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1.
Sabine Lhernould Yannis Karamanos Sylvain Bourgerie Gerard Strecker Raymond Julien Henri Morvan 《Glycoconjugate journal》1992,9(4):191-197
We have previously isolated mannoside and xylomannoside oligosaccharides with one or two terminal reducingN-acetylglucosamine residues from the extracellular medium of white campion (Silene alba) suspension culture. We have now demonstrated the presence of peptide-N
4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity in cell extracts as well in the culture medium that could explain the production of those compounds. An additional xylomannoside, (GlcNAc)Man3(Xyl)GlcNAc(Fuc)GlcNAc, was characterized, and1H- and13C-NMR assignments for the oligosaccharide Man3(Xyl)GlcNAc(Fuc)GlcNAc were obtained using homonuclear and heteronuclear spectroscopy (COSY).Abbreviations Endo
endo--N-acetylglucosaminidase
- Fuc
fucose
- GlcNAc
N-acetylglucosamine
- Man
mannose
- NMR
nuclear magnetic resonance
- PNGase
peptide-N
4-(N-acetylglucosaminyl)asparagine amidase
- Xyl
xylose 相似文献
2.
Sylvain Bourgerie Yannis Karamanos Sylvie Berger Raymond Julien 《Glycoconjugate journal》1992,9(4):162-167
Peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase F (PNGase F) and endo--N-acetyl glucosaminidase F (Endo F) activities were monitored during cultivation ofFlavobacterium meningosepticum using a new fluorescence-HPLC procedure based on a commercially available substrate. The PNGase F activity reached a maximum level at the end of the log phase and remained constant during the stationary phase, while Endo F continuously increased until late stationary phase. PNGase F obtained at the end of the log phase was less contaminated by other proteins compared with late stationary phase.Abbreviations Con A
concanavalin A
- Endo F
endo--N-acetyl glucosaminidase F (EC 3.2.1.96)
- GlcNAc
N-acetylglucosamine
- PNGase F
peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase F (EC 3.5.1.52). 相似文献
3.
Günter Pfeiffer Dietmar Linder Karlhermann Strube Rudolf Geyer 《Glycoconjugate journal》1993,10(3):240-246
In a previous study, we determined the structures of the glycans present in ancrod, a thrombin-like serine protease from the venom of the Malayan pit viperAgkistrodon rhodostoma (Pfeifferet al. (1992)Eur J Biochem
205:961–78). In order to allocate the various carbohydrate chains to distinctN-glycosylation sites of the molecule, we have now isolated individual glycopeptides. Peptide moieties were identified after deglycosylation with peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase F by amino acid analysis and sequencing. Liberated oligosaccharides were assigned to the previously deduced carbohydrate structures by high performance liquid chromatography. Although only quantitative differences were observed, the results indicate that each glycosylation site of ancrod carries its characteristic oligosaccharide pattern. Furthermore, all potential sites were shown to be substituted by carbohydrates.Abbreviations HPAE-HPLC
high pH anion exchange HPLC
- RP-HPLC
reversed phase HPLC
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase F
- PAD
pulsed amperometric detection. 相似文献
4.
Jan B L Damm Hans Voshol Karl Hård Johannis P Kamerling Gijs W K Van Dedem Johannes F G Vliegenthart 《Glycoconjugate journal》1988,5(3):221-233
TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz1H-NMR spectroscopy to beAbbreviations hCG
human chorionic gonadotropin
- hCG-
-subunit
- hCG-
-subunit
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52)
- endo-F
endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96)
- SDS
sodium dodecyl sulphate
- PAGE
polyacrylamide gel electrophoresis
- CBB
coomassie brilliant blue R 250
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose 相似文献
5.
《Bioscience, biotechnology, and biochemistry》2013,77(2):412-418
We report here the isolation and characterization of a peptide-N 4-(acetyl-β-glucosaminyl) asparagine amidase (peptide: N-glycanase) from soybean (Glycine max) seeds. The enzyme was purified to homogeneity with 6.5% yield from defatted soybean meal extract by ion-exchange chromatography, gel filtration, hydroxyapatite chromatography, and hydrophobic chromatography. The purified enzyme, designated PNGase-GM, had the apparent molecular mass of 93 kDa by SDS-PAGE and 90 kDa by gel filtration, indicating this PNGase is a monomeric protein. The enzyme showed maximal activity at pH 4.5-5.0. PNGase-GM was capable of hydrolyzing the β-aspartylglycosylamine linkage (GlcNAcβ1→Asn) of various glycopeptide substrates bearing high-mannose type, hybrid type, and xylose/fucose-containing plant complex type N-glycan units, while this amidase was far less active on the glycopeptides bearing sialylated animal complex-type glycans. 相似文献
6.
Recently, we have reported purification and characterization of a de-N-glycosylating enzyme, peptide:N-glycanase (PNGase) found in C3H mouse fibroblast L-929 cells, and designated L-929 PNGase [Suzuki T, Seko A, Kitajima K, Inoue Y, Inoue S (1994)J Biol Chem
269, 17611–18]. The unique properties of L-929 PNGase are that the enzyme had a high affinity to the substrate glycopeptide (e.g.K
m=114 µm for fetuin derived glycopentapeptide) and that the PNGase-catalysed reaction is strongly inhibited by the released free oligosaccharides but not by the free peptides formed, suggesting that L-929 PNGase is able to bind to a certain type of carbohydrate chain. In this study, we report the new findings of the mannan-binding property of L-929 PNGase; the de-N-glycosylating enzyme activity of L-929 PNGase was inhibited by yeast mannan and triomannose, Man1 3(Man1 6)Man, but not by mannose and -methyl-d-mannoside. Furthermore, L-929 PNGase was revealed to bind to the glycan moiety of yeast mannan by using mannan-conjugated Sepharose 4B gel as a ligand, suggesting that L-929 PNGase could serve not only as an enzyme but also as a carbohydrate recognition proteinin vivo. Such dual properties found for animal-derived L-929 PNGase are unique and are not shared with other previously characterized plant- and bacterial-origin PNGases — PNGase A and PNGase F, respectively.Abbreviations GLC
gas liquid chromatography
- GlcNAc-Asn
2-acetamido-1--(l-aspartamido)-1,2-dideoxy-d-glucose
- BSA
bovine serum albumin
- DTT
dithiothreitol
- EDTA
ethylenediaminetetraacetic acid
- Gal
d-galactose
- GlcNAc
N-acetyl-d-glucosamine
- Man
d-mannose; triomannose, Man1 3(Man1 6)Man;
- MES
2-(N-morphorino)ethanesulfonic acid
- NeuAc
N-acetyl-neuraminic acid
- PNGase
peptide:N
4-(N-accetyl-glucosaminyl)asparagine amidase (peptide:N-glycanase,EC 3.5.1.52)
- PNP
p-nitrophenyl 相似文献
7.
Peptide-N4-(N-acetyl--glucosaminyl)asparagine amidase F (PNGase F) fromFlavobacterium meningosepticum is a highly useful enzyme for the structural analysis of N(asparagine)-linked carbohydrate chains derived from glycoproteins. The enzyme was enriched using a published procedure [Tarentino AL, Gomez CM, Plummer TH, Jr (1984) Biochemistry 1985:4665–71; Tarentino AL, Plummer TH, Jr (1987) Methods Enzymol 138:770–78] and further purified by hydrophobic interaction HPLC on a weak hydrophobic TSK-Ether column from which it was eluted by a decreasing gradient of 1.7 M ammonium sulphate in 100 mM sodium phosphate, pH 7.0, containing 5 mM EDTA.To determine the optimal conditions for a complete deglycosylation of glycoproteins by PNGase F, experiments were performed with human
1-acid glycoprotein, because the five complex type carbohydrate chains are quite resistant to enzymic hydrolysis. The influence of different detergents on the enzyme reaction was studied. Complete deglycosylation of human
1-acid glycoprotein was achieved by the use of 60 mU/ml PNGase F in 0.25 M sodium phosphate buffer, pH 8.6, containing 0.2% (w/v) SDS, 20 mM mercaptoethanol and 0.5% Mega-10. 相似文献
8.
Gabriele D'andrea Jan B Bouwstra Johannis P Kamerling Johannes F G Vliegenthart 《Glycoconjugate journal》1988,5(2):151-157
Ascorbic acid oxidase (E.C.1.10.3.3) from the green zucchini squash (Cucurbita pepo medullosa) is a copper-containing glycoprotein which catalyzes the reaction:l-ascorbic acid +1/2 O2l-dehydroascorbic acid + H2O. The carbohydrate content of the purified plant glycoprotein amounted to 3% (w/w), and monosaccharide analysis revealed the carbohydrate moiety to be of theN-glycosidic type. The carbohydrate chains were released from the apoenzyme by digestion with PNGase-F immobilized on Sepharose 4B. After fractionation on Bio-Gel P-2 and purification on Mono-Q, the neutral oligosaccharide was investigated by 500-MHz1H-NMR spectroscopy. The primary structure of theN-linked carbohydrate chain was established to be:
Abbreviations AAO
ascorbic acid oxidase
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F
- GalNAc
N-acetylgalactosamine
- GlcNAc
N-acetylglucosamine
- Man
mannose
- Xyl
xylose
- GLC
gas-liquid chromatography
- FPLC
fast protein liquid chromatography
- NMR
nuclear magnetic resonance
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
9.
Céline Faugeron Jean-Claude Mollet Yannis Karamanos Henri Morvan 《Acta Physiologiae Plantarum》2006,28(6):557-565
Activities of two de-N-glycosylation enzymes, PNGase (peptide N
4(N-acetyl-glucosaminyl)asparagine amidase) and ENGase (endo N-acetyl-β-D-glucosaminidase), involved in the release of N-glycans from N-glycoproteins, were monitored in several organs of tomato plants (Lycopersicon esculentum, Mill., cv. Dombito) with a fluorescence-HPLC procedure using a resofurin-labelled N-glycopeptide substrate. PNGase and ENGase
activities were detected in every organ assayed but with quantitative differences. The highest activities were found in the
youngest parts of the plant, i.e. apical buds, flowers and leaf blades. PNGase activities were consistently higher than ENGase
activities (three-fold in average). Both de-N-glycosylation activities were associated with high levels of proteins and protease activities. During fruit growth and ripening,
these three parameters decreased notably. The ubiquitous detection of these enzyme activities in the different organs is probably
associated with the previously characterized unconjugated N-glycans in tomato. The possible role of PNGase and ENGase degradation
products (i.e. unconjugated N-glycans) are discussed in relation with their biological functions in plant development. 相似文献
10.
Gabriele D''Andrea Anna M. D''Alessandro M. Luisa Salucci Arduino Oratore 《Journal of Protein Chemistry》1994,13(1):31-36
Human seminal transferrin (HSmT) is an iron-containing glycoprotein whose structural properties have not been adequately investigated. The carbohydrate content of the purified glycoprotein amount to 6.1%, and monosaccharide analysis revealed the major oligosaccharide moiety to be of the N-glycoside type. The carbohydrate chains were released from the iron-free form by digestion with peptide N-glycosidase F (PNGase F) in the presence of detergents such as SDS and-octylglucoside. After ethanol precipitation and fractionation on Bio-Gel P-6 and Bio-Gel P-2, the oligosaccharide was further purified on Mono-Q and desalted on Bio-Gel P-2. By 600-MHz1H-NMR spectroscopy, the primary structure of the major N-linked oligosaccharide component was established to be:
Abbreviations used HSmT
human seminal transferrin
- HSrT
human serum transferrin
- PNGase F
peptide-N4-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52), commonly known as peptide N-glycosidase F
- SDS-PAGE
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- GLC
gas-liquid chromatography
- FPLC
fast liquid protein chromatography
- EDTA
ethylenediaminetetraacetic acid, disodium salt
- PMFS
phenylmethylsulfonyl fluoride
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- Man, Gal
galactose
- Fuc
fucose 相似文献
11.
A facile method for introducing reactive sulphydryl groups intooligosaccharides was developed. 1-Amino-oligo-saccharides generatedfrom asparagine-linked glycans by peptide-N4(N-acetyl-ß-D-glucosaminyl)asparagine amidase (PNGase F) digestion were monitored by high-performanceanion-exchange chromatography with pulsed amperometric detectionand derivatized under optimal conditions with 2-iminothiolaneHC1.The resulting mercapto-butyramido oligosaccharides, which wereobtained in high yield, were alkylated with a fluorescent reagentand used to selectively assay for endoglycosidases that hydrolysedi-N-acetyl-chitobiose linkages. 1-amino-oligosaccharides fluorescent oligosaccharides 2-iminothiolane mercapto-butyramido oligosaccharides 相似文献
12.
Normal rat kidney cells, non-productively infected with the anaemia-inducing variant of Friend spleen focus-forming virus (F-SFFVA), were metabolically labelled with [2-3H]mannose. The primary translation product of the viral envelope gene (env), representing a glycoprotein with an apparent molecularM
r of 55 000 (gp55), was isolated from cell lysates by immunoaffinity chromatography and purified by preparative SDS/PAGE. Radiolabelled oligosaccharides, released from tryptic glycopeptides by treatment with endo--N-acetylglucosaminidase H, were characterized chromatographically, by enzymic digestion and by acetolysis. The results revealed that F-SFFVA gp55 obtained from this source carried predominantly oligomannose type sugar chains with five to nine mannoses. As a characteristic feature, glycans with seven to nine mannoses contained, in part, an additional glucose residue. Although the amount of glucosylated species found was higher in F-SFFVA gp55 (about 25% of total endo-H-sensitive oligosaccharides) than in gp55 of the corresponding polycythaemia-inducing variant (F-SFFVP, 16.3%), the overall glycosylation pattern of the F-SFFVA
env product closely resembled that of F-SFFVP gp55 [Strubeet al. (1988)J Biol Chem
263:3762–71]. Hence, our results demonstrate that the different intracellular processing and transport of the primary F-SFFVA
env product cannot be attributed to aberrant trimming of its oligomannose type glycans.Abbreviations endo H
endo--N-acetylglucosaminidase H fromStreptomyces griseus
-
env
envelope gene
- Env protein
translation product ofenv
- F-SFFV
Friend spleen focus-forming virus
- F-SFFVA
anaemia-inducing variant of F-SFFV
- F-SFFVP
polycythaemia-inducing variant of F-SFFV
- Hex
hexose
- NRK
normal rat kidney
- PNGase F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase F fromFlavobacterium meningosepticum 相似文献
13.
Summary This report describes the enzyme-catalyzed synthesis, characterization, and chromatographic separation of N6-(carboxymethyl)-L-lysine and N5-(carboxymethyl)-L-ornithine. The two N
-(carboxyalkyl)amino acids are formed via a reductive condensation between glyoxylate and the- or-amino groups of lysine and ornithine, respectively. Both reactions are catalyzed by the NADPH-dependent enzyme, N5-(carboxyethyl)ornithine synthase [EC 1.5.1.24], found in some strains of the lactic acid bacteriumLactococcus lactis subsp.lactis. 相似文献
14.
Alliinase (EC 4.4.1.4) catalyses the production of allicin (thio-2-propene-1-sulfinic acid S-allyl ester), a biologically active compound which is also responsible for the characteristic smell of garlic. It was demonstrated that alliinase which contains 5.5–6% of neutral sugars, gives clear PAS-staining, binds to Con A and can form a complex with garlic mannose-specific lectin (ASA). Evidence that the formation of such a complex is mediated by the interaction of the carbohydrate of the glycoprotein enzyme with the lectin was obtained from a radioligand assay which demonstrated the binding of alliinase to ASA and competitive inhibition of this binding by methyl -d-mannoside. ASA I was shown as the lectin mainly present in the complex with alliinase. The results of this study also demonstrate that alliinase is glycosylated at Asn146 in the sequence Asn146-Met147-Thr148.Abbreviations ASA
(Allium sativum agglutini). Garlic mannose specific lectin(s)
- PMSF
Phenyl methyl sulfonyl fluoride
- HPLC
High performance liquid chromatography
- SDS-PAGE
Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate
- PAS
Periodic acid-Schiff reagent stain
- CAPS
3-(cyclohexylamino)-1-propanesulfonic acid
- TFA
Trifluoracetic acid
- HEPES
N-2-HydroxyethylpiperazineN-2-ethanesulfonic acid
- Tricine
N-[2-hydroxyl-1,1-bis(hydroxymethyl)]-ethyl glycine
- PVDF
poly(vinylene difluoride) 相似文献
15.
The initial velocities of hydrolysis of nineteen glycopeptides by peptide: N-glycosidase F and A were determined. Substrates were prepared from bovine fetuin, hen ovalbumin, pineapple stem bromelain, bovine fibrin and taka-amylase. From these glycopeptides, several variants with regard to peptide and carbohydrate structure were prepared and derivatized with dabsyl chloride, dansyl chloride or activated resorufin. Tyrosine containing glycopeptides were also used without an additional chromophore. Enzymatic hydrolysis of glycopeptides was quantified by narrow bore, reversed phase HPLC with turnaround cycle times of down to 6 min, but usually 15 min.K
M values ranging from 30 to 64 µm and from 4 to 36 µm were found for N-glycosidase F and A, respectively. Relative velocities of hydrolysis of the different substrates by each enzyme varied considerably. Little, if any, similarity of the performance of N-glycosidase F and A with the different substrates was observed. The minimal carbohydrate structure released by peptide: N-glycosidase F was a di-N-acetylchitobiose. N-glycosidase A could release even a singleN-acetylglucosamine, albeit 3000 times slower than a di-N-acetylchitobiose or larger glycans. In general the structure of the intact glycan had little effect on activity, and with both enzymes the rate of hydrolysis appeared to be primarily governed by peptide structure and length. However, N-glycosidase F did not release glycans 1,3-fucosylated at the asparagine linkedN-acetylglucosamine irrespective of the presence of xylose in the substrate.Abbreviations CAMCys
S-carboxamidomethyl cystein
- CMCys
S-carboxymethyl cystein
- Fib, Fet, Ova, Taa and Brl
glycopeptides derived from bovine fibrin, fetuin, ovalbumin, taka-amylase A, and bromelain, respectively
- GlcNAc
N-acetylglucosamine
- PLA
phospholipase A2
- PNGase
peptide N-glycosidase
- RESOS
N-(Resorufin-4-carbonyl)piperidine-4-carboxylic acidN-hydroxysuccinimide ester 相似文献
16.
Jan B L Damm Johannis P Kamerling Gijs W K van Dedem Johannes F G Vliegenthart 《Glycoconjugate journal》1987,4(2):129-144
For the structural analysis of the carbohydrate chains ofN-,O-glycoproteins a straightforward strategy was developed based on the cleavage of theN-linked chains with immobilized peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase-F (PN-Gase-F) fromFlavobacterium meningosepticum, followed by alkaline borohydride treatment of the remainingO-glycoprotein material. This methodology was applied to the isolation of the Asn- and Ser-linked carbohydrate chains of human chorionic gonadotrophin. The structures of the isolated oligosaccharides were verified by 500-MHz1H-NMR spectroscopy. The Asn-linked sugar chains were shown to be: NeuAc2-3Gal1-4GlcNAc1-2Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man1-3]Man 1-4GlcNAc1-4[Fuc1-6]0-1GlcNAc and Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man 1-3]Man1-4GlcNAc1-4GlcNAc. Also some minor constituents occurred. The structures of the Ser-linked oligosaccharides were established in the form of their oligosaccharide-alditols as: NeuAc2-3Gal1-3[NeuAc2-6]GalNAc, NeuAc2-3Gal 1-3GalNAc and NeuAc2-3Gal1-3[NeuAc2-3Gal1-4GlcNAc1-6]GalNAc.Abbreviations hCG
human chorionic gonadotrophin
- hCG-
-subunit
- hCG-
-subunit
- ElA
enzyme immunoassay
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F (EC 3.5.1.52)
- SDS
sodium dodecyl sulphate
- GalNAc
N-acetylgalactosamine
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose 相似文献
17.
Gerd Reuter Roland Schauer Reginaldo Prioli Miercio E A Pereira 《Glycoconjugate journal》1987,4(4):339-348
In the culture supernatant ofTrypanosoma rangeli, strain El Salvador, a sialidase was present with an activity of 0.1 U/mg protein as determined with the 4-methylumbelliferyl glycoside of -N-acetylneuraminic acid as substrate. This enzyme was purified about 700-fold almost to homogeneity by gel chromatography on Sephadex G-100 and Blue Sepharose, and affinity chromatographies on 2-deoxy-2,3-didehydroneuraminic acid and horse submandibular gland mucin, both immobilized on Sepharose. The pH optimum is at 5.4–5.6, and the molecular weight was determined by gel chromatography, high performance liquid chromatography and sodium dodecyl sulphate gel electrophoresis to be 70 000. The substrate specificity of the enzyme is comparable to bacterial, viral and mammalian sialidases with cleavage rates for the following substrates in decreasing order:
N-acetylneuraminyl-(2–3)-lactose>
N-glycoloylneuraminy-(2–3)-lactose>
N-acetylneuraminyl-(2–6)-lactose >sialoglycoproteins>gangliosides>9-O-acetylated sialoglycoproteins.4-O-Acetylated derivatives are resistant towards the action of this sialidase. The enzyme activity can be inhibited by 2-deoxy-2,3-didehydro-N-acetylneuraminic acid, Hg2+ ions, andp-nitrophenyloxamic acid; it is not dependent on the presence of Ca2+ Mn2+ or Mg2+ ions.Abbreviations BSA
bovine serum albumin
- BSM
bovine submandibular gland mucin
- CMP
cytidine monophosphate
- EDIA
ethylenediaminetetraacetic acid
- ESM
equine submandibular gland mucin
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- HPLC
high performance liquid chromatography
- Lac
lactose
- MU-Neu5Ac
4-methylumbelliferyl glycoside of -N-acetylneuraminic acid
- Neu5Ac
N-acetylneuraminic acid
- Neu5Ac2en
2-deoxy-2,3-didehydro-N-acetylneuraminic acid
- Neu4Ac5Gc
N-glycoloyl-4-O-acetylneuraminic acid
- Neu2en
2-deoxy-2,3-didehydroneuraminic acid
- Neu5Gc
N-glycoloylneuraminic acid
- PMSF
phenylmethylsulfonyl fluoride
- PSM
pig submandibular gland mucin
- SDS
sodium dodecyl sulfate
- Tris
tris-(hydroxymethyl)aminomethane
Dedicated to Professor Dr. Heinz Mühlpfordt on the occasion of his 65th birthday. 相似文献
18.
Mutants of Pseudomonas aeruginosa deficient in the utilization of l-proline as the only carbon and nitrogen source have been found to be defective either in proline dehydrogenase activity or in both proline dehydrogenase and 1-pyrroline-5-carboxylate dehydrogenase activities of the bifunctional proline degradative enzyme. The latter type of mutants was unable to utilize l-ornithine, indicating that a single 1-pyrroline-5-carboxylate dehydrogenase activity is involved in the degradation of ornithine and proline. Proline dehydrogenase and 1-pyrroline-5-carboxylate dehydrogenase activities were strongly and coordinately induced by proline. It was excluded that 1-pyrroline-5-carboxylate acted as an inducer of the bifunctional enzyme and it was shown that the low level induction observed during growth on ornithine was due to the intracellular formation of proline. The formation of the proline degradative enzyme was shown to be subject to catabolite repression by citrate and nitrogen control.Abbreviations EMS
Ethylmethane sulfonate
- NG
N-methyl-N-nitro-N-nitrosoguanidine
- P
Minimal medium P
- Pro-DH
Proline dehydro-genase
- P5C
1-Pyrroline-5-carboxylate
- P5C-DH
1-Pyrroline-5-carboxylate dehydrogenase 相似文献
19.
Pediococcus halophilus possesses phosphoenolpyruvate:mannose phosphotransferase system (man:PTS) as a main glucose transporter. A man:PTS defective (man:PTSd) strain X-160 could, however, utilize glucose. A possible glucose-transport mechanism other than PTS was studied with the strain X-160 and its derivative, man:PTSd phosphofructokinase defective (PFK–) strain M-13. Glucose uptake by X-160 at pH 5.5 was inhibited by any of carbonylcyanide m-chlorophenylhydrazone, nigericin, N,N-dicyclohexylcarbodiimide, or iodoacetic acid. The double mutant M-13 could still transport glucose and accumulated intracellularly a large amount of hexose-phosphates (ca. 8 mM glucose 6-phosphate and ca. 2 mM fructose 6-phosphate). Protonophores also inhibited the glucose transport at pH 5.5, as determined by the amounts of accumulated hexose-phosphates (< 4 mM). These showed involvement of proton motive force (P) in the non-PTS glucose transport. It was concluded that the non-PTS glucose transporter operated in concert with hexokinase or glucokinase for the metabolism of glucose in the man:PTSd strain.Abbreviations BM
basal medium
- BM-G
basal medium containing glucose
- CM
complex medium
- man:PTS
phosphoenolpyruvate:mannose phosphotransferase system
- CCCP
carbonylcyanide m-chlorophenylhydrazone
- DCCD
N,N-dicyclohexyl carbodiimide
- P
proton motive force
- pH
transmembrane pH gradient
-
transmembrane electrical potential difference
- MNNG
N-methyl-N-nitro-N-nitrosoguanidine
- PIPES
piperazine-N,N-bis(-ethanesulfonic acid)
- MES
4-morpholineethanesulfonic acid
- G-6-P
glucose 6-phosphate
- F-6-P
fructose 6-phosphate
- FDP
fructose 1,6-bisphosphate
- EMP
Embden-Meyerhof-Parnas pathway
- PFK
phosphofructokinase
- GK
glucokinase
- HK
hexokinase
- IAA
iodoacetic acid
- IIman
enzyme II component of man:PTS 相似文献
20.
William R. Strohl Thomas M. Schmidt Victor A. Vinci John M. Larkin 《Archives of microbiology》1986,145(1):71-75
Seven strains of bacteria belonging to the Beggiatoa-Vitreoscilla group were studied for their respiratory activity and for the presence of electron transport conponents. All strains tested oxidized [1-14C] and [2-14C] acetate to 14CO2 at relatively high rates. All strains tested were N,N,N,N-tetramethylphenylenediamine (TMPD)-oxidase positive and contained spectra representing a-type and carbon monoxide-binding cytochromes. Most of the strains also contained spectra representing c-type and b-type cytochromes. Beggiatoa alba B18LD contained b-type, a-type, c-type and CO-binding cytochromes, the latter two being located in the 144,000 x g soluble fraction. B. alba also contained ubiquinone-8 as its only detectable quinone.Non-standard abbreviations BSS
basal salts solution
- BH
Beggiatoa heterotrophic medium
- BSO
Beggiatoa sulfide oxidation medium
- TMPD
N,N,N,N-tetramethylphenylenediamine
- Q8
ubiquinone-8 相似文献