Citric Acid Cycle
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The Mechanism of Aconitase Action Iii. Kinetic Analysis Using Dl-Isocitric Acid-2-C 14
TheJournal of Biochemistry, Vol.41, No. 5, 1954 THE MECHANISM OF ACONITASE ACTION III. KINETICANALYSIS USING DL-ISOCITRIC ACID-2-C14 By JUN-ICHI TOMIZAWA (Fromthe NationalInstitute of Health,Tokyo) (Receivedfor publication,June 24, 1954). In the previous reports (1, 2, 3), the author came to a conclusion that aconitase would be a single enzyme, and one enzyme and one activated complex theory was proposed. In the present communication, some additional proofs of the theory using labeled substrates will be reported. EXPERIMENTAL The Enzyme Preparation and the Methods of Analysis The preparation of the rabbit liver enzyme was the same as reported previously (1) except it was centrifuged at 120,000 x g for 30 minutes and the supernatant was kept. General properties of the enzyme preparation were not changed by this treat ment. The preparation and the analysis of non-labeled substrates were previously reported. The Preparation of DL-Isocitric Acid-2-C14 Usually DL-isocitric acid was prepared by hydrolysis of trichloromethylparaconic acid. It was, however, rather difficult to prepare the carbon-2 labeled compound by this method. Therefore, the compound was prepared by an entirely new process. Ethyl Formate-C14•\Formic acid was prepared by the usual method from labeled barium carbonate. Its ethyl ester was prepared by esterification of the sodium salt as was done in the preparation of its methyl ester according to Me1viIIeetal. (4, 5). The yield was about 80per cent. Diethyl Formyl-succinate-l-C14-This compound was prepared by the method of Sugazawa (6). 0.6g. of ethyl formate-C14 and 1.2g. -
EFFECT of INOCULUM on KINETICS and YIELD of CITRIC ACIDS PRODUCTION on GLUCOSE by Yarrowia Lipolytica A-101
.\ C T A ALIMENTARIA POLONI C A Vol. XVII/ XLI ! No. 2 1991 MARIA WOJTATOWICZ WALDEMAR RYMOWICZ EFFECT OF INOCULUM ON KINETICS AND YIELD OF CITRIC ACIDS PRODUCTION ON GLUCOSE BY Yarrowia Lipolytica A-101 Departmcnt of Biotechnology and Food Microbiology, Academy of Agriculture, Wrocław Key word s: Yarrowia fipo(1 'fin,1 /\-1 O1 , citric and isodtric acid, glucosc, nitrogen deficient medium The cffcct of two differcnt inocula on growth and production parameters in citric acid fcrmcntation (on glucose) by Yarrowia lipolytica A-101 was studied. For inoculum prcpared in full growth medium the total acids yield was 5-12% higher and · biomass yield about 10 % higher than for inoculum prepared in a nitrogen-deficient medium. The latter inoeulum, however, !cd to about 10-30% highcr acid producti.on and glucosc consumption rates. Until the early I 970s practically the only organisms used to produce citric acid were Aspergillus niger and a few other fungi. Today we know that many kinds of yeasts can accumulate substantial amounts of citric acid in their growth media. The most cfficient citric acid producers belong to the Candida genus, and the strains used most often are C. lipolytica, C. zeylanoides, C. parapsilosis, C. tropicalis, C. guilliermondii, C. oleophila, C. petrophilum and C. intermedia [10]. Yeasts can produce citric acid more rapidly than fungi and in a greater variety of substrates including n-alkanes, n-alkenes, glucose, molasses, acetate, alcohols, fatty acids and natura) oils [4, 8, 9, 17, 18]. The product yield on n-alkanes and vegetable oils may be as high as 1.6 g/g [4, 9]; on glucose it is usually comparable to that in proccsses involving filamentous molds [4-6]. -
D-Isocitric Acid (D-Isocitrate)
www.megazyme.com D-ISOCITRIC ACID (D-ISOCITRATE) ASSAY PROCEDURE K-ISOC 11/19 (*100 Manual Assays per Kit) or (1000 Auto-Analyser Assays per Kit) or (1000 Microplate Assays per Kit) * The number of tests per kit can be doubled if all volumes are halved © Megazyme 2019 INTRODUCTION: D-Isocitric acid is an organic acid found in most fruit juices. It is an important marker in multicomponent procedures for the evaluation of authenticity and quality of fruit products; high citric/isocitric acid ratios can be used as an indicator of citric acid addition in some juices. PRINCIPLE: D-Isocitric acid is oxidised by nicotinamide-adenine dinucleotide + phosphate (NADP ) to 2-oxoglutarate and CO2 in the presence of isocitrate dehydrogenase (ICDH), with the formation of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) (1). (ICDH) + + (1) D-Isocitric acid + NADP 2-oxoglutarate + CO2 + NADPH + H The amount of NADPH formed in this reaction is stoichiometric with the amount of D-isocitric acid. It is the NADPH which is measured by the increase in absorbance at 340 nm. Bound D-isocitric acid is released by alkaline hydrolysis (2), (3), and then measured using the same principle (1). (pH 9-10) (2) D-Isocitric acid ester + H2O D-isocitric acid + alcohol (pH 9-10) (3) D-Isocitric acid lactone + H2O D-isocitric acid SPECIFICITY, SENSITIVITY, LINEARITY AND PRECISION: The assay is specific for D-isocitric acid. D-malic acid, L-lactic acid, L-aspartic acid and fumaric acid do not react. The smallest differentiating absorbance for the assay is 0.005 absorbance units. This corresponds to 0.177 mg/L of sample solution at the maximum sample volume of 2.00 mL (or to 3.54 mg/L with a sample volume of 0.1 mL). -
Class 11 Biology Chapter- 13 Respiration in Plants
CLASS 11 BIOLOGY CHAPTER- 13 RESPIRATION IN PLANTS CELLULAR RESPIRATION: The process of conversion of the chemical energy of organic substances into a metabolically usable energy within living cells is called cellular respiration. TYPES OF CELLULAR RESPIRATION: (i) Aerobic respiration: The process of respiration which requires molecular oxygen. (ii) Anaerobic respiration: The process of respiration which does not require molecular oxygen and occurs in the cytoplasm. MECHANISM OF RESPIRATION: Following are the steps- 1. GLYCOLYSIS / EMP Pathway: It involves a series of closely integrated reactions in which hexose sugars(usually glucose) are converted into pyruvic acid. It is common in both aerobic and anaerobic reactions. It occurs in the cytoplasm. It does not require oxygen. Gollowing are the steps of GLYCLOLYSIS: (i) Conversion of glucose to Fructose-1,6-diphosphate: First phosphorylation: Glucose is converted to Glucose -6-phosphate in the presence of enzyme hexokinase and Mg++ ions and energy in the form of ATP. Isomerization: Glucose-6-phosphate is converted to Fructose-6-phosphate in the presence of phosphohexoisomerase. Second phosphorylation: Fructose-6-phosphate is converted to Fructose-1,6- diphosphate by the use of energy in the form of ATP. (ii) Formation of pyruvic acid from fructose -1,6-diphosphate: Cleavage: Fructose-1,6-diphosphate splits into 3-phosphoglyceraldehyde and Dihydroxyacetone phosphate in the presence of enzyme aldolase. Phosphorylation and oxidative dehydrogenase: 3-phosphoglyceraldehyde is converted to 1,3-biphosphoglyceric acid. ATP generation(first): 1,3-biphosphoglyceric acid is converted to 3-phosphoglyceric acid in the presence of Mg++ and phosphoglycerokinase . Isomerization: 3-phosphoglyceric acid is converted to 2-phosphoglyceric acid in the presence of Mg++. -
Detection and Formation Scenario of Citric Acid, Pyruvic Acid, and Other Possible Metabolism Precursors in Carbonaceous Meteorites
Detection and formation scenario of citric acid, pyruvic acid, and other possible metabolism precursors in carbonaceous meteorites George Coopera,1, Chris Reeda, Dang Nguyena, Malika Cartera, and Yi Wangb aExobiology Branch, Space Science Division, National Aeronautics and Space Administration-Ames Research Center, Moffett Field, CA 94035; and bDevelopment, Planning, Research, and Analysis/ZymaX Forensics Isotope, 600 South Andreasen Drive, Suite B, Escondido, CA 92029 Edited by David Deamer, University of California, Santa Cruz, CA, and accepted by the Editorial Board July 1, 2011 (received for review April 12, 2011) Carbonaceous meteorites deliver a variety of organic compounds chained three-carbon (3C) pyruvic acid through the eight-carbon to Earth that may have played a role in the origin and/or evolution (8C) 7-oxooctanoic acid and the branched 6C acid, 3-methyl- of biochemical pathways. Some apparently ancient and critical 4-oxopentanoic acid (β-methyl levulinic acid), Fig. 1, Table S1. metabolic processes require several compounds, some of which 2-methyl-4-oxopenanoic acid (α-methyl levulinic acid) is tenta- are relatively labile such as keto acids. Therefore, a prebiotic setting tively identified (i.e., identified by mass spectral interpretation for any such individual process would have required either a only). As a group, these keto acids are relatively unusual in that continuous distant source for the entire suite of intact precursor the ketone carbon is located in a terminal-acetyl group rather molecules and/or an energetic and compact local synthesis, parti- than at the second carbon as in most of the more biologically cularly of the more fragile members. -
Challenges in Analysis of Hydrophilic Metabolites Using Chromatography Coupled with Mass Spectrometry
Journal of Analysis and Testing https://doi.org/10.1007/s41664-020-00126-z REVIEW Challenges in Analysis of Hydrophilic Metabolites Using Chromatography Coupled with Mass Spectrometry Qingyu Hu1,2,3 · Huiru Tang3 · Yulan Wang4 Received: 2 February 2020 / Accepted: 26 March 2020 © The Nonferrous Metals Society of China 2020 Abstract Hydrophilic metabolites play important roles in cellular energy metabolism, signal transduction, immunity. However, there are challenges in both identifcation and quantifcation of the hydrophilic metabolites due to their weak interactions with C18-reversed-phase liquid chromatography (RPLC), leading to poor retention of hydrophilic metabolites on the columns. Many strategies have been put forward to increase the retention behavior of hydrophilic metabolites in the RPLC system. Non- derivatization methods are mainly focused on the development of new chromatographic techniques with diferent separation mechanisms, such as capillary electrophoresis, ion-pairing RPLC etc. Derivatization methods improve the hydrophobicity of metabolites and can enhance the MS response. This review mainly focused on the illustration of challenges of LCMS in the analysis of hydrophilic metabolomics feld, and summarized the non-derivatization and derivatization strategies, with the intention of providing multiple choices for analysis of hydrophilic metabolites. Keywords Hydrophilic metabolites · Hydrophilic interaction chromatography · Ion-pairing reversed-phase liquid chromatography · Ion chromatography · Capillary electrophoresis · -
Continuous Citric Acid Secretion by a High Specific Ph Dependent Active Transport System in Yeast Candida Oleophila ATCC 20177
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.8 No.2, Issue of August 15, 2005 © 2005 by Pontificia Universidad Católica de Valparaíso -- Chile Received November 22, 2004 / Accepted March 28, 2005 RESEARCH ARTICLE Continuous citric acid secretion by a high specific pH dependent active transport system in yeast Candida oleophila ATCC 20177 Savas Anastassiadis*# Department of Environmental Engineering School of Engineering Democritus University of Thrace 67100 Xanthi, Greece E-mail: [email protected] Hans-Jürgen Rehm Institute of Microbiology University of Münster Corrensstr. 3, 48149 Münster, Germany (retired Professor) Website: http://www.greekbiotechnologycenter.gr Financial support: Part of the work that has been carried out at the Institute of Biotechnology 2 of Research Centre Jülich (Germany) was financed by Haarmann and Reimer, a daughter company of the company Bayer, Leverkusen, Germany. Keywords: active citrate export, citric acid fermentation, energy consuming citric acid secretion, specific active transport system. Present address: #Research in Biotechnology, Co., Vat. #: 108851559. Avgi/Sohos, 57002 Thessaloniki, Greece; Tel. +30-2395-051324; +30-6973- 801395 (cellular); Tel./Fax. +30-2395-051470, E-mail: [email protected]. The pH influence on continuous citric acid secretion was similarities between A. niger and yeast strains in investigated in Candida oleophila ATCC 20177 (var.) mechanism of citric acid synthesis, however, differences + under NH4 limiting state steady conditions, using still exist in terms of triggering out and regulation of citrate glucose. Highest citric acid concentration of 57.8 g/l, overproduction. Many models have been developed citrate/isocitrate ratio of 15.6, space-time yield of 0.96 describing the biochemistry of citrate synthesis, using g/(l x hr) and biomass specific productivity of 0.041 g/(g glucose and other carbon sources, however a complete x hr) were obtained at pH 5 and 60 hrs residence time. -
Antibiotics As a Feed Additive Promote Growth of Chickens by Modulating the Gut Microbiome
Antibiotics as a Feed Additive Promote Growth of Chickens by Modulating the Gut Microbiome Xiangkai Li ( [email protected] ) Lanzhou University https://orcid.org/0000-0002-5704-9791 Ying Wu Lanzhou University Liang Peng Lanzhou University Rong Han Lanzhou University pengya Feng Lanzhou University Aman Khan Lanzhou University Pu Liu Lanzhou University Research Keywords: Antibiotic, Gut microbiota, Metabonomic, Lipid and amid acid metabolism, Immune response Posted Date: June 7th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-571302/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/31 Abstract Background: Antibiotics widely used as growth promoters in the agricultural industry, but their mechanisms have not been fully explored. Antibiotics has a connection effect on the gut microbiome which plays a vital role in host metabolism and immune response. Here, we investigated the association of antibiotic and gut microbiome in broiler chicken. Methods: Polymyxin (PMX) and amoxicillin (AMX) were selected as feed additives in broiler chicken, and gut bacterial composition assessed by quantitative real-time PCR and high-throughput sequencing of bacterial 16S rRNA. Metabolome and lipidomic of feces and serum samples were analyzed using liquid chromatograpy-tandem mass spectrometry (LC-MS). We further assessed changes in the microbiota and metabolism which underwent antibiotics treatment. Results: The administration of antibiotic increasing, the average weight of chickens by up to 4.9% and altered number and structure of the intestinal microora compared to the un-treated group. The bacterial component of gut microbiota in antibiotic groups was showed a lower prevalence of Firmicutes and Bacteroidetes phyla and higher prevalence of a high diversity (Proteobacteria). -
FOOD ACIDS in the HUMAN BODY and Krebs Cycle
FOOD ACIDS IN THE HUMAN through the intestinal wall into the BODY bloodstream in the form of amino acids, mono-saccharides, glycerol and emulsified The acidulants, malic acid, fumaric acid, and fatty acids. The residue passes through the citric acid, occur naturally in all living large intestine, where the main activities of organisms of both the vegetable and animal water absorption and proliferation of bacteria kingdom. They all play an integral part in the takes place, and is in due course excreted. normal respiratory processes in living cells. These compounds are formed in the A compound absorbed into the blood stream, metabolic cycles in the cells of plants and has one of three fates thereafter. animals. The acids provide the cell with energy and the carbon skeletons for the 1. It may interfere with the normal workings formation of amino acids. This takes place in of the body and will - unless it kills the the Krebs cycle. host first - be converted, probably in the liver, to some innocuous compound which Food consumption by man is essential for will be filtered out by the kidneys and providing energy to keep the engine of the excreted in the urine. human body running. A bite of food first gets broken up physically in the mouth and mixes 2. It may simply not fit any metabolic pattern with the alkaline saliva, which initiates in the body and, when it reaches the hydrolysis of starch and sugar. It is then kidneys, will be filtered out and excreted swallowed and passes into the stomach in the urine. -
Interpretive Guide
INTERPRETIVE GUIDE Contents INTRODUCTION .........................................................................1 NUTREVAL BIOMARKERS ...........................................................5 Metabolic Analysis Markers ....................................................5 Malabsorption and Dysbiosis Markers .....................................5 Cellular Energy & Mitochondrial Metabolites ..........................6 Neurotransmitter Metabolites ...............................................8 Vitamin Markers ....................................................................9 Toxin & Detoxification Markers ..............................................9 Amino Acids ..........................................................................10 Essential and Metabolic Fatty Acids .........................................13 Cardiovascular Risk ................................................................15 Oxidative Stress Markers ........................................................16 Elemental Markers ................................................................17 Toxic Elements .......................................................................18 INTERPRETATION-AT-A-GLANCE .................................................19 REFERENCES .............................................................................23 INTRODUCTION A shortage of any nutrient can lead to biochemical NutrEval profile evaluates several important biochemical disturbances that affect healthy cellular and tissue pathways to help determine nutrient -
Carboxylic Acids
13 Carboxylic Acids The active ingredients in these two nonprescription pain relievers are derivatives of arylpropanoic acids. See Chemical Connections 13A, “From Willow Bark to Aspirin and Beyond.” Inset: A model of (S)-ibuprofen. (Charles D. Winters) KEY QUESTIONS 13.1 What Are Carboxylic Acids? HOW TO 13.2 How Are Carboxylic Acids Named? 13.1 How to Predict the Product of a Fischer 13.3 What Are the Physical Properties of Esterification Carboxylic Acids? 13.2 How to Predict the Product of a B-Decarboxylation 13.4 What Are the Acid–Base Properties of Reaction Carboxylic Acids? 13.5 How Are Carboxyl Groups Reduced? CHEMICAL CONNECTIONS 13.6 What Is Fischer Esterification? 13A From Willow Bark to Aspirin and Beyond 13.7 What Are Acid Chlorides? 13B Esters as Flavoring Agents 13.8 What Is Decarboxylation? 13C Ketone Bodies and Diabetes CARBOXYLIC ACIDS ARE another class of organic compounds containing the carbonyl group. Their occurrence in nature is widespread, and they are important components of foodstuffs such as vinegar, butter, and vegetable oils. The most important chemical property of carboxylic acids is their acidity. Furthermore, carboxylic acids form numerous important derivatives, including es- ters, amides, anhydrides, and acid halides. In this chapter, we study carboxylic acids themselves; in Chapters 14 and 15, we study their derivatives. 457 458 CHAPTER 13 Carboxylic Acids 13.1 What Are Carboxylic Acids? Carboxyl group A J COOH The functional group of a carboxylic acid is a carboxyl group, so named because it is made group. up of a carbonyl group and a hydroxyl group (Section 1.7D). -
Metabolomic Analysis of Trichophyton Rubrum and Microsporum Canis
www.nature.com/scientificreports OPEN Metabolomic analysis of Trichophyton rubrum and Microsporum canis during keratin degradation Anita Ciesielska 1*, Anna Kawa1, Katarzyna Kanarek1, Adrian Soboń 1 & Rafał Szewczyk 2 Keratin is important and needed for the growth of dermatophytes in the host tissue. In turn, the ability to invade keratinised tissues is defned as a pivotal virulence attribute of this group of medically important fungi. The host–dermatophyte interaction is accompanied by an adaptation of fungal metabolism that allows them to adhere to the host tissue as well as utilize the available nutrients necessary for their survival and growth. Dermatophyte infections pose a signifcant epidemiological and clinical problem. Trichophyton rubrum is the most common anthropophilic dermatophyte worldwide and its typical infection areas include skin of hands or feet and nail plate. In turn, Microsporum canis is a zoophilic pathogen, and mostly well known for ringworm in pets, it is also known to infect humans. The aim of the study was to compare the intracellular metabolite content in the T. rubrum and M. canis during keratin degradation using liquid chromatography system coupled with tandem mass spectrometer (LC-MS/MS). The metabolite “fngerprints” revealed compounds associated with amino acids metabolism, carbohydrate metabolism related to the glycolysis and the tricarboxylic acid cycle (TCA), as well as nucleotide and energy metabolism. The metabolites such as kynurenic acid, l-alanine and cysteine in case of T. rubrum as well as cysteine and ribofavin in case of M. canis were detected only during keratin degradation what may suggest that these compounds may play a key role in the interactions of T.