5-Methyltetrahydrofolic acid and other folate derivatives in germinating pea seedlings

1968 ◽  
Vol 46 (12) ◽  
pp. 1533-1536 ◽  
Author(s):  
A. J. Roos ◽  
A. M. Spronk ◽  
E. A. Cossins
Keyword(s):  
Folic Acid ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Methyl Derivative ◽  
Streptococcus Faecalis ◽  
Feeding Experiments ◽  
Pea Seedlings ◽  
Young Leaves

The folate derivatives present in germinating pea seedlings were isolated by chromatography on DEAE-cellulose and assayed with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae. The major folate derivative in the cotyledons, developing embryos, and young leaves was identified as 5-methyltetrahydrofolate. Smaller amounts of 10-formyltetrahydrofolate were also present in these tissues. Synthesis of these derivatives in the cotyledons was inhibited by aminopterin. Feeding experiments showed that the 5-methyl derivative was rapidly synthesized from folic acid-2-14C.

Composition of Pteroylpolyglutamates (Conjugated Folates) in Guinea-Pig Liver and Their Formation from Folic Acid

1972 ◽  
Vol 43 (6) ◽  
pp. 799-813 ◽  
Author(s):  
R. Corrocher ◽  
B. K. Bhuyan ◽  
A. V. Hoffbrand
Keyword(s):  
Folic Acid ◽  
Guinea Pig ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Microbiological Assay ◽  
Pig Liver ◽  
Ion Exchange Column ◽  
Pteroylglutamic Acid ◽  
Guinea Pig Liver ◽  
Liver Folate

1. The composition of guinea-pig liver folates and the biochemical route of formation of liver folates from injected tritium-labelled pteroylglutamic acid (folic acid) have been studied. 2. Endogenous folate was measured by microbiological assay with Lactobacillus casei and Streptococcus faecalis, with and without deconjugation of whole liver pteroylpolyglutamates (conjugated folates). Individual folate compounds were identified by microbiological assay after fractionation of liver folates by DEAE cellulose ion-exchange column chromatography. 3. Liver folate in the guinea-pig consists of about 84–87% reduced pteroylpolyglutamates with more than three glutamate moieties/molecule, about 12–15% reduced pteroyltriglutamates, about 1% reduced pteroyldiglutamates and only traces of reduced pteroylmonoglutamates. 4. About 53% of the liver folate consists of methylated derivatives. 5. Injected pteroylglutamic acid was first rapidly reduced and formylated or methylated. Glutamate moieties were then added, probably singly, to form di-, tri- and poly-glutamates. This was a relatively slow process with a hold-up at the triglutamate stage. 6. The proportion of the labelled pteroylglutamic acid in the polyglutamate form approximated to the proportion of endogenous folates in this form after 3–4 days. 7. The amount of radioactive folate in the liver increased progressively from 1 to 84 h after injection of a standard amount of radioactive pteroylglutamic acid.

Effect of excision and benzimidazole treatment on folate content of wheat leaves and wheat leaf chloroplasts

1970 ◽  
Vol 48 (10) ◽  
pp. 1091-1095 ◽  
Author(s):  
W. K. Kim
Keyword(s):  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Streptococcus Faecalis ◽  
Crude Extracts ◽  
Wheat Leaf ◽  
Wheat Leaves ◽  
Peak Fraction

Crude extracts were prepared from freshly harvested wheat leaves, and from leaves floated on water or on solutions of benzimidazole. Extracts were also prepared from chloroplasts isolated with non-aqueous solvents from similar leaves. All extracts were fractionated on DEAE-cellulose columns. Folate content was determined with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae as assay organisms.Total folates increased in leaves after excision whether benzimidazole was present or not. The increase was mainly due to an increase of conjugates of 5-methyltetrahydrofolate. Similar folate drifts were observed in chloroplasts isolated from these samples. Peak fraction analyses indicated that methylated folate conjugates were converted to 5-methyltetrahydrofolate, and formylated folates disappeared in leaves floated on water. Benzimidazole treatment arrested the degradation of conjugated folates, except that of a formylated folate found only in chloroplasts.

Folate derivatives in ungerminated and germinated uredospores of wheat stem rust

1970 ◽  
Vol 48 (9) ◽  
pp. 1617-1623
Author(s):  
A. O. Jackson ◽  
D. J. Samborski ◽  
R. Rohringer ◽  
W. K. Kim
Keyword(s):  
Growth Factors ◽  
Glutamic Acid ◽  
Detailed Analysis ◽  
Stem Rust ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Wheat Stem Rust ◽  
Streptococcus Faecalis ◽  
Peak Fraction ◽  
Methyl Tetrahydrofolate

Extracts of ungerminated uredospores and of uredospores germinated for 6 h and 12 h were assayed for folates with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae. These organisms did not respond to the extracts without conjugase treatment, indicating that most of the folates were present in conjugated form with more than three glutamic acid moieties per molecule. During the 6-h and 12-h germination periods the content of growth factors for L. casei declined to 70.9% and 46.0% of initial levels. In this same period, the content of growth factors for S. faecalis declined to 54.5% and 15.2% of initial levels, indicating an increase in the proportion of methylated folates during uredospore germination.This trend was confirmed in a detailed analysis of folate components after fractionation of the extracts on DEAE-cellulose columns. The folate profiles consisted of five peaks. Two peak fractions present in profiles from ungerminated spores contained mostly formylated folates and were greatly reduced or absent in profiles from germinated spores. A peak fraction composed of a 5-methyl-tetrahydrofolate conjugate was not observed in profiles of ungerminated spores but was predominant in those from spores germinated for 12 h.

Rapid Transfer of Folic Acid from Blood to Bile in Man, and its Conversion into Folate Coenzymes and into a Pteroylglutamate with Little Biological Activity

1974 ◽  
Vol 46 (6) ◽  
pp. 729-741 ◽  
Author(s):  
A. Lavoie ◽  
B. A. Cooper
Keyword(s):  
Biological Activity ◽  
Folic Acid ◽  
Rose Bengal ◽  
Lactobacillus Casei ◽  
Streptococcus Faecalis ◽  
Human Bile ◽  
Methylene Tetrahydrofolate ◽  
Special Modification ◽  
A Chain ◽  
Rapid Transfer

1. [3H]Folic acid infused intravenously into patients with biliary fistulae appeared promptly in bile, coincident with 131I-labelled Rose Bengal injected simultaneously. The radioactivity was distributed among several fractions of biological folates present in bile and was associated with folic acid and with an unidentified folate which chromatographed on DEAE-Sephadex close to 10-formyltetrahydrofolyl monoglutamate. 2. Based on affinity for DEAE-Sephadex and support of growth of Lactobacillus casei, Streptococcus faecalis and Pediococcus cerevisiae, we have tentatively identified some of the folates of human bile as 10-formyltetrahydrofolate, 10-formylfolate, 5-formyltetrahydrofolate, 5-methyltetrahydrofolate and tetrahydrofolate or 5,10-methylene tetrahydrofolate. After infusion of folic acid, the formyltetrahydrofolates increased more rapidly than did 5-methyltetrahydrofolate. 3. The unidentified radioactive folate contained both the pteridine and p-aminobenzoate portions of folate. It appeared not to support growth of the test microorganisms and not to be bound to a protein or a chain of γ-glutamates. It was present in the bile of a dog injected with [3H]folic acid but was absent from extracts of liver. This material may be a transport form of folate or a special modification imposed on folic acid during transport across the liver.

Folate derivatives in healthy and rust-infected primary leaves of wheat

1969 ◽  
Vol 47 (12) ◽  
pp. 1161-1169 ◽  
Author(s):  
R. Rohringer ◽  
W. K. Kim ◽  
D. J. Samborski
Keyword(s):  
Growth Factors ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Resistant Line ◽  
Streptococcus Faecalis ◽  
Susceptible Line ◽  
Crude Extracts ◽  
Wheat Leaves ◽  
Chicken Pancreas

Six days after inoculation, crude extracts were prepared from healthy and rust-infected wheat leaves of a susceptible line and from healthy and rust-infected wheat leaves of a resistant line. Extracts from rust-infected leaves of both lines contained 50% more folates, active as growth factors for Lactobacillus casei, than did those from healthy leaves. Rust-infected leaves did not differ from healthy leaves in their content of folates active as growth factors for Streptococcus faecalis and Pediococcus cerevisiae.All extracts were fractionated on DEAE-cellulose columns. The eluate fractions were treated with chicken pancreas conjugase and assayed with L. casei. All extracts yielded similar folate profiles consisting of five major and three minor peaks. Conjugase treatment and differential assay of the peak fractions with L. casei, S. faecalis, and P. cerevisiae indicated that the eluates contained folates methylated at N-5 and folates formylated at N-10 in various states of oxidation and conjugation. The eluates also contained a compound believed to be 5-formyltetrahydropteroylglutamate, small amounts of pteroylglutamate, traces of tetrahydropteroylglutamate, and several unidentified folates.The increase of folate levels in rust-infected leaves was due almost entirely to increases of 5-methyl-tetrahydropteroylgiutamate and its conjugates. The folate composition of resistant-reacting leaves did not differ appreciably from that of susceptible-reacting leaves.Radioactivity was not incorporated into either 5-methyltetrahydropteroyltriglutamate or into methionine when 5-[methyl-14C]-tetrahydropteroylglutamate was fed to wheat leaves.

scholarly journals Pteroylglutamate derivatives in Pisum sativum L. Biosynthesis of cotyledonary tetrahydropteroylglutamates during germination

1971 ◽  
Vol 125 (1) ◽  
pp. 17-26 ◽  
Author(s):  
A. J. Roos ◽  
E. A. Cossins
Keyword(s):  
Pisum Sativum ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Cellulose Synthesis ◽  
Feeding Experiments ◽  
Tissue Extracts ◽  
Pisum Sativum L ◽  
Pteroylglutamic Acid ◽  
Derivatives Of ◽  
Endogenous Enzymes

1. The concentrations of pteroylglutamate derivatives in the cotyledons of pea (Pisum sativum L.) seedlings were determined by microbiological assay by using Lactobacillus casei (A.T.C.C. 7469), Streptococcus faecalis (A.T.C.C. 8043) and Pediococcus cerevisiae (A.T.C.C. 8081). During germination the pteroylglutamate content of the cotyledons increased rapidly from 0.2μg/g dry wt. to 4.0μg/g dry wt., the maximum values being reached approx. 120h after imbibition. 2. Approx. 50% of the pteroylglutamate pool of 3-day-old seedlings was accounted for by highly conjugated derivatives. The concentrations of such derivatives were greatest when precautions were taken to inactivate endogenous enzymes before extraction of the tissues. 3. Individual derivatives present in the tissue extracts were separated by chromatography on DEAE-cellulose. Synthesis of the major derivative, 5-methyltetrahydropteroylmonoglutamate, was inhibited by administration of 0.1mm-aminopterin and -amethopterin solutions during imbibition. Under these conditions pteroylglutamic acid accumulated in the tissues. 4. Feeding experiments employing [2-14C]pteroylglutamic acid and 5[14C]-methyltetrahydro-pteroylmonoglutamic acid revealed that both compounds were incorporated into conjugated and unconjugated derivatives of the pteroylglutamate pool.

scholarly journals Folic acid content of beetroot leaf and root by different growing stages and genotypes

2019 ◽  
pp. 115-119
Author(s):  
Tímea Rubóczki ◽  
Mária Takácsné Hájos
Keyword(s):  
Folic Acid ◽  
Acid Content ◽  
Dry Matter ◽  
Developmental Stages ◽  
Matter Content ◽  
Nitrate Content ◽  
Dry Matter Content ◽  
Randomized Field Experiment ◽  
Sowing Dates ◽  
Young Leaves

An increasing interest has been observed of beetroot leaf as a salad component due to recent studies focusing on their nutritional value. The randomized field experiment was carried out on lowland chernozem soil with 6 varieties, 3 replications and 2 sowing dates. Sampling was performed on 23 of August 2018 at the stage of 30 and 50 days of vegetation, where leaf (30 and 50 days) and root (50 days) were collected. Total dry matter, folic acid and nitrate content were evaluated. The results of this investigation show that higher total dry matter content was measured in the root (8.47–10.30%) compared to the leaf in both developmental stages (6.47–9.20%). Nevertheless, higher folic acid content was found in the young leaves of 30 and 50 days of development (58.77–113.86 µg 100g-1). Among the examined varieties, Bonel has presented great amount of folic acid not only in the leaves (99.35–113.61 µg 100g-1), but also in the root (89.99 µg 100g-1). Finally, lower nitrate content was found in Libero (316.16 mg kg-1) at 30 days and in Akela (340.41 mg kg-1) at 50 days of development. Thereby, fresh consumption of beetroot leaves are highly recommended.

Folic acid compounds in romaine lettuce

1977 ◽  
Vol 55 (8) ◽  
pp. 865-868 ◽  
Author(s):  
K. K. Batra ◽  
J. R. Wagner ◽  
E. L. R. Stokstad
Keyword(s):  
Folic Acid ◽  
Specific Activity ◽  
Deae Cellulose ◽  
High Specific Activity ◽  
Microbiological Assay ◽  
Romaine Lettuce ◽  
Positive Identification

The composition of folate coenzymes in romaine lettuce was studied. Lettuce extract was purified on QAE-Sephadex A-25 and folate compounds were separated into a monoglutamate fraction and a polyglutamate fraction by chromatography on Sephadex G-15. Both the mono- and poly-glutamate fractions were resolved on DEAE-cellulose. Positive identification of DEAE peaks was made by further cochromatography with high specific activity radioactive marker folate compounds and with differential microbiological assay. The distribution of folate compounds in lettuce is as follows: 32% 5-CH3-H4PteGlu; 1% 5-CHO-H4PteGlu; 3% 5-CHO-H4PteGlu4; 9% 5-CH3-H4PteGlu4; 13% 5-CHO-H4PteGlu5; and 31% 5-CH3-H4PteGlu5.

scholarly journals Localization and interconversion of tetrahydropteroylglutamates in isolated pea mitochondria

1972 ◽  
Vol 128 (1) ◽  
pp. 29-40 ◽  
Author(s):  
M. T. Clandinin ◽  
E. A. Cossins
Keyword(s):  
Amino Acids ◽  
Free Amino ◽  
Lactobacillus Casei ◽  
Sucrose Density Gradient ◽  
Mitochondrial Fraction ◽  
C1 Metabolism ◽  
Isolated Mitochondria ◽  
Feeding Experiments ◽  
Pteroylglutamic Acid ◽  
Derivatives Of

1. Mitochondria were extracted from 4-day-old pea cotyledons and purified on a sucrose density gradient. 2. Microbiological assay of the purified mitochondrial fraction with Lactobacillus casei (A.T.C.C. 7469), Streptococcus faecalis (A.T.C.C. 8043) and Pediococcus cerevisiae (A.T.C.C. 8081) revealed a discrete pool of conjugated and unconjugated derivatives of tetrahydropteroylglutamic acid. 3. Solubilization and chromatographic studies of the mitochondrial fraction demonstrated the presence of formylated and methylated derivatives, 10-formyltetrahydropteroylmonoglutamic acid, 5-formyltetrahydropteroylmonoglutamic acid and 5-formyltetrahydropteroyldiglutamic acid being the major derivatives present. 4. The principal mitochondrial pteroylglutamates were labelled when dry seeds were allowed to imbibe [2-14C]pteroylglutamic acid and 5-[methyl-14C]-methyltetrahydropteroylmonoglutamic acid. 5. The ability of isolated mitochondria to catalyse oxidation and reduction of tetrahydropteroylglutamic acid derivatives was demonstrated in feeding experiments in which [14C]formaldehyde, [3-14C]serine, sodium [14C]formate, 5-[methyl-14C]methyltetrahydropteroylmonoglutamic acid or [2-14C]-glycine served as C1 donor. In addition,14C was incorporated into free amino acids related to C1 metabolism.

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