scholarly journals Modification of isolation methods and physico-biochemical properties of preparations of fungal oxidoreductases

2020 ◽  
Vol 11 (2) ◽  
pp. 310-314
Author(s):  
O. V. Fedotov ◽  
Z. L. Usikova
Keyword(s):  
Amino Acids ◽  
Lentinula Edodes ◽  
Gel Filtration ◽  
Amino Acid Content ◽  
Toxicity Testing ◽  
Culture Fluid ◽  
Biochemical Properties ◽  
Individual Characteristics ◽  
Intracellular Location ◽  
Enzyme Preparations

The results of the modification of methods for producing enzymatic preparations (EP) of peroxidases and catalases of extra- and intracellular finding from fungal cultures are presented. Strains of Flammulina velutipes F-vv, Lentinula edodes 523 and Pleurotus ostreatus P-01 were used as producers of oxidoreductases. The producers were cultured in glucose-peptone medium, modified for each strain. Protein fractionation was carried out by leaching with ammonium sulfate at a saturation of 40–70% for peroxidases and 80% for catalases. The obtained solutions of protein fractions were additionally subjected to purification by dialysis, gel filtration on Molselect G-50 and G-75 granules, and also freeze drying. The yield of enzymatic preparations per unit mass of mycelium and the volume of culture fluid were calculated. The individual characteristics of EP – enzymatic activity, the mass percentage of protein and associated amino acids, the ratio of the latter in groups depending on the nature of the radicals (amphotericity) of protein molecules are established. It was proved that the amino acid content in the proteins of fungal EP catalases and peroxidases indicates their acidic nature and this is confirmed by the pH values of aqueous solutions. Examination and toxicity testing of enzymatic preparations were carried out in certified laboratories, which confirmed their characteristics and compliance with safety requirements. The therapeutic properties of amino acids that are part of proteins or are in a free state in enzyme preparations are analyzed. In this way, the methods have been developed for producing enzymatic preparations of peroxidases and catalases of extra- and intracellular location, which allow new antioxidant enzymes with individual properties to be obtained, and, as a result, bring prospects for use in various industries and scientific research.

scholarly journals Modification of isolation methods and physico-biochemical properties of preparations of fungal oxidoreductases

2020 ◽  
Vol 11 (2) ◽  
pp. 310-314
Author(s):  
O. V. Fedotov ◽  
Z. L. Usikova
Keyword(s):  
Amino Acids ◽  
Lentinula Edodes ◽  
Gel Filtration ◽  
Amino Acid Content ◽  
Toxicity Testing ◽  
Culture Fluid ◽  
Biochemical Properties ◽  
Individual Characteristics ◽  
Intracellular Location ◽  
Enzyme Preparations

The results of the modification of methods for producing enzymatic preparations (EP) of peroxidases and catalases of extra- and intracellular finding from fungal cultures are presented. Strains of Flammulina velutipes F-vv, Lentinula edodes 523 and Pleurotus ostreatus P-01 were used as producers of oxidoreductases. The producers were cultured in glucose-peptone medium, modified for each strain. Protein fractionation was carried out by leaching with ammonium sulfate at a saturation of 40–70% for peroxidases and 80% for catalases. The obtained solutions of protein fractions were additionally subjected to purification by dialysis, gel filtration on Molselect G-50 and G-75 granules, and also freeze drying. The yield of enzymatic preparations per unit mass of mycelium and the volume of culture fluid were calculated. The individual characteristics of EP – enzymatic activity, the mass percentage of protein and associated amino acids, the ratio of the latter in groups depending on the nature of the radicals (amphotericity) of protein molecules are established. It was proved that the amino acid content in the proteins of fungal EP catalases and peroxidases indicates their acidic nature and this is confirmed by the pH values of aqueous solutions. Examination and toxicity testing of enzymatic preparations were carried out in certified laboratories, which confirmed their characteristics and compliance with safety requirements. The therapeutic properties of amino acids that are part of proteins or are in a free state in enzyme preparations are analyzed. In this way, the methods have been developed for producing enzymatic preparations of peroxidases and catalases of extra- and intracellular location, which allow new antioxidant enzymes with individual properties to be obtained, and, as a result, bring prospects for use in various industries and scientific research.

Purification of an exo-1,3-β-glucanase from Candida utilis

1976 ◽  
Vol 54 (11) ◽  
pp. 927-934 ◽  
Author(s):  
T. G. Villa ◽  
V. Notario ◽  
T. Benítez ◽  
J. R. Villanueva
Keyword(s):  
Amino Acids ◽  
Heavy Metal ◽  
Cell Wall ◽  
Heavy Metal Ions ◽  
Candida Utilis ◽  
Negative Charge ◽  
Hydrolysis Product ◽  
Culture Fluid ◽  
Biochemical Properties ◽  
Acidic Amino Acids

An exo-1,3-β-glucanase (EC 3.2.1.—) has been purified from the culture fluid of the yeast Candida utilis, and its biochemical properties have been studied. The amino acid analysis revealed a high content of acidic amino acids. The purified enzyme had 20% carbohydrate and a net negative charge showing higher affinity for laminarin than for p-nitrophenyl-β-D-glucopyranoside and yeast cell-wall 1,3-β-glucans. In addition, the enzyme hydrolyzed the substrates starting from the nonreducing ends, releasing glucose as the exclusive hydrolysis product. The enzyme activity was strongly inhibited by lactones and also by some heavy-metal ions.

scholarly journals Evidence for Production of a New Lantibiotic (Butyrivibriocin OR79A) by the Ruminal Anaerobe Butyrivibrio fibrisolvensOR79: Characterization of the Structural Gene Encoding Butyrivibriocin OR79A

1999 ◽  
Vol 65 (5) ◽  
pp. 2128-2135 ◽  
Author(s):  
M. L. Kalmokoff ◽  
D. Lu ◽  
M. F. Whitford ◽  
R. M. Teather
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Gel Filtration ◽  
Culture Fluid ◽  
Northern Blotting ◽  
Coding Region ◽  
Gene Encoding ◽  
Butyrivibrio Fibrisolvens ◽  
Reading Frame ◽  
Significant Homology

ABSTRACT The ruminal anaerobe Butyrivibrio fibrisolvens OR79 produces a bacteriocin-like activity demonstrating a very broad spectrum of activity. An inhibitor was isolated from spent culture fluid by a combination of ammonium sulfate and acidic precipitations, reverse-phase chromatography, and high-resolution gel filtration. N-terminal analysis of the isolated inhibitor yielded a 15-amino-acid sequence (G-N/Q-G/P-V-I-L-X-I-X-H-E-X-S-M-N). Two different amino acid residues were detected in the second and third positions from the N terminus, indicating the presence of two distinct peptides. A gene with significant homology to one combination of the determined N-terminal sequence was cloned, and expression of the gene was confirmed by Northern blotting. The gene (bvi79A) encoded a prepeptide of 47 amino acids and a mature peptide, butyrivibriocin OR79A, of 25 amino acids. Significant sequence homology was found between this peptide and previously reported lantibiotics containing the double-glycine leader peptidase processing site. Immediately downstream of bvi79Awas a second, partial open reading frame encoding a peptide with significant homology to proteins which are believed to be involved in the synthesis of lanthionine residues. These findings indicate that the isolated inhibitory peptides represent new lantibiotics. Results from both total and N-terminal amino acid sequencing indicated that the second peptide was identical to butyrivibriocin OR79A except for amino acid substitutions in positions 2 and 3 of the mature lantibiotic. Only a single coding region was detected when restriction enzyme digests of total DNA were probed either with an oligonucleotide based on the 5′ region of bvi79A or with degenerate oligonucleotides based on the predicted sequence of the second peptide.

scholarly journals The chemistry of the collagen cross-links. Purification and characterization of cross-linked polymeric peptide material from mature collagen containing unknown amino acids

1980 ◽  
Vol 185 (2) ◽  
pp. 373-381 ◽  
Author(s):  
N D Light ◽  
A J Bailey
Keyword(s):  
Amino Acids ◽  
Type I Collagen ◽  
Gel Filtration ◽  
Amino Acid Content ◽  
Exchange Chromatography ◽  
Type I ◽  
Polymeric Form ◽  
Cross Links ◽  
Helical Region

A polymeric form of the alpha 1-chain C-terminal peptide alpha 1 CB6 (poly-alpha 1 CB6) was purified from CNBr digests of insoluble bovine tendon type-I-collagen by gel filtration and ion-exchage chromatography. The purified material had a molecular weight of 1.5 × 10(6)-5 × 10(6) on gel filtration and an amino acid content virtually identical with that of monomeric peptide alpha 1 CB6. The material could be adsorbed on affinity gels containing immobilized anti-(alpha 1 CB6-peptide non-helical region) antibodies and was an inhibitor of haemagglutination by the same antibodies of alpha 1 CB6-peptide-coated sheep erythrocytes. Periodate treatment of the material had no effect. Alkali hydrolysates were shown to contain two unknown amino acids, which were purified by gel filtration and ion-exchange chromatography in volatile buffers and are believed to be components of the mature cross-link of collagen.

scholarly journals РОЗРОБКА СПОСОБІВ ОТРИМАННЯ І АНАЛІЗ ФЕРМЕНТНИХ ПРЕПАРАТІВ ПЕРОКСИДАЗ ТА КАТАЛАЗ ДЕЯКИХ ВИДІВ БАЗИДІОМІЦЕТІВ

2013 ◽  
Vol 3 (01) ◽  
pp. 113 ◽  
Author(s):  
O. V. Fedotov ◽  
T. E. Voloshko
Keyword(s):  
Freeze Drying ◽  
Enzyme Preparation ◽  
Gel Filtration ◽  
Science Research ◽  
Individual Characteristics ◽  
Salting Out ◽  
Enzyme Preparations ◽  
Agrocybe Cylindracea ◽  
Activity Of Enzymes ◽  
The Individual

<p>A method for obtaining of enzyme preparations of enzyme preparations (EP) of peroxidases and catalases fungal extracellular and inracellular origin from cultures of Basidiomycetes was developed. The strains <em>Flammulina velutipes</em> F-vv, <em>Agrocybe cylindracea</em> 167; <em>Fistulina hepatica</em> Fh-08 and <em>Pleurotus ostreatus</em> P-208 and P-01 were used as producers of oxidoreductases. Strains were grown on modified glucose-peptone media. Fractionation was carried out by salting out the enzymes with ammonium sulfate at 40-70% saturation of peroxidases and 80% of saturation - for catalase. These solutions protein fractions was further purified by dialysis and gel filtration on Molselekt granules G-50 and G-75. The enzyme solution was subjected to freeze-drying. The individual characteristics of the enzyme preparations were found.  The individual characteristics of the enzyme preparations are the activity of enzymes, the protein content and amino-acid composition of enzyme preparations. It was established that strain <em>F. velutipes</em> F-vv was an active producer of intracellular and strain of <em>A. cylindracea</em> 167 was an active producer of extracellular peroxidase. The strains of <em>P. ostreatus</em> P-01 and P-208 were the active producers of extracellular catalase, and the strains of F. hepatica Fh-08 were active producers of intracellular catalase. The developed methods for producing of enzymes catalase and peroxidase preparations of extra-and intracellular origin provided new antioxidant enzymes, which have their own properties and application prospects in various sectors of industry and science research.</p> <p><em>Key words: Basidiomycetes, peroxidases, catalases, enzyme preparation.</em></p>

Screening of amino acid constituents from date palm fruits

2016 ◽  
Vol 5 (10) ◽  
pp. 4972
Author(s):  
Lata Birlangi
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
United Arab Emirates ◽  
Nutritional Value ◽  
Date Palm ◽  
Amino Acid Content ◽  
Essential Amino Acids ◽  
Cultivated Plants ◽  
Palm Fruits ◽  
Social Part

The date palm (Phoenix dactylifera L.) is one of mankind’s oldest cultivated plants. The fruit of the date palm is an important crop of the hot arid and semi-arid regions of the world. It has always played a genuine economic and social part in the lives of the people of these areas. The present objective in examining the amino acid content of different varieties of date palm fruits from Middle-East region; is to determine whether its protein could effectively supplement the nutritional value and it is also aimed in finding which variety is rich in number of amino acids. The phytochemical screening revealed the presence of eight essential amino acids and five non-essential amino acids in the date fruits. Among all the date fruit varieties taken as samples for the study, Dabbas cultivar of United Arab Emirates found to exhibit eight types of amino acids which includes five as non-essential ones. Total of thirteen amino acids were detected in the seven date cultivars. Determination of amino acid can serve as a guide to the possible nutritional value.

scholarly journals Putative imbalanced amino acid metabolism in rainbow trout long term fed a plant-based diet as revealed by 1H-NMR metabolomics

2021 ◽  
Vol 10 ◽  
Author(s):  
Catherine Deborde ◽  
Blandine Madji Hounoum ◽  
Annick Moing ◽  
Mickaël Maucourt ◽  
Daniel Jacob ◽  
...  
Keyword(s):  
Amino Acids ◽  
Rainbow Trout ◽  
Amino Acid ◽  
Amino Acid Content ◽  
Proton Nuclear Magnetic Resonance ◽  
Free Form ◽  
Metabolomic Analysis ◽  
Nmr Metabolomics ◽  
H Nmr

Abstract The long-term effect of a plant (P)-based diet was assessed by proton nuclear magnetic resonance (1H-NMR) metabolomics in rainbow trout fed a marine fish meal (FM)–fish oil (FO) diet (M), a P-based diet and a control commercial-like diet (C) starting with the first feeding. Growth performances were not heavily altered by long-term feeding on the P-based diet. An 1H-NMR metabolomic analysis of the feed revealed significantly different soluble chemical compound profiles between the diets. A set of soluble chemical compounds was found to be specific either to the P-based diet or to the M diet. Pterin, a biomarker of plant feedstuffs, was identified both in the P-based diet and in the plasma of fish fed the P-based diet. 1H-NMR metabolomic analysis on fish plasma and liver and muscle tissues at 6 and 48 h post feeding revealed significantly different profiles between the P-based diet and the M diet, while the C diet showed intermediate results. A higher amino acid content was found in the plasma of fish fed the P-based diet compared with the M diet after 48 h, suggesting either a delayed delivery of the amino acids or a lower amino acid utilisation in the P-based diet. This was associated with an accumulation of essential amino acids and the depletion of glutamine in the muscle, together with an accumulation of choline in the liver. Combined with an anticipated absorption of methionine and lysine supplemented in free form, the present results suggest an imbalanced essential amino acid supply for protein metabolism in the muscle and for specific functions of the liver.

Evaluation of seed amino acid content and its correlation network analysis in wild soybean (Glycine soja) germplasm in Japan

2021 ◽  
Vol 19 (1) ◽  
pp. 35-43
Author(s):  
Awatsaya Chotekajorn ◽  
Takuyu Hashiguchi ◽  
Masatsugu Hashiguchi ◽  
Hidenori Tanaka ◽  
Ryo Akashi
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Free Amino Acid ◽  
Acid Content ◽  
Free Amino ◽  
Glycine Soja ◽  
Wild Soybean ◽  
Amino Acid Content ◽  
Soybean Seeds ◽  
Free Amino Acid Content

AbstractWild soybean (Glycine soja) is a valuable genetic resource for soybean improvement. Seed composition profiles provide beneficial information for the effective conservation and utilization of wild soybeans. Therefore, this study aimed to assess the variation in free amino acid abundance in the seeds of wild soybean germplasm collected in Japan. The free amino acid content in the seeds from 316 accessions of wild soybean ranged from 0.965 to 5.987 mg/g seed dry weight (DW), representing a 6.2-fold difference. Three amino acids had the highest coefficient of variation (CV): asparagine (1.15), histidine (0.95) and glutamine (0.94). Arginine (0.775 mg/g DW) was the predominant amino acid in wild soybean seeds, whereas the least abundant seed amino acid was glutamine (0.008 mg/g DW). A correlation network revealed significant positive relationships among most amino acids. Wild soybean seeds from different regions of origin had significantly different levels of several amino acids. In addition, a significant correlation between latitude and longitude of the collection sites and the total free amino acid content of seeds was observed. Our study reports diverse phenotypic data on the free amino acid content in seeds of wild soybean resources collected from throughout Japan. This information will be useful in conservation programmes for Japanese wild soybean and for the selection of accessions with favourable characteristics in future legume crop improvement efforts.

Tissue Deposition of Zinc from a Zinc Chelate and from Inorganic Zinc in Rats

1994 ◽  
Vol 1994 ◽  
pp. 171-171 ◽  
Author(s):  
Ronan Power ◽  
Kevin Cashman ◽  
Albert Flynn
Keyword(s):  
Amino Acids ◽  
Tissue Distribution ◽  
Wistar Rats ◽  
Gel Filtration ◽  
Normal Diet ◽  
Farm Animals ◽  
Trace Minerals ◽  
Inorganic Salts ◽  
Male Wistar Rats ◽  
Differential Tissue

Some reports have suggested differential tissue deposition of dietary trace minerals such as Zinc (Zn) when supplied to farm animals either chelated to amino acids or as inorganic salts. To test this hypothesis, an experiment was conducted to determine the ultimate tissue distribution of Zinc in rats fed either a radioactively-labeled 65Zn-chelate or 65ZnSO4. The 65Zn-chelate was prepared by heating a solution of 65ZnSO4 and an equimolar mixture of glycine and methionine for 5 minutes at 90°C. The resulting chelate was then separated from unincorporated 65ZnSO4 by gel filtration chromatography. Ten 25-d old male wistar rats (mean weight 34.5 g) were randomized by weight into two groups (n = 5/group), fasted for 18 hours and given 0.4 ml (8 μg Zn, 1 μCi65Zn) of one or other labelled solution by gavage. Four hours later, animals were returned to their normal diet for the duration of the experiment. The 65Zn activity of the animals was determined two hours after administration and daily thereafter for 7 days.

Characterization of an isozyme of S-adenosylmethionine synthetase from rat liver

1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Enzymatic Synthesis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Aromatic Amino Acids ◽  
Adenosylmethionine Synthetase ◽  
Cellulose Column

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.

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