Biomedical subjects

M G Boersma

Publications and source records attributed to M G Boersma.

At least 19 recordsLinked to original sources

PubMed2007

Digestibility of energy and phosphorus in ten samples of distillers dried grains with solubles fed to growing pigs.

An experiment was conducted to measure DE and ME and the apparent total tract digestibility (ATTD) of energy, N, and P in distillers dried grains with solubles (DDGS) fed to growing pigs. Ten sources of DDGS were obtained from ethanol plants in South Dakota and Minnesota, and 11 diets were formulated. One diet was based on corn (96.8%), limestone, salt, vitamins, and microminerals. Ten additional diets were formulated by mixing the corn diet and each of the 10 sources of DDGS in a 1:1 ratio. Eleven growing pigs (initial BW of 29.3 +/- 0.42 kg) were allotted to an 11 x 11 Latin square design, with 11 periods and 11 pigs. Each of the 11 diets was fed to each pig during 1 period. Pigs were placed in metabolism cages that allowed for the total, but separate, collection of feces and urine. Samples were analyzed for GE, N, and P and energy and N balances, and the ATTD of GE, N, and P were calculated for each diet. By subtracting the contribution from the corn diet to the DDGS-containing diets, the energy and N balances and the ATTD for GE, N, and P for each source of DDGS were calculated. Results of the experiment showed that the DE and ME differed (P < 0.001) among the 10 sources of DDGS (3,947 to 4,593 kcal of DE/kg of DM and 3,674 to 4,336 kcal of ME/kg of DM). The average DE and ME in DDGS were 4,140 and 3,897 kcal/kg of DM, respectively. These values were not different from the DE and ME in corn (4,088 and 3,989 kcal/kg of DM, respectively). Based on the analyzed GE and nutrient composition of DDGS and the calculated values for DE and ME, prediction equations for DE and ME were developed. These equations showed that DE and ME in DDGS may be predicted from the concentration of ash, ether extract, ADF, and GE. The retention of N from DDGS was greater (P < 0.001) than from corn, but when calculated on a percentage basis, the N retention did not differ between DDGS and corn. The ATTD of P in DDGS was 59.1% on average for the 10 samples. This value was greater (P < 0.001) than the ATTD of P in corn (19.3%). It is concluded that the DE and ME in DDGS is not different from the DE and ME in corn. However, if DDGS is included in diets fed to growing swine, a greater portion of the organic P will be digested and absorbed, thus reducing the need for adding inorganic P to the diets.

Animal Feed↗

PubMed2006

Amino acid and energy digestibility in ten samples of distillers dried grain with solubles fed to growing pigs.

The objective of this experiment was to measure the digestibilities of energy, CP, and AA in 10 samples of corn distillers dried grain with solubles (DDGS) and in corn fed to growing pigs. Twelve growing barrows (initial BW: 34.0 +/- 1.41 kg) were allotted to an 8 x 12 Youden square design with 8 periods and 12 animals. Ten of 12 diets were based on the 10 DDGS samples (66.7%), 1 diet was based on corn (97%), and the last diet was a N-free diet based on cornstarch and sucrose. Chromic oxide (0.3%) was included in all diets as an inert marker. Pigs were provided their respective diets at a level of 3 times their estimated energy requirement for maintenance. The apparent (AID) and standardized (SID) ileal digestibilities for CP and AA were measured in the 10 samples of DDGS and in corn using the direct procedure, but the apparent total tract digestibilities for DM and GE were estimated using the difference procedure. The concentration of DE in each sample of DDGS and in corn was also calculated. The results of the experiment indicated variation among the different sources of DDGS in AID and SID for Lys, which ranged from 35.0 to 55.9% and 43.9 to 63.0%, respectively. For Met, the SID varied between 73.9 and 84.7%. However, the variability among samples in the SID for CP, and for the indispensable AA other than Lys and Met, was relatively low and ranged between 6 and 8 percentage units (i.e., from 64.0 to 70.6%, 74.1 to 80.1%, and 67.4 to 75.3% for Thr, Trp, and Ile, respectively). The SID for Trp in corn (72.8%) was lower (P < 0.05) than in DDGS, but for the remaining indispensable AA, except Arg, the SID for corn were greater (P < 0.01) than for DDGS. The DE concentration in the 10 samples of DDGS varied (P < 0.001) from 3,382 to 3,811 kcal of DE per kg of DM. For corn, the DE was 3,845 kcal per kg of DM. It is concluded that the AID and SID for Lys vary among samples of DDGS, but for most other AA the AID and SID are relatively similar and vary only 6 to 8 percentage units among different samples. Future work should focus on identifying the reasons for the variation in the digestibility of Lys to avoid processing procedures that are detrimental to Lys digestibility.

Amino Acids↗

PubMed2001

Structure activity relationships for the chemical behaviour and toxicity of electrophilic quinones/quinone methides.

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Benzoquinones↗

PubMed2001

Structure-activity study on the quinone/quinone methide chemistry of flavonoids.

A structure-activity study on the quinone/quinone methide chemistry of a series of 3',4'-dihydroxyflavonoids was performed. Using the glutathione trapping method followed by HPLC, (1)H NMR, MALDI-TOF, and LC/MS analysis to identify the glutathionyl adducts, the chemical behavior of the quinones/quinone methides of the different flavonoids could be deduced. The nature and type of mono- and diglutathionyl adducts formed from quercetin, taxifolin, luteolin, fisetin, and 3,3',4'-trihydroxyflavone show how several structural elements influence the quinone/quinone methide chemistry of flavonoids. In line with previous findings, glutathionyl adduct formation for quercetin occurs at positions C6 and C8 of the A ring, due to the involvement of quinone methide-type intermediates. Elimination of the possibilities for efficient quinone methide formation by (i) the absence of the C3-OH group (luteolin), (ii) the absence of the C2=C3 double bond (taxifolin), or (iii) the absence of the C5-OH group (3,3',4'-trihydroxyflavone) results in glutathionyl adduct formation at the B ring due to involvement of the o-quinone isomer of the oxidized flavonoid. The extent of di- versus monoglutathionyl adduct formation was shown to depend on the ease of oxidation of the monoadduct as compared to the parent flavonoid. Finally, unexpected results obtained with fisetin provide new insight into the quinone/quinone methide chemistry of flavonoids. The regioselectivity and nature of the quinone adducts that formed appear to be dependent on pH. At pH values above the pK(a) for quinone protonation, glutathionyl adduct formation proceeds at the A or B ring following expected quinone/quinone methide isomerization patterns. However, decreasing the pH below this pK(a) results in a competing pathway in which glutathionyl adduct formation occurs in the C ring of the flavonoid, which is preceded by protonation of the quinone and accompanied by H(2)O adduct formation, also in the C ring of the flavonoid. All together, the data presented in this study confirm that quinone/quinone methide chemistry can be far from straightforward, but the study provides significant new data revealing an important pH dependence for the chemical behavior of this important class of electrophiles.

Benzoquinones↗

PubMed2001

19F NMR metabolomics for the elucidation of microbial degradation pathways of fluorophenols.

Of all NMR-observable isotopes (19)F is the one most convenient for studies on the biodegradation of environmental pollutants and especially for fast initial metabolic screening of newly isolated organisms. In the past decade we have identified the (19)F NMR characteristics of many fluorinated intermediates in the microbial degradation of fluoroaromatics including especially fluorophenols. In the present paper we give an overview of results obtained for the initial steps in the aerobic microbial degradation of fluorophenols, i.e. the aromatic hydroxylation to di-, tri- or even tetrahydroxybenzenes ultimately suitable as substrates for the second step, ring cleavage by dioxygenases. In addition we present new results from studies on the identification of metabolites resulting from reaction steps following aromatic ring cleavage, i.e. resulting from the conversion of fluoromuconates by chloromuconate cycloisomerase. Together the presented data illustrate the potential of the (19)F NMR technique for (1) fast initial screening of biodegradative pathways, i.e. for studies on metabolomics in newly isolated microorganisms, and (2) identification of relatively unstable pathway intermediates like fluoromuconolactones and fluoromaleylacetates.

Journal Article↗

PubMed2001

Transformation of the insecticide teflubenzuron by microorganisms.

Transformation of teflubenzuron, the active component in the insecticide commercialized as Nomolt, by soil microorganisms was studied. It was shown that microorganisms, belonging to Bacillus, Alcaligenes, Pseudomonas and Acinetobacter genera are capable to perform the hydrolytic cleavage of the phenylurea bridge of teflubenzuron in different positions, especially active was Bacillus brevis 625. The structure of the intermediates formed was established using TLC, HPLC, mass-spectrometry and 19F NMR techniques. It was shown that for a dose range of 53-132 microM and upon 12 days of fermentation about 30% of the teflubenzuron was modified. About 10-15% was transformed into 2,6-difluorobenzamide, 3-5% into 2,6-difluorobenzoic acid, 10-12% into 2,4-difluoro-3,5-dichloro-aniline. The late compound gave rise to formation of a condensed compound, identified as 1,2-bis(2,4-difluoro-3,5-dichlorophenyl)urea with molecular mass of 420. The results obtained indicate degradation of teflubenzuron by soil microorganisms to be a process to be mediated by microbial consortia, and starting with hydrolysis of the phenylurea bridge by several bacterial species. Subsequent further degradation of the aromatic degradation products has to be mediated by other strains known to be capable of degradation of halogenated aromatics.

Acinetobacter calcoaceticus↗

PubMed2000

Peroxidase-catalyzed formation of quercetin quinone methide-glutathione adducts.

The oxidation of quercetin by horseradish peroxidase/H(2)O(2) was studied in the absence but especially also in the presence of glutathione (GSH). HPLC analysis of the reaction products formed in the absence of GSH revealed formation of at least 20 different products, a result in line with other studies reporting the peroxidase-mediated oxidation of flavonoids. In the presence of GSH, however, these products were no longer observed and formation of two major new products was detected. (1)H NMR identified these two products as 6-glutathionylquercetin and 8-glutathionylquercetin, representing glutathione adducts originating from glutathione conjugation at the A ring instead of at the B ring of quercetin. Glutathione addition at positions 6 and 8 of the A ring can best be explained by taking into consideration a further oxidation of the quercetin semiquinone, initially formed by the HRP-mediated one-electron oxidation, to give the o-quinone, followed by the isomerization of the o-quinone to its p-quinone methide isomer. All together, the results of the present study provide evidence for a reaction chemistry of quercetin semiquinones with horseradish peroxidase/H(2)O(2) and GSH ultimately leading to adduct formation instead of to preferential GSH-mediated chemical reduction to regenerate the parent flavonoid.

Chromatography, High Pressure Liquid↗

PubMed2000

Characterization of different commercial soybean peroxidase preparations and use of the enzyme for N-demethylation of methyl N-methylanthranilate To produce the food flavor methylanthranilate.

The potential of different peroxidase preparations for the N-demethylation of methyl N-methylanthranilate to produce the food flavor methylanthranilate (MA) was investigated. All tested peroxidase preparations were able to catalyze the N-dealkylation. The tested soybean preparations vary widely with respect to their heme content. Furthermore, the operational stability of purified soybean peroxidase (SP) is at least 25-fold lower than that of horseradish peroxidase and only 5-fold higher than that of microperoxidase 8. Thus, the presence of a large protein chain around a porphyrin cofactor in a peroxidase is, by itself, insufficient to explain the observed differences in operational stability. Despite its relatively low operational stability, SP proved to be the most efficient biocatalyst for the production of MA with high yield and purity, especially observed at the high temperature and low pH values at which SP appeared to be optimally active.

Flavoring Agents↗

PubMed2000

Regioselectivity and reversibility of the glutathione conjugation of quercetin quinone methide.

The chemical reactivity, isomerization, and glutathione conjugation of quercetin o-quinone were investigated. Tyrosinase was used to generate the unstable quercetin o-quinone derivative which could be observed upon its subsequent scavenging by glutathione. Identification of the products revealed formation of 6-glutathionyl-quercetin and 8-glutathionyl-quercetin adducts. Thus, in particular, glutathione adducts in the A ring of quercetin were formed, a result which was not expected a priori. Quantum mechanical calculations support the possibility that the formation of these glutathione adducts can be explained by an isomerization of quercetin o-quinone to p-quinone methides. Surprisingly, additional experiments of this study reveal the adduct formation to be reversible, leading to interconversion between the two quercetin glutathione adducts and possibilities for release and further electrophilic reactions of the quercetin quinone methide at cellular sites different from those of its generation.

Chromatography, High Pressure Liquid↗

PubMed2000

Heme-(hydro)peroxide mediated O- and N-dealkylation. A study with microperoxidase.

The mechanism of microperoxidase-8 (MP-8) mediated O- and N-dealkylation was investigated. In the absence of ascorbate (peroxidase mode), many unidentified polymeric products are formed and the extent of substrate degradation correlates (r = 0.94) with the calculated substrate ionization potential, reflecting the formation of radical intermediates. In the presence of ascorbate (P450 mode) formation of polymeric products is largely prevented but, surprisingly, dealkylation is not affected. In addition, aromatic hydroxylation and oxidative dehalogenation is observed. The results exclude a radical mechanism and indicate the involvement of a (hydro)peroxo-iron heme intermediate in P450-type of heteroatom dealkylation.

Alkylation↗

PubMed2000

Identification of fluoropyrogallols as new intermediates in biotransformation of monofluorophenols in Rhodococcus opacus 1cp.

The transformation of monofluorophenols by whole cells of Rhodococcus opacus 1cp was investigated, with special emphasis on the nature of hydroxylated intermediates formed. Thin-layer chromatography, mass spectrum analysis, and (19)F nuclear magnetic resonance demonstrated the formation of fluorocatechol and trihydroxyfluorobenzene derivatives from each of three monofluorophenols. The (19)F chemical shifts and proton-coupled splitting patterns of the fluorine resonances of the trihydroxyfluorobenzene products established that the trihydroxylated aromatic metabolites contained hydroxyl substituents on three adjacent carbon atoms. Thus, formation of 1,2, 3-trihydroxy-4-fluorobenzene (4-fluoropyrogallol) from 2-fluorophenol and formation of 1,2,3-trihydroxy-5-fluorobenzene (5-fluoropyrogallol) from 3-fluorophenol and 4-fluorophenol were observed. These results indicate the involvement of fluoropyrogallols as previously unidentified metabolites in the biotransformation of monofluorophenols in R. opacus 1cp.

Biotransformation↗

PubMed1999

Preferential oxidative dehalogenation upon conversion of 2-halophenols by Rhodococcus opacus 1G.

The regiospecificity of hydroxylation of C2-halogenated phenols by Rhodococcus opacus 1G was investigated. Oxidative defluorination at the C2 position ortho with respect to the hydroxyl moiety was preferred over hydroxylation at the non-fluorinated C6 position for all 2-fluorophenol compounds studied. Initial hydroxylation of 2,3, 5-trichlorophenol resulted in the exclusive formation of 3, 5-dichlorocatechol. These results indicate that, in contrast to all other phenol ortho-hydroxylases studied so far, phenol hydroxylase from R. opacus 1G is capable of catalyzing preferential oxidative defluorination but also oxidative dechlorination.

Biodegradation, Environmental↗

PubMed1999

Differential substrate behaviour of phenol and aniline derivatives during conversion by horseradish peroxidase.

For the first time saturating overall k(cat) values for horseradish peroxidase (HRP) catalysed conversion of phenols and anilines are described. These k(cat) values correlate quantitatively with calculated ionisation potentials of the substrates. The correlations for the phenols are shifted to higher k(cat) values at similar ionisation potentials as compared to those for anilines. (1)H-NMR T(1) relaxation studies, using 3-methylphenol and 3-methylaniline as the model substrates, revealed smaller average distances of the phenol than of the aniline protons to the paramagnetic Fe(3+) centre in HRP. This observation, together with a possibly higher extent of deprotonation of the phenols than of the anilines upon binding to the active site of HRP, may contribute to the relatively higher HRP catalysed conversion rates of phenols than of anilines.

Aniline Compounds↗

PubMed1999

The effect of iron to manganese substitution on microperoxidase 8 catalysed peroxidase and cytochrome P450 type of catalysis.

This study describes the catalytic properties of manganese microperoxidase 8 [Mn(III)MP8] compared to iron microperoxidase 8 [Fe(III)MP8]. The mini-enzymes were tested for pH-dependent activity and operational stability in peroxidase-type conversions, using 2-methoxyphenol and 3,3'-dimethoxybenzidine, and in a cytochrome P450-like oxygen transfer reaction converting aniline to para-aminophenol. For the peroxidase type of conversions the Fe to Mn replacement resulted in a less than 10-fold decrease in the activity at optimal pH, whereas the aniline para-hydroxylation is reduced at least 30-fold. In addition it was observed that the peroxidase type of conversions are all fully blocked by ascorbate and that aniline para-hydroxylation by Fe(III)MP8 is increased by ascorbate whereas aniline para-hydroxylation by Mn(III)MP8 is inhibited by ascorbate. Altogether these results indicate that different types of reactive metal oxygen intermediates are involved in the various conversions. Compound I/II, scavenged by ascorbate, may be the reactive species responsible for the peroxidase reactions, the polymerization of aniline and (part of) the oxygen transfer to aniline in the absence of ascorbate. The para-hydroxylation of aniline by Fe(III)MP8, in the presence of ascorbate, must be mediated by another reactive iron-oxo species which could be the electrophilic metal(III) hydroperoxide anion of microperoxidase 8 [M(III)OOH MP8]. The lower oxidative potential of Mn, compared to Fe, may affect the reactivity of both compound I/II and the metal(III) hydroperoxide anion intermediate, explaining the differential effect of the Fe to Mn substitution on the pH-dependent behavior, the rate of catalysis and the operational stability of MP8.

Catalysis↗

PubMed1999

TEAC antioxidant activity of 4-hydroxybenzoates.

The influence of pH, intrinsic electron donating capacity, and intrinsic hydrogen atom donating capacity on the antioxidant potential of series of hydroxy and fluorine substituted 4-hydroxybenzoates was investigated experimentally and also on the basis of computer calculations. The pH-dependent behavior of the compounds in the TEAC assay revealed different antioxidant behavior of the nondissociated monoanionic form and the deprotonated dianionic form of the 4-hydroxybenzoates. Upon deprotonation the radical scavenging ability of the 4-hydroxybenzoates increases significantly. For mechanistic comparison a series of fluorobenzoates was synthesized and included in the studies. The fluorine substituents were shown to affect the proton and electron donating abilities of 4-hydroxybenzoate in the same way as hydroxyl substituents. In contrast, the fluorine substituents influenced the TEAC value and the hydrogen atom donating capacity of 4-hydroxybenzoate in a way different from the hydroxyl moieties. Comparison of these experimental data to computer-calculated characteristics indicates that the antioxidant behavior of the monoanionic forms of the 4-hydroxybenzoates is not determined by the tendency of the molecule to donate an electron, but by its ability to donate a hydrogen atom. Altogether, the results explain qualitatively and quantitatively how the number and position of OH moieties affect the antioxidant behavior of 4-hydroxybenzoates.

Antioxidants↗

PubMed1999

Antioxidant activities of carotenoids: quantitative relationships between theoretical calculations and experimental literature data.

Quantitative structure activity relationships (QSARs) are described for the antioxidant activity of series of all-trans carotenoids. The antioxidant activity of the carotenoids is characterised by literature data for (i) their relative ability to scavenge the ABTS*+ radical cation, reflected by the so-called trolox equivalent antioxidant capacity (TEAC) value, (ii) their relative rate of oxidation by a range of free radicals, or (iii) their capacity to inhibit lipid peroxidation in multilamellar liposomes, leading to a decrease in formation of thiobarbituric acid reactive substances (TBARS). All these antioxidant values for radical scavenging action correlate quantitatively with computer-calculated ionisation potentials of the carotenoids. These correlations are observed both when the ionisation potential is calculated as the negative of the energy of the highest occupied molecular orbital (-E(HOMO)) of the molecule, or as the relative change in heat of formation (deltadeltaHF) upon the one-electron oxidation of the carotenoids. The calculations provide a theoretical assay able to characterise the intrinsic electron donating capacity of an antioxidant, in hydrophilic, hydrophobic or artificial membrane environment.

Antioxidants↗

PubMed1998

Quantitative structure/activity relationship for the rate of conversion of C4-substituted catechols by catechol-1,2-dioxygenase from Pseudomonas putida (arvilla) C1.

The influence of various C4/C5 substituents in catechol (1,2-dihydroxybenzene) derivatives on the overall rate of conversion by catechol-1,2-dioxygenase from Pseudomonas putida (arvilla) C1 was investigated. Using catechol, 4-methylcatechol, 4-fluorocatechol, 4-chlorocatechol, 4-bromocatechol, 4,5-difluorocatechol and 4-chloro-5-fluorocatechol, it could be demonstrated that substituents at the C4 and/or C5 position decrease the rate of conversion, from 62% (4-methylcatechol) down to 0.7% (4-chloro-5-fluorocatechol) of the activity with non-substituted catechol. The inhibition was reversible upon addition of excess catechol for all substrates tested. This indicates that the lower activities are neither due to irreversible inactivation of the enzyme nor to product inhibition. Based on the reaction mechanism proposed in the literature [Que, L. & Ho, R. Y. N. (1996) Chem. Rev. 96, 2606-2624], the nucleophilic reactivity of the catecholate was expected to be an essential characteristic for its conversion by catechol-1,2-dioxygenase. Therefore, the rates of conversion were compared with calculated energies of the highest occupied molecular orbital (E(HOMO)) of the substrates. A clear quantitative relationship (R>0.97) between the ln kcat and the calculated electronic parameter E(HOMO) was obtained. This indicates that the rate-limiting step of the reaction cycle is dependent on the nucleophilic reactivity of the substrate and not sterically hindered by the relatively large bromine or methyl substituents used in the present study. Possible steps in the reaction mechanism determining the overall rate at 20 degrees C are discussed.

Catechol 1,2-Dioxygenase↗

PubMed1998

Oxygen exchange with water in heme-oxo intermediates during H2O2-driven oxygen incorporation in aromatic hydrocarbons catalyzed by microperoxidase-8.

The present paper describes the oxygen incorporation into naphthalene and anthracene by H2O2-driven microperoxidase-8, forming alpha-naphthol and anthraquinone, respectively. Microperoxidase-8 is a minienzyme containing a histidinyl-coordinated Fe3+-protoporphyrin IX cofactor covalently attached to an eight-amino-acid peptide. Additional experiments were performed to investigate whether the reaction mechanism involved is like that of peroxidase and/or cytochrome P-450. A reaction pathway like that of cytochrome P-450 implies oxygen transfer to the substrate from the as yet uncharacterized iron-oxo species formed in the reaction of the heine cofactor with H2O2. In contrast, a peroxidase-type reaction chemistry involves reaction pathways proceeding by initial one-electron oxidation of, or H-abstraction from, the substrate, followed by incorporation of oxygen from sources other than the iron-oxo species, i.e. from other than H2O2. The results of the present study exclude Fenton-type chemistry and prove that the minicatalyst is able to catalyze the oxygen incorporation by both peroxidase and cytochrome P-450 types of reaction pathways, while exchange occurs between the high-valency iron-oxo species and H2O. The mechanistic implications of this exchange for cytochrome P-450 are discussed.

Animals↗