Oxygen and Temperature Effect on Continuous Citric Acid Secretion in Candida Oleophila
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Microbial Enzymes and Their Applications in Industries and Medicine
BioMed Research International Microbial Enzymes and Their Applications in Industries and Medicine Guest Editors: Periasamy Anbu, Subash C. B. Gopinath, Arzu Coleri Cihan, and Bidur Prasad Chaulagain Microbial Enzymes and Their Applications in Industries and Medicine BioMed Research International Microbial Enzymes and Their Applications in Industries and Medicine Guest Editors: Periasamy Anbu, Subash C. B. Gopinath, Arzu Coleri Cihan, and Bidur Prasad Chaulagain Copyright © 2013 Hindawi Publishing Corporation. All rights reserved. This is a special issue published in “BioMed Research International.” All articles are open access articles distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Contents Microbial Enzymes and Their Applications in Industries and Medicine,PeriasamyAnbu, Subash C. B. Gopinath, Arzu Coleri Cihan, and Bidur Prasad Chaulagain Volume 2013, Article ID 204014, 2 pages Effect of C/N Ratio and Media Optimization through Response Surface Methodology on Simultaneous Productions of Intra- and Extracellular Inulinase and Invertase from Aspergillus niger ATCC 20611, Mojdeh Dinarvand, Malahat Rezaee, Malihe Masomian, Seyed Davoud Jazayeri, Mohsen Zareian, Sahar Abbasi, and Arbakariya B. Ariff Volume 2013, Article ID 508968, 13 pages A Broader View: Microbial Enzymes and Their Relevance in Industries, Medicine, and Beyond, Neelam Gurung, Sumanta Ray, Sutapa Bose, and Vivek Rai Volume 2013, Article -
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). -
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. -
Raw Materials for Industrial Fermentation Bioprocesses Improvement
Research Doctorate School in BIOmolecular Sciences Ph.D. in BIOMATERIALS - (2010-2012) RAW MATERIALS FOR INDUSTRIAL FERMENTATION BIOPROCESSES IMPROVEMENT Sergio Pirato Supervisor: Prof. Emo Chiellini Tutor: Dr. Federica Chiellini Laboratory of Bioactive Polymeric Materials for Biomedical and Environmental Applications (BIOlab) - Department of Chemistry & Industrial Chemistry, University of Pisa (Italy). 2 Sergio Pirato – PhD Thesis Sergio Pirato – PhD Thesis 3 Dietro ogni problema c'è un'opportunità. G. Galilei 4 Sergio Pirato – PhD Thesis Foreword: The present doctorate thesis in Biomaterials was formally started at the Laboratory of Bioactive Polymeric Materials for Biomedical and Environmental Application (BIOlab), at the University of Pisa, from January to September 2010. After 9 months spent at the BIOlab, the opportunity to continue the PhD as an external candidate at the Novartis Vaccines and Diagnostics came, on a topic of interest for the Biomaterials Doctorate Course. Hence, it was decided also with the consensus of the Tutor & Supervisor to continue the undertaken Doctorate Course as an external candidate without the Institutional Grant originally supplied by the University of Pisa and with the Thesis work plan accepted and endorsed by the Company itself. Therefore the thesis is consisting substantially in two parts: the first minor part is a study aiming at the development of a polymeric product suitable for the formulation of nanoparticle drug delivery systems; the second part, upon the consensus of the Supervisor and Tutor, focused on studies on raw material of biologic origin for the improvement of fermentation bioprocesses. Sergio Pirato – PhD Thesis 5 Table of Content CHAPTER 1 Aim of the Work………………………………………………………………………………. 9 1. -
Improving Citric Acid Production of an Industrial Aspergillus Niger CGMCC
Xue et al. Microb Cell Fact (2021) 20:168 https://doi.org/10.1186/s12934-021-01659-3 Microbial Cell Factories RESEARCH Open Access Improving citric acid production of an industrial Aspergillus niger CGMCC 10142: identifcation and overexpression of a high-afnity glucose transporter with diferent promoters Xianli Xue1†, Futi Bi1,2†, Boya Liu1,2, Jie Li1, Lan Zhang1, Jian Zhang1, Qiang Gao1 and Depei Wang1,2,3* Abstract Background: Glucose transporters play an important role in the fermentation of citric acid. In this study, a high- afnity glucose transporter (HGT1) was identifed and overexpressed in the industrial strain A. niger CGMCC 10142. HGT1-overexpressing strains using the PglaA and Paox1 promoters were constructed to verify the glucose transporter functions. Result: As hypothesized, the HGT1-overexpressing strains showed higher citric acid production and lower residual sugar contents. The best-performing strain A. niger 20-15 exhibited a reduction of the total sugar content and residual reducing sugars by 16.5 and 44.7%, while the fnal citric acid production was signifcantly increased to 174.1 g/L, representing a 7.3% increase compared to A. niger CGMCC 10142. Measurement of the mRNA expression levels of relevant genes at diferent time-points during the fermentation indicated that in addition to HGT1, citrate synthase and glucokinase were also expressed at higher levels in the overexpression strains. Conclusion: The results indicate that HGT1 overexpression resolved the metabolic bottleneck caused by insufcient sugar transport and thereby improved the sugar utilization rate. This study demonstrates the usefulness of the high- afnity glucose transporter HGT1 for improving the citric acid fermentation process of Aspergillus niger CGMCC 10142. -
Citric Acid Cycle
Lecture: 4 Biochemistry Anwar J Almzaiel Citric acid cycle Citric acid cycle (Krebs cycle, tricarboxylic acid cycle) is a series of reactions in mitochondria that bring about the catabolism of acetyl residues, liberating hydrogen equivalents, which upon oxidation lead to the release of most the energy of tissues fuels. The major function of cycle is to act as the final common pathway for oxidation of fatty acids, carbohydrates and proteins. It includes the electron transport system, the energy is produced in large amounts. Several of these processes are carried out in many tissues but the liver is the only tissue in which all occur to a significant extent, thus it is lethal when large numbers of hepatic cell are damaged or replaced by connective tissue, as in acute hepatitis and cirrhosis respectively. This cycle is found in the mitochondria. It starts with pyruvic acid which comes from cytoplasm to the mitochondria to be converted with coenzymes (NAD) and enzymes into acetyl COA + Pyruvic acid + COA +NAD Acetyl CoA +NADH+ CO2 The enzymes needed are found in the mitochondria close to the enzymes of the respiratory chain The oxidation of pyruvate to acetyl-CoA Before pyruvate can enter the citric acid cycle, it must be transported into the mitochondria via a special pyruvate transporter that aids its passage across the inner mitochondrial membrane. Within the mitochondria, pyruvate is oxdatively decarboxylated into acetyl COA. The conversion of pyruvic acid to acetyl COA involves 5 types of reaction and each reaction is catalysed by different enzyme system these enzymes act as a multi enzyme system (complex). -
Design and Preparation of Media for Fermentation
DESIGN AND PREPARATION OF MEDIA FOR FERMENTATION Done by: Sreelakshmi S Menon Dept of Biotechnology Fermentation Fermentation is the process in which a substance breaks down into a simpler substance using organism. Its biochemical meaning relates to the generation of energy by the catabolism of organic compounds. Fermentation is a word that has many meanings for the microbiologist: 1. Any process involving the mass culture of microorganisms, either aerobic or anaerobic. 2. Any biological process that occurs in the absence of O2. TYPES OF FERMENTATION Continuous fermentation Fed-batch fermentation Batch fermentation Submerged Liquid Fermentations Submerged liquid fermentations are traditionally used for the production of microbially derived enzymes. Submerged fermentation involves submersion of the microorganism in an aqueous solution containing all the nutrients needed for growth. Fermentation takes place in large vessels (fermenter) with volumes of up to 1,000 cubic metres. The fermentation media sterilises nutrients based on renewable raw materials like maize, sugars and soya. Most industrial enzymes are secreted by microorganisms into the fermentation medium in order to break down the carbon and nitrogen sources. Batch-fed and continuous fermentation processes are common. In the batch-fed process, sterilised nutrients are added to the fermenter during the growth of the biomass. Cont.. In the continuous process, sterilised liquid nutrients are fed into the fermenter at the same flow rate as the fermentation broth leaving the system. Parameters like temperature, pH, oxygen consumption and carbon dioxide formation are measured and controlled to optimize the fermentation process. Next in harvesting enzymes from the fermentation medium one must remove insoluble products, e.g. -
Biotechnology Past, Present and Potential
4i-"l 11- HONORARY FELLOW'S ADDRESS TO IFST 20TH ANNIVERSARY SYMPOSIUM - 16 OCT. 1985 MANCHESTER, ENGLAND IDRC - 1-lb BIOTECHNOLOGY PAST, PRESENT AND POTENTIAL BY: JOSEPH H. HULSE* BIOTECHNOLOGY: ANCIENT AND MODERN Louis Pasteur wrote "There are no applied sciences; there are only applications of science...The study of the application of science is very easy to anyone who is master of the theory". A few years later Lord Kelvin instructed us that "If you can measure that of which you speak, and can express it by a number, you know something of your subject. But if you cannot measure it, your knowledge is meagre and unsatisfactory." It would indeed be interesting to know what the spirits of these distinguished scientists are thinkinq about "Biotechnology" which takes within its broad embrace remarkable new knowledge in cell and molecular biology; some very ancient technologies; together with a large swatch of enpirical observations and discoveries, many of which remain far distant from viable technological application. Fermentation technologies have a very long history: beer, wine, bread and cheese having been around as long as cereals and vine fruits have been harvested and animais have been milked. Homer described wine as a qift from the Gods and Ecclesiasticus wisely advised that "From the beginning wine was created to make men joyful, not to make them drunk." Though ethanolic fermentations have been most pervasive, lactic and other acidic fermentations have appeared in greater diversity, particularly in traditional domestic processes of preservation. The ancient Sumerians 7,000 years ago converted all their milk into cheese in the stated belief that had God intended mankind to have clean milk to drink he would have placed the udders at the front end of the cow.