracts of C. glomerata lowered photosynthetic rates of Nitzschia fonticola Grunow (an epiphytic diatom). Nutrient enrichment showed that C. glomerata from the Madison River was N deficient and its epiphytes were P deficient on 2 dates and N deficient on one date, while no nutrient deficiencies were detected in samples from 3 other rivers; this implies there was little nutrient competition between the epiphytes and C. glomerata. Epiphytes lowered drag on C. glomerata tufts and current velocity inside the tufts, apparently by decreasing the effective surface area. Lower drag may decrease detachment, but lowering current velocity from 8 to 0 cm s-1 resulted in a 100 % decrease in photosynthesis. Light absorption by epiphyte pigments may lower photosynthetic rate of C. glomerata when irradiance is below 200–500 E m-2 s-1, and protect against photoinhibition above this irradiance range. Invertebrate grazers (predominantly Baetis tricaudatus Dodds, Trycorythodes minutus Traver and Brachycentrus occidentalis Banks) at high densities removed 75% of epiphytes and B. occidentalis grazed on C. glomerata. Invertebrates regenerated a mean of 0.16 mol NH inf4 sup+ individual-1 d-1 which could have enhanced growth of downstream C. glomerata. Competition and grazing were not the only interactions in the C. glomerata community, positive (mutualistic) interactions were also important.  相似文献   

18.
Four mutations in the mitochondrial cytochromeb ofS. cerevisiae have been characterized with respect to growth capacities, catalytic properties, ATP/2e ratio, and transmembrane potential. The respiratory-deficient mutant G137E and the three pseudo-wild type revertants E137 + I147F, E137 + C133S, and E137 + N256K were described previously (Tron and Lemesle-Meunier, 1990; Di Ragoet al., 1990a). The mutant G137E is unable to grow on respiratory substrates but its electron transfer activity is partly conserved and totally inhibited by antimycin A. The secondary mutations restore the respiratory growth at variable degree, with a phosphorylation efficiency of 12–42% as regards the parental wild type strain, and result in a slight increase in the various electron transfer activities at the level of the whole respiratory chain. The catalytic efficiency for ubiquinol was slightly (G137E) or not affected (E137 + I147F, E137 + C133S, and E137 + N256K) in these mutants. Mutation G137E induces a decrease in the ATP/2e ratio (50% of the W.T. value) and transmembrane potential (60% of the W.T. value) at thebc1 level, whereas the energetic capacity of the cytochrome oxidase is conserved. Secondary mutations I147F, C133S, and N256K partly restore the ATP/ 2e ratio and the transmembrane potential at thebc1 complex level. The results suggest that a partial decoupling of thebc1 complex is induced by the cytochromeb point mutation G137E. In the framework of the protonmotive Q cycle, this decoupling can be explained by the existence of a proton wire connecting centers P and N in the wild typebc1 complex which may be amplified or uncovered by the G137E mutation when the bc1 complex is functioning.  相似文献   

19.
The wild‐type HIV‐1 capsid protein (CA) self‐assembles in vitro into tubular structures at high ionic strength. We report solid state nuclear magnetic resonance (NMR) and electron microscopy measurements on these tubular CA assemblies, which are believed to contain a triangular lattice of hexameric CA proteins that is similar or identical to the lattice of capsids in intact HIV‐1. Mass‐per‐length values of CA assemblies determined by dark‐field transmission electron microscopy indicate a variety of structures, ranging from single‐wall tubes to multiwall tubes that approximate solid rods. Two‐dimensional (2D) solid state 13C? 13C and 15N? 13C NMR spectra of uniformly 15N,13C‐labeled CA assemblies are highly congested, as expected for a 25.6 kDa protein in which nearly the entire amino acid sequence is immobilized. Solid state NMR spectra of partially labeled CA assemblies, expressed in 1,3‐13C2‐glycerol medium, are better resolved, allowing the identification of individual signals with line widths below 1 ppm. Comparison of crosspeak patterns in the experimental 2D spectra with simulated patterns based on solution NMR chemical shifts of the individual N‐terminal (NTD) and C‐terminal (CTD) domains indicates that NTD and CTD retain their individual structures upon self‐assembly of full‐length CA into tubes. 2D 1H‐13C NMR spectra of CA assemblies recorded under solution NMR conditions show relatively few signals, primarily from segments that link the α‐helices of NTD and CTD and from the N‐ and C‐terminal ends. Taken together, the data support the idea that CA assemblies contain a highly ordered 2D protein lattice in which the NTD and CTD structures are retained and largely immobilized.  相似文献   

20.
A new inherited variant of carbonic anhydrase I (CA I), designated CA INagasaki 1 (CA INGS 1), was discovered during a survey of hemolysates from 5852 individuals from the cities of Hiroshima and Nagasaki in Japan. Analysis of the amino acid composition of a tryptic peptide from the CA INGS 1 variant indicated that a glutaminyl residue was substituted for an arginyl residue at position 76. Heat degradation studies showed that the CA INGS 1 variant was less stable than normal CA I. The CO2 hydrase and esterase activities of the normal and variant carbonic anhydrases I, as well as the relative amounts of the two enzymes in heterozygotes, were similar.This work was supported in part by Contract E(11-1)-1552 with the Energy Research and Development Administration, Washington, D.C. (to J. V. Neel), and by U.S. Public Health Service Grant GM-24681.  相似文献   

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