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1.
R.J. Pryce 《Phytochemistry》1973,12(3):507-514
A qualitative and quantitative analysis of the decomposition of unbuffered and buffered (pH 3–8) aqueous solutions of gibberellic acid (GA3) on autoclaving is recorded. The identified products, which vary in composition with pH, are iso-GA3 (II), iso-GA3 hydroxy acid (III), gibberellenic acid (IV), allogibberic acid (V), epiallogibbe detected after autoclaving in all cases. The identified products, in all cases, account for not less than 95% of the decomposition product, Dehydroallogibberic acid has not previously been recorded as an aqueous decomposition product of GA3 and its biological activity in the lettuce hypocotyl test is recorded. 相似文献
2.
A new natural product, 2(S),3(S)-3-hydroxy-4-methyleneglutamic acid (G3) has been isolated from seeds of Gleditsia caspica. The structure has been established by chemical and spectroscopic methods. Catalytic reduction of G3 yields 2(S),4(S)-4-methylglutamic acid and a new amino acid, 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid. Ozonolysis of G3 followed by oxidation gives 2(S),3(R)-3-hydroxyaspartic acid. The S- (or l-) configurations at C2 in G3 and in 2(S),3(S),4(S)-3-hydroxy-4-methyglutamic acid and the S-configurations at C3 for G3 and 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid and at C4 for 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid are inferred from the configurations at C2 in 2(S),4(S)-4-methylglutamic acid and at C2 and C3 in 2(S),3(R)-3-hydroxyaspartic acid. The seeds also contain appreciable quantities of 2(S),3(S),4(R)-3-hydroxy-4-methylglutami c acid (G1) and 2(S),4(R)-4-methylglutamic acid. 相似文献
3.
Rajasekhar Urla P. Pavan Kumar Prabhakar Sripadi Venkateswara Rao Khareedu Dashavantha Reddy Vudem 《Plant biotechnology reports》2017,11(5):259-270
Flax contains very high levels of alpha-linolenic acid (57%) and a fatty acid desaturase 3-coding sequence (Lufad3) of flax has been amplified from the RNA isolated from developing seeds. The deduced amino acid sequence of LuFAD3 showed the presence of three histidine motifs, six membrane spanning domains and an endoplasmic reticulum (ER) retention signal KSK, indicating its plausible localization into the ER. Flax is not amenable for genetic transformation and not suitable for functional validation of Lufad3 gene. Hence, rice with well-developed genetic transformation has been selected as heterologous host system. Coding sequence of Lufad3 driven by maize Ubi1 promoter has been introduced into indica rice by Agrobacterium tumefaciens-mediated genetic transformation. Southern analysis of putative transformants (T 0) revealed signals at variable lengths at >5 kbp indicating random integration of transgene into the genomes of different transformants. The Mendelian segregation observed for selectable marker gene hyg in both T1 and T2 generations confirmed stable inheritance and single-site integration of transgenes. As compared to untransformed control (UC), homozygous transgenic rice expressing Lufad3 showed higher levels of essential α-linolenic acid in leaves and seeds validating its functionality. 相似文献
4.
Hiroyuki Yamamoto Tadao Hayakawa 《International journal of biological macromolecules》1982,4(2):116-120
The oxidation of l-ascorbic acid and d-isoascorbic acid by the poly(l-lysine) (PLL), poly(d-lysine) (PDL), poly(l-ornithine), poly(l-arginine) and poly(l-histidne) Cu(II) complexes in phosphate buffer has been investigated. Most polypeptide-Cu(II) complexes showed an inhibitory effect. However, a comparison of the initial velocities of the enantiomeric polypeptide Cu(II) complexes gave the stereoselectivity indices (VO)PDL cu/(VO)PLL-Cu = 2.8 on l-ascorbic acid and 1.8 on d-isoascorbic acid at pH 5. At alkaline pH, the reverse was found for the stereoselective oxidation. At pH 10.5, the indices were calculated to be 0.7 on both ascorbic acids. 相似文献
5.
Michael Ogundare Spyridon Theofilopoulos Andrew Lockhart Leslie J. Hall Ernest Arenas Jan Sj?vall A. Gareth Brenton Yuqin Wang William J. Griffiths 《The Journal of biological chemistry》2010,285(7):4666-4679
In this study we have profiled the free sterol content of cerebrospinal fluid by a combination of charge tagging and liquid chromatography-tandem mass spectrometry. Surprisingly, the most abundant cholesterol metabolites were found to be C27 and C24 intermediates of the bile acid biosynthetic pathways with structures corresponding to 7α-hydroxy-3-oxocholest-4-en-26-oic acid (7.170 ± 2.826 ng/ml, mean ± S.D., six subjects), 3β-hydroxycholest-5-en-26-oic acid (0.416 ± 0.193 ng/ml), 7α,x-dihydroxy-3-oxocholest-4-en-26-oic acid (1.330 ± 0.543 ng/ml), and 7α-hydroxy-3-oxochol-4-en-24-oic acid (0.172 ± 0.085 ng/ml), and the C26 sterol 7α-hydroxy-26-norcholest-4-ene-3,x-dione (0.204 ± 0.083 ng/ml), where x is an oxygen atom either on the CD rings or more likely on the C-17 side chain. The ability of intermediates of the bile acid biosynthetic pathways to activate the liver X receptors (LXRs) and the farnesoid X receptor was also evaluated. The acidic cholesterol metabolites 3β-hydroxycholest-5-en-26-oic acid and 3β,7α-dihydroxycholest-5-en-26-oic acid were found to activate LXR in a luciferase assay, but the major metabolite identified in this study, i.e. 7α-hydroxy-3-oxocholest-4-en-26-oic acid, was not an LXR ligand. 7α-Hydroxy-3-oxocholest-4-en-26-oic acid is formed from 3β,7α-dihydroxycholest-5-en-26-oic acid in a reaction catalyzed by 3β-hydroxy-Δ5-C27-steroid dehydrogenase (HSD3B7), which may thus represent a deactivation pathway of LXR ligands in brain. Significantly, LXR activation has been found to reduce the symptoms of Alzheimer disease (Fan, J., Donkin, J., and Wellington C. (2009) Biofactors 35, 239–248); thus, cholesterol metabolites may play an important role in the etiology of Alzheimer disease. 相似文献
6.
Chih-Tsung Chang Pranav Patel Vivek Gore Wen-Liang Song John A. Lawson William S. Powell Garret A. FitzGerald Joshua Rokach 《Bioorganic & medicinal chemistry letters》2009,19(23):6755-6758
The total and stereospecific synthesis of d4-5-epi-8,12-iso-iPF3α-VI 55 and d4-8,12-iso-iPF3α-VI 64, EPA-derived all-syn-isoprostanes (iPs), has been accomplished. Because of issues related to volatility and yield with some of the primary deuterated synthons an improved synthesis is presented. These two deuterated analogs were used to discover and quantify the presence of the corresponding endogenous isoprostanes in human urine. These assays may serve as a valuable index of oxidative stress in population with omega-3 fatty acid enriched diets containing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and may also be useful as an index of the severity of inflammatory diseases such as atherosclerosis and Alzheimer’s disease. 相似文献
7.
L. Giovagnini L. Ronconi L. Marchiò P. Brun D. Fregona 《Journal of inorganic biochemistry》2009,103(5):774-782
[RuCl3 · nH2O] and Na(trans-[RuCl4(DMSO)2]) were reacted with 1-pyrrolidinedithiocarbamate (PDT), its S-methyl ester (PDTM), and N,N-dimethylcarbamodithioic acid methyl ester (DMDTM) in water or methanol in order to obtain the corresponding Ru(III) derivatives. Once isolated and purified, the complexes were characterized by means of elemental analysis, conductivity measurements, FT-IR and 1H NMR spectroscopy, ion electrospray mass spectrometry (ESI-MS), and thermal analyses. The crystal structure of mer-[Ru(DMDTM)(DMSO)Cl3] has been also determined by X-ray crystallography. In vitro cytotoxic activity of all the synthesized complexes was eventually evaluated on some selected human tumor cell lines. 相似文献
8.
Matthew P. Galloway Bryan L. Roth Carmine J. Coscia 《Archives of biochemistry and biophysics》1981,209(2):620-627
The effects of tetrahydroisoquinolinecarboxylic acids, derived from dopamine and various phenylpyruvates, on the enzyme tyrosine 3-monooxygenase have been investigated. Using a partially purified tyrosine 3-monooxygenase from bovine adrenal medulla, 3′,4′-deoxynorlaudanosolinecarboxylic acid was found to be a mixed inhibitor against the cofactor (Ki = 122 μM), equipotent with norepinephrine. Norlaudanosolinecarboxylic acid inhibited tyrosine 3-monooxygenase competitively with respect to the cofactor (Ki = 126 μM). When tyrosine 3-monooxygenase activity in catecholamine-free striatal homogenates was studied, again 3′,4′-deoxynorlaudanosolinecarboxylic acid (Ki = 40 μM) behaved as a mixed inhibitor whereas norlaudanosolinecarboxylic acid (Ki = 136 μM) was competitive. When the rat striatal tyrosine 3-monooxygenase was subjected to phosphorylating conditions in vitro, decreases in the Ki of norlaudanosolinecarboxylic acid and in that of 3′,4′-deoxynorlaudanosolinecarboxylic acid were observed, whereas the Ki of dopamine was increased. Tyrosine 3-monooxygenase activity in rat striatal synaptosomes was also inhibited by 3′,4′-deoxynorlaudanosolinecarboxylic acid (IC50 = 100 μm) and phosphorylating conditions affected only that inhibition produced by dopamine, but not that by the tetrahydroisoquinolinecarboxylic acids. The results are discussed in relation to the structure of the tetrahydroisoquinolinecarboxylic acids and their possible role in vivo. 相似文献
9.
Deacylation of nitrophenyl acetates containing carboxyl substituents [4-acetoxy-3-nitrobenzoic acid (1), 3-acetoxy-4-nitrobenzoic acid (2), and 2-acetoxy-5-nitrobenzoic acid (3)] was studied in the presence of poly(ethylenimine) derivatives. The polymers examined contained lauryl groups (Lau12PEI) or both lauryl and imidazolyl groups (Lau12Im10PEI). The reaction with active ester proceeds through the attack of primary amino groups of the polymers at the acyl carbons of the substrates. The reaction of Lau12Im10PEI with a hydrophobic ester, p-nitrophenyl caproate (NPC), however, has been reported to involve the attack by the imidazolyl group of the polymer. Thus, the anionic (carboxyl-containing) and the hydrophobic esters bind to different domains on Lau12Im10PEI. Among the anionic substrates, 3 has uniquely large kcat values compared with 1 or 2. This is explained in terms of closer proximity between a nucleophile amino group of the polymer and the scissile bond of the substrate in the polymer-substrate complex. 相似文献
10.
A methyltransferase, which catalyzes the methylation of luteolin (Km, 16 μM) using S-adenosyl-l-methionine as the methyl donor, has been purified about 38-fold from cell suspension cultures of soybean (Glycine max L., var. Mandarin). The following 3,4-dihydroxy phenolic compounds were also methylated: luteolin 7-O-glucoside (Km, 28 μm), quercetin (Km, 35 μm), eriodictyol (Km, 75 μm), 5-hydroxyferulic acid (Km, 227 μm), dihydroquercetin (Km, 435 μm), and caffeic acid (Km, 770 μm). Rutin and quercetin 3-O-glucoside were poor substrates. Methylation proceeded only in the meta position. The enzyme was unable to catalyze the methylation of p-coumaric acid, m-coumaric acid, ferulic acid, isoferulic acid, sinapic acid, apigenin, or naringenin. While the isoflavones biochanin A and daidzein did not serve as substrates, texasin (6,7-dihydroxy-3′-methoxyisoflavone) was methylated (Km, 35 μm). The methylation of caffeic acid and quercetin showed a pH optimum of 8.6–8.9. The enzyme required Mg2+ ions for maximum activity (approximately 1 mm) and could be totally inhibited by EDTA (10 mm). The Km for S-adenosyl-l-methionine was 11 μm. S-Adenosyl-l-homocysteine inhibited the methylation of luteolin by S-adenosyl-l-methionine. 相似文献
11.
Salmonella enterica Serovar Typhimurium and Listeria monocytogenes Acid Tolerance Response Induced by Organic Acids at 20°C: Optimization and Modeling
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E. J. Greenacre T. F. Brocklehurst C. R. Waspe D. R. Wilson P. D. G. Wilson 《Applied microbiology》2003,69(7):3945-3951
An acid tolerance response (ATR) has been demonstrated in Listeria monocytogenes and Salmonella enterica serovar Typhimurium in response to low pH poised (i.e., adapted) with acetic or lactic acids at 20°C and modeled by using dynamic differential equations. The ATR was not immediate or prolonged, and optimization occurred after exposure of L. monocytogenes for 3 h at pH 5.5 poised with acetic acid and for 2 h at pH 5.5 poised with lactic acid and after exposure of S. enterica serovar Typhimurium for 2 h at pH 5.5 poised with acetic acid and for 3 h at pH 5.5 poised with lactic acid. An objective mechanistic analysis of the acid inactivation data yielded estimates of the duration of the shoulder (ts), the log-linear decline (kmax), and the magnitude of a critical component (C). The magnitude of kmax gave the best agreement with estimates of conditions for optimum ATR induction made from the raw data. 相似文献
12.
A 3-O-methyltransferase which catalyzes the methylation of caffeic acid to ferulic acid using S-adenosyl-l-methionine as methyl donor has been isolated and purified about 60-fold from cell suspension cultures of soybean (Glycine max L., var. Mandarin). The enzyme utilized, in addition to caffeic acid (Km = 133 μM), 5-hydroxyferulic acid (Km = 55 μM), 3,4,5-trihydroxy-cinnamic acid (Km = 100 μM), and protocatechualdehyde (Km = 50 μM) as substrates. Methylation proceeded only in the meta position. The enzyme was unable to catalyze the methylation of ferulic acid, of ortho-, meta-, and para-coumaric acids, and of the flavonoid compounds quercetin and luteolin. The methylation of caffeic acid and 5-hydroxyferulic acid showed a pH optimum at 6.5–7.0. No stimulation of the reaction velocity was observed when Mg2+ ions were added. EDTA did not inhibit the reaction. The Km for S-adencsyl-l-methionine was 15 μm. S-Adenosyl-l-homocysteine was a potent competitive inhibitor of S-adenosyl-l-methionine (Ki = 6.9 μM). 相似文献
13.
The action of exogenously applied hormones in the induction of morphogenesis inLimnophila chinensis (Osb.) Merr. tissue culture has been demonstrated. Stem expiants were grown on Murashige and Skoog’s medium containing various levels of kinetin, gibberellic acid and indole-3-acetic acid. Formation of roots, shoots (normal or abnormal), plantlets and friable, hard or nodulated calluses depended largely on the hormone levels used. The formation of normal shoots and roots were stimulated by treatment with kinetin. GA3 treatment stimulated the bud differentiation but inhibited the root initiation. A combination of kinetin and GA3 gave variable results. 相似文献
14.
Guanghua Luo Yuanping Shi Jun Zhang Qinfeng Mu Li Qin Lu Zheng Yuehua Feng Maria Berggren-Söderlund Peter Nilsson-Ehle Xiaoying Zhang Ning Xu 《Biochemical and biophysical research communications》2014
It has been demonstrated that apolipoprotein M (APOM) is a vasculoprotective constituent of high density lipoprotein (HDL), which could be related to the anti-atherosclerotic property of HDL. Investigation of regulation of APOM expression is of important for further exploring its pathophysiological function in vivo. Our previous studies indicated that expression of APOM could be regulated by platelet activating factor (PAF), transforming growth factors (TGF), insulin-like growth factor (IGF), leptin, hyperglycemia and etc., in vivo and/or in vitro. In the present study, we demonstrated that palmitic acid could significantly inhibit APOM gene expression in HepG2 cells. Further study indicated neither PI-3 kinase (PI3K) inhibitor LY294002 nor protein kinase C (PKC) inhibitor GFX could abolish palmitic acid induced down-regulation of APOM expression. In contrast, the peroxisome proliferator-activated receptor beta/delta (PPARβ/δ) antagonist GSK3787 could totally reverse the palmitic acid-induced down-regulation of APOM expression, which clearly demonstrates that down-regulation of APOM expression induced by palmitic acid is mediated via the PPARβ/δ pathway. 相似文献
15.
Leplaeric acid E 5, leplazarin 6a and 21-epileplazarin 6b, three new 3,4-seco-lanostane type triterpenes have been isolated from the stem bark of Leplaea mayombensis (Pellegr.) Staner along with fourteen known compounds from the fruits and roots. Leplaeric acid E, leplazarin and 21-epileplazarin, 15-α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid, mayomlactones A and B, lanosta-7,24-dien-3-one, leplaeric acid A, B and C were screened in vitro for antiplasmodial activity against chloroquine-sensitive (Pf3D7) and chloroquine-resistant (PfINDO) strains of Plasmodium falciparum and for cytotoxicity against CAL-27, CaCo2, Skov-3, and HepG2 cells line. Three compounds including 15-α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid (IC50 5.65–7.09 μM), lanosta-7,24-dien-3-one (IC50 7.18–9.07 μM), and leplaeric acid C (IC50 7.59–8.47 μM) were the most active against both strains of P. falciparum. All the compounds exhibited cytotoxicity against the three-cell lines with IC50 ranging from 12.30 to 181.88 μM. These results confirm the usage of the medicinal plant L. mayombensis for the management of malaria and suggest that further lead optimization studies on potent compounds identified from this study could lead to the identification of potential of lead molecules as scaffold for new antimalarial drug discovery. 相似文献
16.
《Biochimica et Biophysica Acta (BBA)/General Subjects》2001,1526(3):277-285
cis-Urocanic acid (cis-UCA), formed from trans-urocanic acid (trans-UCA) by photoisomerization, has been shown to mimic suppressive effects of UV on the immune system. It is our hypothesis that UCA oxidation products in the skin play a role in the process of immunosuppression. Recently, both UCA isomers were found to be good hydroxyl radical scavengers and in this context we investigated the formation of products resulting from the interaction of hydroxyl radicals with UCA. Hydroxyl radicals were generated by (1) UV/H2O2 (photooxidation), (2) ferrous ions/H2O2 (Fenton oxidation) and (3) cupric ions/ascorbic acid. Oxidation products were identified by spectrometric methods and assessed by reversed-phase HPLC analysis. The photooxidation of UCA was induced by UV-B and UV-C, but not by UV-A radiation. Photooxidation and Fenton oxidation of trans-UCA, as well as of cis-UCA yielded comparable chromatographic patterns of UCA oxidation products. Several of the formed products were identified. The formation of three identified imidazoles was shown in UV-B exposed corneal layer samples, derived from human skin. 相似文献
17.
Prostaglandins (PGs) have been detected in many different plants and certain microorganisms. A few prostaglandin-like compounds have also been shown to occur in plants such as flax,Chromolaena morii, and aquatic sedge; and direct precursors (arachidonic acid, di-homo-γ-linolenic acid and eicosapentaenoic acid) have been detected in a variety of plants and microorganisms, including certain red algae, brown algae, green algae, and saltwater diatoms. Furthermore, arachidonic acid has been found in mosses and a liverwort. It has also been reported that arachidonic acid occurs in certain angiosperms, namely, poplar (Populus balsamifera), wheat germ oil,Aloe vera, andAllium sativum (garlic). In our studies on the possible physiological effects of prostaglandins we found that a PG possibly has an effect on the flowering of the short-day plantPharbitis nil. It has hastened flower formation by 28 days as compared with controls under inductive conditions (short days), and certain inhibitors of PG-biosynthesis inhibited flowering to a greater or lesser extent. In other physiological studies of prostaglandins, it was found that they have an effect on such aspects as GA3-controlled responses in barley endosperm, inhibition of crown gall tumor formation on potato discs, and certain electron-flow reactions in isolated chloroplasts. In corn leaf segments it has an effect on photosynthesis, nucleic acid metabolism, and protein synthesis. The effect on four plant bioassay systems was negligible. It has also been reported that PGs play a role in the regulation of cell membrane permeability. 相似文献
18.
Carbonic anhydrase III from rabbit muscle, a newly discovered major isoenzyme of carbonic anhydrase, has been found to be also a p-nitrophenyl phosphatase, an activity which is not associated with carbonic anhydrases I and II. The p-nitrophenyl phosphatase activity has been shown to chromatograph with the CO2 hydratase activity; both activities are associated with each of its sulfhydryl oxidation subforms; and both activities follow the same pattern of pH stability. This phosphomonoesterase activity of carbonic anhydrase III has an acidic pH optimum (<5.3); its true substrate appears to be the phosphomonoanion with a Km of 2.8 mm. It is competitively inhibited by the typical acid phosphatase inhibitors phosphate (Ki = 1.22 × 10?3M), arsenate (Ki = 1.17 × 10?3M), and molybdate (Ki = 1.34 × 10?7M), with these inhibitors having no effect on the CO2 hydratase or the p-nitrophenyl acetate esterase activities of carbonic anhydrase III. The p-nitrophenyl acetate esterase activity of carbonic anhydrase III, on the other hand, has the sigmoidal pH profile with an inflection at neutral pH, typical of carbonic anhydrases for all of their substrates, and is inhibitable by acetazolamide (a highly specific carbonic anhydrase inhibitor) to the same degree as the CO2 hydratase activity. The acid phosphatase-like activity of carbonic anhydrase III is slightly inhibited by acetazolamide at acidic pH, and inhibited to nearly the same degree at neutral pH. These data are taken to suggest that the phosphatase activity follows a mechanism different from that of the CO2 hydratase and p-nitrophenyl acetate esterase activities and that there is some overlap of the binding sites. 相似文献
19.
Structure and Biosynthesis of Cuticular Lipids: Hydroxylation of Palmitic Acid and Decarboxylation of C(28), C(30), and C(32) Acids in Vicia faba Flowers
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The structure and composition of the cutin monomers from the flower petals of Vicia faba were determined by hydrogenolysis (LiAlH4) or deuterolysis (LiAlD4) followed by thin layer chromatography and combined gas-liquid chromatography and mass spectrometry. The major components were 10, 16-dihydroxyhexadecanoic acid (79.8%), 9, 16-dihydroxyhexadecanoic acid (4.2%), 16-hydroxyhexadecanoic acid (4.2%), 18-hydroxyoctadecanoic acid (1.6%), and hexadecanoic acid (2.4%). These results show that flower petal cutin is very similar to leaf cutin of V. faba. Developing petals readily incorporated exogenous [1-14C]palmitic acid into cutin. Direct conversion of the exogeneous acid into 16-hydroxyhexadecanoic acid, 10, 16-dihydroxy-, and 9, 16-dihydroxyhexadecanoic acid was demonstrated by radio gas-liquid chromatography of their chemical degradation products. About 1% of the exogenous [1-14C]palmitic acid was incorporated into C27, C29, and C31n-alkanes, which were identified by combined gas-liquid chromatography and mass spectrometry as the major components of the hydrocarbons of V. faba flowers. The radioactivity distribution among these three alkanes (C27, 15%; C29, 48%; C31, 38%) was similar to the per cent composition of the alkanes (C27, 12%; C29, 43%; C31, 44%). [1-14C]Stearic acid was also incorporated into C27, C29, and C31n-alkanes in good yield (3%). Trichloroacetate, which has been postulated to be an inhibitor of fatty acid elongation, inhibited the conversion of [1-14C]stearic acid to alkanes, and the inhibition was greatest for the longer alkanes. Developing flower petals also incorporated exogenous C28, C30, and C32 acids into alkanes in 0.5% to 5% yields. [G-3H]n-octacosanoic acid (C28) was incorporated into C27, C29, and C31n-alkanes. [G-3H]n-triacontanoic acid (C30) was incorporated mainly into C29 and C31 alkanes, whereas [9, 10, 11-3H]n-dotriacontanoic acid (C32) was converted mainly to C31 alkane. Trichloroacetate inhibited the conversion of the exogenous acids into alkanes with carbon chains longer than the exogenous acid, and at the same time increased the amount of the direct decarboxylation product formed. These results clearly demonstrate direct decarboxylation as well as elongation and decarboxylation of exogenous fatty acids, and thus constitute the most direct evidence thus far obtained for an elongation-decarboxylation mechanism for the biosynthesis of alkanes. 相似文献
20.
Halina R. Novak Christopher Sayer Jana Panning Jennifer A. Littlechild 《Marine biotechnology (New York, N.Y.)》2013,15(6):695-705
The recombinant l-haloacid dehalogenase from the marine bacterium Psychromonas ingrahamii has been cloned and over-expressed in Escherichia coli. It shows activity towards monobromoacetic (100 %), monochloroacetic acid (62 %), S-chloropropionic acid (42 %), S-bromopropionic acid (31 %), dichloroacetic acid (28 %) and 2-chlorobutyric acid (10 %), respectively. The l-haloacid dehalogenase has highest activity towards substrates with shorter carbon chain lengths (≤C3), without preference towards a chlorine or bromine at the α-carbon position. Despite being isolated from a psychrophilic bacterium, the enzyme has mesophilic properties with an optimal temperature for activity of 45 °C. It retains above 70 % of its activity after being incubated at 65 °C for 90 min before being assayed at 25 °C. The enzyme is relatively stable in organic solvents as demonstrated by activity and thermal shift analysis. The V max and K m were calculated to be 0.6 μM min?1 mg?1 and 1.36 mM with monobromoacetic acid, respectively. This solvent-resistant and stable l-haloacid dehalogenase from P. ingrahamii has potential to be used as a biocatalyst in industrial processes. 相似文献