scholarly journals The biosynthesis of valine from isobutyrate by Peptostreptococcus elsdenii and Bacteroides ruminicola

1971 ◽  
Vol 121 (3) ◽  
pp. 431-437 ◽  
Author(s):  
Milton J. Allison ◽  
J. L. Peel
Keyword(s):  
Molecular Hydrogen ◽  
Photosynthetic Bacteria ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Acetyl Phosphate ◽  
Acid Fraction ◽  
Benzyl Viologen ◽  
Electron Carrier ◽  
Thiamin Pyrophosphate ◽  
Reductive Carboxylation

1. Growing cultures of Peptostreptococcus elsdenii and Bacteroides ruminicola incorporate 14C from [1-14C]isobutyrate into the valine of cell protein. With P. elsdenii some of the 14C is also incorporated into leucine. 2. Crude cell-free extracts of both organisms in the presence of glutamine, carbon dioxide and suitable sources of energy and electrons incorporate 14C from [1-14C]isobutyrate into valine but not into leucine. 3. With extracts of P. elsdenii treated with DEAE-cellulose the reaction is dependent on ATP, CoA, thiamin pyrophosphate, molecular hydrogen and a low-potential electron carrier (ferredoxin, flavodoxin or benzyl viologen). 4. The same extracts incorporate 14C from NaH14CO3 into valine in the presence of isobutyrate plus ATP, CoA, glutamine and ferredoxin; isobutyryl-CoA or isobutyryl phosphate plus CoA will replace the isobutyrate plus CoA and ATP. With acetyl phosphate in place of isobutyryl phosphate, 14C is incorporated into alanine. With isovalerate or 2-methylbutyrate in place of isobutyrate, 14C is incorporated into leucine and isoleucine respectively. 5. When carrier 2-oxoisovalerate is added to the carboxylating system 14C from [1-14C]isobutyrate passes into the oxo acid fraction. 6. It is concluded that these two organisms form valine from isobutyrate by the sequence isobutyrate→isobutyryl-CoA→2-oxoisovalerate→valine and that the reductive carboxylation of isobutyrate is catalysed by a system similar to the pyruvate synthetase of clostridia and photosynthetic bacteria.

Pyruvate oxidation by the Reiter strain of Treponema phagedenis

1982 ◽  
Vol 152 (3) ◽  
pp. 1060-1065
Author(s):  
H A George ◽  
R M Smibert
Keyword(s):  
Gel Filtration ◽  
Deae Cellulose ◽  
Treponema Denticola ◽  
Benzyl Viologen ◽  
Iron Chelate ◽  
Electron Carrier ◽  
Pyruvate Oxidation ◽  
Crude Extracts ◽  
Pyruvate:Ferredoxin Oxidoreductase ◽  
Triton X

Spectrophotometric assays of pyruvate oxidation catalyzed by extracts of the Reiter strain of Treponema phagedenis indicated that viologen dyes, flavin nucleotides, and a ferric iron chelate, but not pyridine nucleotides, were utilized as electron acceptors. Benzyl viologen-linked activity partially sedimented during ultracentrifugation and appeared similar to clostridial pyruvate:ferredoxin oxidoreductase with respect to the spectral properties of the enzyme chromophore. Electron carrier activity in treponemal extracts was quantitated by a metronidazole-linked assay in which the oxidation of pyruvate by carrier-depleted extracts led to the reduction of electron carrier in the crude extracts which then reduced metronidazole. The rate of metronidazole reduction was proportional to the amount of electron carrier present in the assay. Electron carrier activity in Triton X-100-solubilized, crude extracts partially purified by DEAE-cellulose chromatography and gel filtration was attributed to a protein possessing the spectral and physical properties of a ferredoxin. A similar protein appeared to be present in extracts of Treponema denticola ST10.

scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132 ◽  
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Abstract Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

scholarly journals Production of molecular hydrogen by photosynthetic bacteria with raw starch.

1985 ◽  
Vol 49 (11) ◽  
pp. 3339-3341 ◽  
Author(s):  
Lerluck BURANAKARL ◽  
FAN Cheng-Ying ◽  
Kazutoshi ITO ◽  
Kazuo IZAKI ◽  
Hajime TAKAHASHI
Keyword(s):  
Molecular Hydrogen ◽  
Photosynthetic Bacteria ◽  
Raw Starch

Production of Molecular Hydrogen by Photosynthetic Bacteria with Raw Starch

1985 ◽  
Vol 49 (11) ◽  
pp. 3339-3341
Author(s):  
Lerluck Buranakarl ◽  
Fan Cheng-Ying ◽  
Kazutoshi Ito ◽  
Kazuo Izaki ◽  
Hajime Takahashi
Keyword(s):  
Molecular Hydrogen ◽  
Photosynthetic Bacteria ◽  
Raw Starch

scholarly journals Immunoassay of bovine and human low-density-lipoprotein receptors using monoclonal antibodies

1986 ◽  
Vol 238 (2) ◽  
pp. 405-410 ◽  
Author(s):  
B L Knight ◽  
S Preyer ◽  
A K Soutar
Keyword(s):  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Deae Cellulose ◽  
Density Lipoprotein ◽  
Receptor Protein ◽  
Cell Protein ◽  
Human Skin Fibroblast ◽  
Low Density ◽  
Normal Human Skin ◽  
Routine Assay

Two methods are described for the assay of low-density-lipoprotein (LDL) receptor protein based on the binding of receptor to microtitre plate wells coated with a specific monoclonal antibody or with LDL, followed by detection with radioactive antibody that recognizes a different part of the molecule. The two-antibody procedure detected approx. 2 ng of pure bovine receptor at twice background, and there was a linear relationship on a double-logarithm plot between radioactive antibody bound and the amount of receptor added, up to at least 500 ng of receptor protein per well. The procedure using immobilized LDL was less sensitive and the binding of receptor was inhibited by low concentrations of NaCl, which restricted its usefulness for routine assay of tissue extracts. LDL receptor protein could be readily assayed using the two-antibody procedure in normal human skin fibroblast extracts prepared by bulk-elution from small columns of DEAE-cellulose followed by rapid desalting. No radioactive antibody bound with extracts of cells from a receptor-negative familial hypercholesterolaemic subject. The LDL receptor content of normal fibroblasts preincubated with lipoprotein-deficient serum was estimated, using bovine receptor as standard, to be approx. 60 ng of receptor protein/mg of cell protein.

scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

Sign in / Sign up

Export Citation Format

Share Document