Isolement, purification et propriétés physico-chimiques des peroxydases de navet

Mazza, Gilbert; Charles, Claudette; Bouchet, Mireille; Ricard, Jacques; Raynaud, Jacques

doi:10.1016/0005-2744(68)90279-9

Cited by 56 publications

(12 citation statements)

References 17 publications

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“…The mobilities of the T12 peptides and their cysteic acid derivatives, and the amino acid compositions of peptides T12(P,) and T12(P2) (where P, and P, indicate peroxidase PI or P,) (Table 6) suggest an amino acid substitution from asparagine in peptide T12(P,) to lysine in T12(P,), at a residue located in the sequence between the two half-cystine residues. The slight differences in molecular weights, carbohydrate and amino acid compositions [2] and absorption spectra [1] of P, and P, all appear to be within experimental error. The differences S ( 1 0 ) 6 ( 8 ) in chromatographic and electrophoretic behaviour.…”

Section: Turnip Pcvwxiduses P and Pmentioning

confidence: 61%

Similarities and Differences of Five Peroxidases from Turnip and Horseradish

Welinder¹,

Mazza²

1975

European Journal of Biochemistry

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Four isoperoxidases of turnip root and isoperoxidase C of horseradish root were digested with trypsin, and their peptide maps, prepared by high-voltage paper electrophoresis, were compared. All five tryptic digests were completely soluble at pH 8. The maps were developed with a variety of general and specific reagents: ninhydrin, histidine, tyrosine, tryptophan and arginine reagents. Cystine peptides and cysteic acid derivatives have also been characterized. All detected half-cystine residues seemed engaged in disulfide bridges. For each individual peroxidase the number of specifically staining peptides agreed very well with the amino acid composition.The two most acidic peroxidases of turnip, PI and P,, only differ significantly in one peptide. The P, gene is tentatively proposed to have developed from the PI gene by a single base mutation, changing an asparagine residue to a lysine residue. A less acidic turnip peroxidase, P3, is distinct, although related to peroxidases P, and P,. Horseradish isoperoxidase C also belongs to this group which appears to be closely related in the amino acid sequences around four disulfide bridges. Peroxidase P, differs from this group, at least around two of its disulfide bridges, and therefore, ma) differ from the other four in parts of its three dimensional structure.Sequences of particular importance to peroxidase function must be present in all peroxidases. From the peptide mapping studies we only find two highly homologous sequences present in all five examined peroxidases. Both contain histidine. This finding corroborates previous suggestions of two histidine sequences near the peroxidase heme prosthetic group.The rules applied in relating peptides of different proteins are outlined, and the sources of errors in mapping of glycoproteins of high carbohydrate content (about 20 %) are discussed in detail.Plant peroxidases are glycoproteins. About 20 ?< of their molecular weight ranging from 40000 to 50000 is carbohydrate. Peroxidases P,, P, and P3 are acid peroxidase isoenzymes (isoelectric points 3.3, 3.5 and 3.7) and peroxidase P, a basic peroxidase isoenzyme (isoelectric point 11.6) of turnip root [l]. They are previously characterized by carbohydrate and amino acid compositions, and molecular weight by Mazza et a/. [2]. Their studies showed that peroxidases P,, P, and P, are similar and that peroxidase P, differs markedly from the three. Horseradish peroxidase isoenzyme C, the major basic peroxidase of horseradish root (isoelectric point 9.0 [3]), is included in the present peptide mapping studies as reference, since its primary structure is nearly completed [3 -71.Ahhreviaizon. Dansyl. 3-dimethyla1ninonaphthalene-5-sulfonyl. Enzymr.c.. Peroxidase (EC 1.1 1.1.7); trypsin (EC 3.4.21.4).Welinder has previously communicated [5,7] that the only two histidine sequences present in horseradish isoperoxidase C, the basic tryptic peptide T8 [3] and the acidic, disulfide-looped tryptic peptide T9 [6], are very similar to the heme-linked histidine sequences of the globin family mono...

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Section: Turnip Pcvwxiduses P and Pmentioning

confidence: 61%

Similarities and Differences of Five Peroxidases from Turnip and Horseradish

Welinder¹,

Mazza²

1975

European Journal of Biochemistry

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“…In this sense, another potentially interesting source of peroxidase is turnip ( Brassica napus L.). Five peroxidase isoenzymes (three anionic and two cationic) have been isolated from turnip roots [7]. Comparison of their amino acid sequences and physicochemical characteristics revealed significant differences both between the isoenzymes themselves and the horseradish enzyme [8,9].…”

Section: Introductionmentioning

confidence: 99%

A peroxidase isoenzyme secreted by turnip (Brassica napus) hairy‐root cultures: inactivation by hydrogen peroxide and application in diagnostic kits

Agostini

Hernández‐Ruíz

Arnao

et al. 2002

Biotech and App Biochem

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We have purified various peroxidase isoenzymes from roots and hairy-root cultures of turnip (Brassica napus) which could potentially be used for commercial applications such as an enzyme immunoassays, diagnostic test kits, wastewater treatment and soil remediation. One of them, a basic peroxidase called HR2, was secreted into the medium of turnip hairy-root cultures. HR2 had a pI of 9.6, a molecular mass of 39.3 kDa and showed great thermostability. The inactivation of HR2 by H2O2 in the absence of reductant substrates was studied. Under these conditions H2O2 acted as a suicide substrate. The kinetic constants calculated have been compared with those of a basic isoperoxidase from horseradish (Armoracia sp.) roots (HRP-C), which is commonly used in commercial kits. The results for HR2 indicated that it was more resistant to inactivation because it presented a lower inactivation efficiency and a higher value for the partition ratio (r=1250) than those described for HRP-C. These results make turnip peroxidase HR2 suitable for use in systems in which high H2O2 concentrations are found. Such an application is demonstrated, namely an enzymic diagnostic kit for determination of uric acid in which HR2 was found to be as efficient as the enzyme originally included in standard kits.

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“…That may be due to difficulties in isolating the peroxidase protein to reasonable purity and amounts. From 240-kg turnip roots, only 53 mg of peroxidase was obtained (Mazza et al 1968), and from petunia, 0.19 mg of peroxidase was obtained from 100 g fresh weight leaves (Hendriks et al 1991). Peanut peroxidase has been purified through extraction from spent medium of cultured peanut cells.…”

Section: Introductionmentioning

confidence: 99%

Isolation of peroxidase from bean and millet compared with that from peanut

Huystee¹,

Malko²,

Wen³

et al. 1994

Can. J. Bot.

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VAN HUYSTEE, R.B., MALKO, M., WEN, K., and GIJZEN, M. 1994. Isolation of peroxidase from bean and millet compared with that from peanut. Can. J. Bot. 72: 1432-1435. Purification of peanut (Arachis hypogaea L.) peroxidase in milligram amounts is feasible through its isolation from spent medium of cultured cells. Using the same and modified approaches for another cultured dicot and a monocot yielded only low amounts of peroxidase for bean (Phaseolus vulgaris) and millet (Setaria italica Beav.). We showed that the procedures used for millet rendered a purer fraction than the commercial crude fraction available for horseradish. The data suggest that cultured peanut cells in suspension culture are an ideal system for isolation of peroxidase in terms of the ease of purification and yield. VAN HUYSTEE, R.B., MALKO, M., WEN, K., et GIJZEN, M. 1994. Isolation of peroxidase from bean and millet comparedwith that from peanut. Can. J. Bot. 72 : 1432-1435. I1 est possible de purifier, en quantitCs de l'ordre du milligramme, la peroxydase de l'arachide (Arachis hypogaea L.) B partir de milieux ayant servi B cultiver ses cellules. Lorsqu'on applique la m&me mCthode ainsi que des variantes B une autre dicotyltdone telle que le haricot (Phaseolus vulgaris) ou une monocotylCdone telle que le millet (Setaria italica Beav.), on obtient que de petites quantitts de peroxydase. Les auteurs constatent que ces mCthodes, appliquCes au millet, permettent d'obtenir une fraction plus pure que la fraction brute produite commercialement B partir du raifort. Les donnCes suggkrent que les cellules d'arachide cultivtes en suspension constituent un systkme idCal pour isoler la peroxydase, considerant les rendements obtenus et la facilitC de la purifier.

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Isolement, purification et propriétés physico-chimiques des peroxydases de navet

Cited by 56 publications

References 17 publications

Similarities and Differences of Five Peroxidases from Turnip and Horseradish

Similarities and Differences of Five Peroxidases from Turnip and Horseradish

A peroxidase isoenzyme secreted by turnip (Brassica napus) hairy‐root cultures: inactivation by hydrogen peroxide and application in diagnostic kits

Isolation of peroxidase from bean and millet compared with that from peanut

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