DOCSLIB.ORG

I. Demonstration of Pentose Isomerase, Pentulokinase, and Pentitol Dehydrogenase Enzyme Families' R

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
I. Demonstration of Pentose Isomerase, Pentulokinase, and Pentitol Dehydrogenase Enzyme Families' R Jouwra OF BACTERIOLOGY Vol. 88, No. 4, p. 838-844 October, 1964 Copyright § 1964 American Society for Microbiology Printed in U.S.A. METABOLISM OF PENTOSES AND PENTITOLS BY AEROBACTER AEROGENES I. DEMONSTRATION OF PENTOSE ISOMERASE, PENTULOKINASE, AND PENTITOL DEHYDROGENASE ENZYME FAMILIES' R. P. MORTLOCK2 AND W. A. WOOD Department of Biochemistry, Michigan State University, East Lansing, Michigan Received for publication 1 April 1964 ABSTRACT among bacteria in that it can grow on all eight MORTLOCK, R. P. (Michigan State University, of the aldopentoses (Simpson, unpublished data) East Lansing) AND W. A. WOOD. Metabolism of and all four of the pentitols. This versatility is pentoses and pentitols by Aerobacter aerogenes. I. more striking because many of these compounds Demonstration of pentose isomerase, pentuloki- rarely, if ever, occur in nature. Fermentation nase, and pentitol dehydrogenase enzyme families. studies with specifically labeled pentoses showed J. Bacteriol. 88:838-844. 1964.-Aerobacter aero- that several of the pentoses yield the same fer- genes PRL-R3 is capable of utilizing as sole sub- mentation products and with identical labeling strates for energy and growth seven of the eight patterns (Neish and Simpson, 1954; Altermatt, aldopentoses and all of the four pentitols. Growth Simpson, and Neish, 1955). Thus, the same gen- upon media containing either D-xylOse, L-arabi- nose, D-ribose, D-arabitol, or ribitol occurred eral metabolic routes must be involved for the within 24 hr at 26 C. When D-arabinose or L-arabi- fermentation of these compounds. The strategy tol were used as growth substrates, growth was for pentose utilization as elucidated in many complete within 2 days; 4 days were required for organisms involves isomerization of an aldopen- growth on D-lyxose or xylitol, and 3 to 4 weeks for tose to (one of the four) ketopentoses, phos- growth upon L-xylose. The versatility of this phorylation of the ketopentoses, and epimeriza- strain of A. aerogenes is due to an ability to syn- tion of ketopentose-5-phosphates to D-xylu- thesize in the presence of appropriate carbohy- lose-5-phosphate. Pentitols are utilized by drates (inducers) families of enzymes which cata- dehydrogenation to the ketopentose, but the lyze the metabolism of the carbohydrates (i.e., remaining steps are identical. families of pentitol dehydrogenases, aldopentose isomerases, and pentulokinases). The specificity of For A. aerogenes, these processes accommodate induction for members of the enzyme families was the whole range of C5 structures. Current infor- found to vary, and cross induction of enzyme ac- mation on these pathways is summarized in Fig. tivity was common, especially among the pentitol 1. The individual reactions in the utilization of dehydrogenases. Ribitol dehydrogenase activity L-arabinose, L-xylose, L-lyxose, and the pentitols was detected in extracts of cells grown on all of were intensively studied in this strain. As shown the above carbohydrates with the exception of by Simpson, Wolin, and Wood [(1958); see also D-xylose, L-arabinose, and D-ribose. The ribitol Burma and Horecker (1958) for similar processes dehydrogenase activity of xylitol-grown cell ex- in Lactobacillus plantarum], L-arabinose is isom- tracts was fivefold higher than the activity in ex- erized to L-ribulose which is then phosphorylated tracts of ribitol-grown cells. to 4-ribulose-5-phosphate. This phosphate ester is then epimerized at carbon four to form D-xylu- Aerobacter aerogenes, PRL-R3, is unusual lose-5-phosphate, the substrate of transketolase. Similarly, L-xylose and L-lyxose are isomerized to I Contribution no. 3272 of the Michigan Agri- L-xylulose, which is then phosphorylated to L- cultural Experiment Station. A preliminary report of this investigation was presented at the Annual xylulose-5-phosphate. By epimerizations at car- Meeting of the American Society for Microbiology bons three and four, this ester is also converted at Kansas City, Mo., May, 1962. to D-xylulose-5-phosphate (Anderson and Wood, 2 Postdoctoral Fellow of the U.S. Public Health 1962a). Thus, all of these isomers are converted Service. Present address: Department of Micro- to D-xylulose-5-phosphate by epimerases. biology, University of Massachusetts, Amherst. The oxidation of ribitol to D-ribulose and of 838 VOL'. 88, 1964 ENZYME FAMILIES IN A. AEROGENES !839 D-arabitol to D-xylulose by specific nicotinamide (c) pyruvate + reduced nicotinamide. .,{f, adenine dinucleotide (NAD)-linked dehydro- adenine dinucleotide (NADIb) genases has been documented (Fromm, 1958; Wood, McDonough, and Jacobs, 1961; Lin, 1961), lactic dehydrogenae >lactate + NAD as have the oxidations of L-arabitol to L-xylulose and xylitol to D-xylulose (Fossitt et al., 1964). For determination of pentitol dehydrogenase For D-ribose, a difference in detail exists, in that activity, the same reaction mixture was used, phosphorylation precedes isomerization. except that ATP was omitted and the rate of Growth of A. aerogenes on all of these five- pentulose reduction (NADH oxidation) at 340 m,u carbon structures implies an unusual ability to was determined. To measure kinase activity in synthesize several families of enzymes, with the the presence of large amounts of pentitol dehy- members of each famnily catalyzing the same re- drogenase activity, reduced nicotinamide adenine action, but each having a different specificity. dtnucleotide phosphate (NADPH) was substi- Because of the value of these families in studying tuted for NADH. NADPH serves as an efficient determinants of specificity, a detailed analysis of reductant for pyruvate in the presence of excess this phenomenon has been undertaken. lactic dehydrogenase, but is not utilized by NADH oxidase or the NADH-specific pentitol MATERIALS AND METHODS dehydrogenases, thereby greatly decreasing the Bacteriological. A. aerogenes, PRL-R3, was blank rate. A unit of kinase or pentitol dehy- grown aerobically at 25 C on a minimal medium drogenase activity was defined as the amount of (Anderson and Wood, 1962a) supplemented with enzyme giving an absorbancy change of 1.0 per 0.5% carbohydrate. The carbohydrate and min at 340 m,u in a reaction volume of 0.15 ml MgSO4 were autoclaved separately and added and with a light path of 1 cm. after cooling. For some experiments, the carbo- Isomerase activity was measured from the rate hydrate was sterilized by filtration through a of pentulose formation by the method of Ander- type HA Millipore filter. The viable cell determi- son and Wood (1962a); 1 unit of isomerase in 2.0 nations were obtained by plating on the above ml catalyzed the formation of 1 ,umole of pentu- medium supplemented with 1.5% Nobel Agar lose per hr at 37 C. (Difco). Total cell counts were made with a Oxygen utilization was determined by stand- Petroff-Hausser counting chamber. Turbidity ard manometric methods. Protein was estimated determinations were made in a Bausch & Lomb by the method of Lowry et al. (1951). Aldopen- Spectronic-20 colorimeter, and were converted to toses were determined by the orcinol test (Mej- dry weight of cell material with a standard curve. baum, 1939), with a 40-min heating time. Keto- For standardized growth experiments, tubes pentose was measured by the cysteine-carbazole test of Dische and Borenfreund (1951). containing 5.5 ml of medium were inoculated to Chemical. Ribitol, D-arabitol, give 2 X 106 cells per ml, inclined to 450, and xylitol, D-xylose, incubated at 26 C on a reciprocating shaker. The and D-ribose were purchased from Nutritional inoculum was washed twice with sterile salts solution prior to use. Cultures were considered D-RIBOSE kinose O-RIBOSE-5-PHOSPHATE grown when the turbidity at 620 m, reached an D-ARABINOSE isomerose isomerose kinose optical density of 1.0. D-RIBULOSE D-RIBULOSE-5-PHOSPHATE Analytical. Kinase activity was estimated by RIBITOL dehydrogenose L use of the following coupled reaction sequence 0-XYLOSE isornerose 3pimerase D-LYXOSE isomerose k kinose described by Anderson and Wood (1962b): o-XYLULOSE 11-D-XYLULOSE-5-PH SPHAT D-ARABITOL 4nit2roense (a) pentulose + adenosine triphosphate (ATP) XYLITOL dehdromse4m isoerneose kinose { 4-pinerose kinase > pentulose-5-phosphate L-ARABINOSE --- L-RIBULOSE ko L-RIBULOSE-5-PHOSPHATE L-XYLOSE isomerose + adenosine diphosphate (ADP) L-LYXOSE isoerose 3-epimerse L-XYLULOSE --- L-XYLULOSE-5-PHOSPHATE A (b) ADP + phosphoenolpyruvate L-ARABI TOL !2^dr9e~nqse FIG. Aldopentose andk/inose3-hnrs pyruvate kinase> 1. Aldopentose pentitol metabolism by pyruvate + ATP Aerobacter aerogenes. 840 MORTLOCK AND WOOD J. BACTERIOL. Biochemicals Corp., Cleveland, Ohio. r-Arab- RESULTS inose and D-arabinose were obtained from Growth characteistics. A. aerogenes, PRL-R3, Pfanstiehl Laboratories, Inc., Waukegan, Ill.; utilized as a sole source of carbon and energy for L-arabitol was purchased from Mann Research growth seven of the eight aldopentoses and all of Laboratory, New York, N.Y.; D-lyxose was pur- the pentitols. The times required to reach a pre- chased from General Biochemicals, Chagrin scribed growth level on these substrates are Falls, Ohio; L-xylose, L-lyxose, and t-xylulose shown in Table 1. With the use of a standard were prepared as described by Anderson and inoculum of D-glucose-grown cells, growth on Wood (1962a); and L-xylose was also purchased D-glucose, D-xylose, D-ribose, L-arabinose, ribitol, from General Biochemicals. D-Ribulose was pre- and D-arabitol was complete within 1 day. A pared from the o-nitrophenylhydrazone deriva- longer incubation time was required for the other tive as described by Muller, Montigel, and Reich- substrates as follows: D-arabinose and L-arabitol, stein (1937). Recrystallization of aldopentoses 2 days; D-lyxose and xylitol, 4 days; and L-xylose, and pentitols was carried out in absolute ethanol. from 3 to 4 weeks. The longer growth period on Enzymatic. To prepare cell-free extracts for these substrates was due to a lag of approxi- enzyme assays, the cells were harvested by cen- mately 15 hr with D-arabinose and L-arabitol, trifugation, washed once in 0.3 volume of water, and 3 days with D-lyxose or xylitol.
Load more

Recommended publications
  • Reactions of Saccharides Catalyzed by Molybdate Ions. XXII.* Oxidative Degradation of D-Galactose Phenylhydrazones
    Reactions of saccharides catalyzed by molybdate ions. XXII.* Oxidative degradation of D-galactose phenylhydrazones L. PETRUŠ, V. BILIK, K. LINEK, and M. MISIKOVA Institute of Chemistry, Slovak Academy of Sciences, 809 33 Bratislava Received 4 March 1977 D-Galactose phenylhydrazone was degraded with hydrogen peroxide in the presence of molybdate ions to D-lyxose in 50% yield. The oxidative degrada­ tion of D-galactose 2,5-dichlorophenylhydrazone and D-galactose 2,4-dinitro- phenylhydrazone gave D-lyxose and D-galactose in the ratio 4:1 and 1:9, respectively. 2,6-Anhydro-l-deoxy-l-nitro-D-galactitol was prepared from D-lyxose. Фенилгидразон D-галактозы разрушается перекисью водорода в при­ сутствии молибдатных ионов с 50%-ным превращением в D-ликсозу. В случае окислительной деградации 2,5-дихлорфенилгидразона D-галак­ тозы образуется D-ликсоза и D-галактоза в отношении 4:1, в случае же 2,4-динитрофенилгидразона D-галактозы в отношении 1:9. Из D-ликсозы был приготовлен 2,6-ангидро-1-дезокси-1-нитро-о-галактитол. Treatment of 1-deoxy-l-nitroalditols in alkaline medium with hydrogen peroxi­ de in the presence of molybdate ions leads to the formation of corresponding aldoses [1]. This reaction is, particularly with nitroalditols prepared from L-ribose [2] and D-glucose [3], accompanied by a parallel elimination reaction leading back to the starting aldoses. Schulz and Somogyi [4] treating L-rhamnose phenylhydra­ zone with oxygen in acetone or 2,3,4,5,6-penta-O-acetyl-D-galactose phenylhy­ drazone in benzene, obtained the corresponding 1-hydroperoxo derivatives which decomposed in alcohol solution of sodium methanolate to the corresponding pentoses.
    [Show full text]
  • An Atpase Domain Common to Prokaryotic Cell Cycle Proteins
    Proc. Natl. Acad. Sci. USA Vol. 89, pp. 7290-7294, August 1992 Biochemistry An ATPase domain common to prokaryotic cell cycle proteins, sugar kinases, actin, and hsp7O heat shock proteins (structural comparison/property pattern/remote homology) PEER BORK, CHRIS SANDER, AND ALFONSO VALENCIA European Molecular Biology Laboratory, D-6900 Heidelberg, Federal Republic of Germany Communicated by Russell F. Doolittle, March 6, 1992 ABSTRACT The functionally diverse actin, hexokinase, and hsp7O protein families have in common an ATPase domain of known three-dimensional structure. Optimal superposition ofthe three structures and alignment ofmany sequences in each of the three families has revealed a set of common conserved residues, distributed in five sequence motifs, which are in- volved in ATP binding and in a putative interdomain hinge. From the multiple sequence aliment in these motifs a pattern of amino acid properties required at each position is defined. The discriminatory power of the pattern is in part due to the use of several known three-dimensional structures and many sequences and in part to the "property" method ofgeneralizing from observed amino acid frequencies to amino acid fitness at each sequence position. A sequence data base search with the pattern significantly matches sugar kinases, such as fuco-, glucono-, xylulo-, ribulo-, and glycerokinase, as well as the prokaryotic cell cycle proteins MreB, FtsA, and StbA. These are predicted to have subdomains with the same tertiary structure as the ATPase subdomains Ia and Ha of hexokinase, actin, and Hsc7O, a very similar ATP binding pocket, and the capacity for interdomain hinge motion accompanying func- tional state changes.
    [Show full text]
  • Non-Homologous Isofunctional Enzymes: a Systematic Analysis Of
    Omelchenko et al. Biology Direct 2010, 5:31 http://www.biology-direct.com/content/5/1/31 RESEARCH Open Access Non-homologousResearch isofunctional enzymes: A systematic analysis of alternative solutions in enzyme evolution Marina V Omelchenko, Michael Y Galperin*, Yuri I Wolf and Eugene V Koonin Abstract Background: Evolutionarily unrelated proteins that catalyze the same biochemical reactions are often referred to as analogous - as opposed to homologous - enzymes. The existence of numerous alternative, non-homologous enzyme isoforms presents an interesting evolutionary problem; it also complicates genome-based reconstruction of the metabolic pathways in a variety of organisms. In 1998, a systematic search for analogous enzymes resulted in the identification of 105 Enzyme Commission (EC) numbers that included two or more proteins without detectable sequence similarity to each other, including 34 EC nodes where proteins were known (or predicted) to have distinct structural folds, indicating independent evolutionary origins. In the past 12 years, many putative non-homologous isofunctional enzymes were identified in newly sequenced genomes. In addition, efforts in structural genomics resulted in a vastly improved structural coverage of proteomes, providing for definitive assessment of (non)homologous relationships between proteins. Results: We report the results of a comprehensive search for non-homologous isofunctional enzymes (NISE) that yielded 185 EC nodes with two or more experimentally characterized - or predicted - structurally unrelated proteins. Of these NISE sets, only 74 were from the original 1998 list. Structural assignments of the NISE show over-representation of proteins with the TIM barrel fold and the nucleotide-binding Rossmann fold. From the functional perspective, the set of NISE is enriched in hydrolases, particularly carbohydrate hydrolases, and in enzymes involved in defense against oxidative stress.
    [Show full text]
  • Electronic Supplementary Information
    Electronic Supplementary Material (ESI) for Chemical Science. This journal is © The Royal Society of Chemistry 2019 Electronic Supplementary Information Poly(ionic liquid)s as a Distinct Receptor Material to Create Highly- Integrated Sensing Platform for Efficiently Identifying a Myriad of Saccharides Wanlin Zhang, Yao Li, Yun Liang, Ning Gao, Chengcheng Liu, Shiqiang Wang, Xianpeng Yin, and Guangtao Li* *Corresponding authors: Guangtao Li ([email protected]) S1 Contents 1. Experimental Section (Page S4-S6) Materials and Characterization (Page S4) Experimental Details (Page S4-S6) 2. Figures and Tables (Page S7-S40) Fig. S1 SEM image of silica colloidal crystal spheres and PIL inverse opal spheres. (Page S7) Fig. S2 Adsorption isotherm of PIL inverse opal. (Page S7) Fig. S3 Dynamic mechanical analysis and thermal gravimetric analysis of PIL materials. (Page S7) Fig. S4 Chemical structures of 23 saccharides. (Page S8) Fig. S5 The counteranion exchange of PIL photonic spheres from Br- to DCA. (Page S9) Fig. S6 Reflection and emission spectra of spheres for saccharides. (Page S9) Table S1 The jack-knifed classification on single-sphere array for 23 saccharides. (Page S10) Fig. S7 Lower detection concentration at 10 mM of the single-sphere array. (Page S11) Fig. S8 Lower detection concentration at 1 mM of the single-sphere array. (Page S12) Fig. S9 PIL sphere exhibiting great pH robustness within the biological pH range. (Page S12) Fig. S10 Exploring the tolerance of PIL spheres to different conditions. (Page S13) Fig. S11 Exploring the reusability of PIL spheres. (Page S14) Fig. S12 Responses of spheres to sugar alcohols. (Page S15) Fig.
    [Show full text]
  • Lecture 7 - the Calvin Cycle and the Pentose Phosphate Pathway
    Lecture 7 - The Calvin Cycle and the Pentose Phosphate Pathway Chem 454: Regulatory Mechanisms in Biochemistry University of Wisconsin-Eau Claire 1 Introduction The Calvin cycle Text The dark reactions of photosynthesis in green plants Reduces carbon from CO2 to hexose (C6H12O6) Requires ATP for free energy and NADPH as a reducing agent. 2 2 Introduction NADH versus Text NADPH 3 3 Introduction The Pentose Phosphate Pathway Used in all organisms Glucose is oxidized and decarboxylated to produce reduced NADPH Used for the synthesis and degradation of pentoses Shares reactions with the Calvin cycle 4 4 1. The Calvin Cycle Source of carbon is CO2 Text Takes place in the stroma of the chloroplasts Comprises three stages Fixation of CO2 by ribulose 1,5-bisphosphate to form two 3-phosphoglycerate molecules Reduction of 3-phosphoglycerate to produce hexose sugars Regeneration of ribulose 1,5-bisphosphate 5 5 1. Calvin Cycle Three stages 6 6 1.1 Stage I: Fixation Incorporation of CO2 into 3-phosphoglycerate 7 7 1.1 Stage I: Fixation Rubisco: Ribulose 1,5- bisphosphate carboxylase/ oxygenase 8 8 1.1 Stage I: Fixation Active site contains a divalent metal ion 9 9 1.2 Rubisco Oxygenase Activity Rubisco also catalyzes a wasteful oxygenase reaction: 10 10 1.3 State II: Formation of Hexoses Reactions similar to those of gluconeogenesis But they take place in the chloroplasts And use NADPH instead of NADH 11 11 1.3 State III: Regeneration of Ribulose 1,5-Bisphosphosphate Involves a sequence of transketolase and aldolase reactions. 12 12 1.3 State III:
    [Show full text]
  • Conversion of D-Ribulose 5-Phosphate to D-Xylulose 5-Phosphate: New Insights from Structural and Biochemical Studies on Human RPE
    The FASEB Journal • Research Communication Conversion of D-ribulose 5-phosphate to D-xylulose 5-phosphate: new insights from structural and biochemical studies on human RPE Wenguang Liang,*,†,1 Songying Ouyang,*,1 Neil Shaw,* Andrzej Joachimiak,‡ Rongguang Zhang,* and Zhi-Jie Liu*,2 *National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China; †Graduate University of Chinese Academy of Sciences, Beijing, China; and ‡Structural Biology Center, Argonne National Laboratory, Argonne, Illinois, USA ABSTRACT The pentose phosphate pathway (PPP) such as erythrose 4-phosphate, glyceraldehyde 3-phos- confers protection against oxidative stress by supplying phate, and fructose 6-phosphate that are necessary for NADPH necessary for the regeneration of glutathione, the synthesis of aromatic amino acids and production which detoxifies H2O2 into H2O and O2. RPE functions of energy (Fig. 1) (3, 4). In addition, a majority of the in the PPP, catalyzing the reversible conversion of NADPH used by the human body for biosynthetic D-ribulose 5-phosphate to D-xylulose 5-phosphate and is purposes is supplied by the PPP (5). Interestingly, RPE an important enzyme for cellular response against has been shown to protect cells from oxidative stress via oxidative stress. Here, using structural, biochemical, its participation in PPP for the production of NADPH and functional studies, we show that human D-ribulose (6–8). The protection against reactive oxygen species is ؉ 5-phosphate 3-epimerase (hRPE) uses Fe2 for cataly- exerted by NADPH’s ability to reduce glutathione, sis. Structures of the binary complexes of hRPE with which detoxifies H2O2 into H2O (Fig. 1).
    [Show full text]
  • Carbohydrates: Structure and Function
    CARBOHYDRATES: STRUCTURE AND FUNCTION Color index: . Very important . Extra Information. “ STOP SAYING I WISH, START SAYING I WILL” 435 Biochemistry Team *هذا العمل ﻻ يغني عن المصدر المذاكرة الرئيسي • The structure of carbohydrates of physiological significance. • The main role of carbohydrates in providing and storing of energy. • The structure and function of glycosaminoglycans. OBJECTIVES: 435 Biochemistry Team extra information that might help you 1-synovial fluid: - It is a viscous, non-Newtonian fluid found in the cavities of synovial joints. - the principal role of synovial fluid is to reduce friction between the articular cartilage of synovial joints during movement O 2- aldehyde = terminal carbonyl group (RCHO) R H 3- ketone = carbonyl group within (inside) the compound (RCOR’) 435 Biochemistry Team the most abundant organic molecules in nature (CH2O)n Carbohydrates Formula *hydrate of carbon* Function 1-provides important part of energy Diseases caused by disorders of in diet . 2-Acts as the storage form of energy carbohydrate metabolism in the body 3-structural component of cell membrane. 1-Diabetesmellitus. 2-Galactosemia. 3-Glycogen storage disease. 4-Lactoseintolerance. 435 Biochemistry Team Classification of carbohydrates monosaccharides disaccharides oligosaccharides polysaccharides simple sugar Two monosaccharides 3-10 sugar units units more than 10 sugar units Joining of 2 monosaccharides No. of carbon atoms Type of carbonyl by O-glycosidic bond: they contain group they contain - Maltose (α-1, 4)= glucose + glucose -Sucrose (α-1,2)= glucose + fructose - Lactose (β-1,4)= glucose+ galactose Homopolysaccharides Heteropolysaccharides Ketone or aldehyde Homo= same type of sugars Hetero= different types Ketose aldose of sugars branched unBranched -Example: - Contains: - Contains: Examples: aldehyde group glycosaminoglycans ketone group.
    [Show full text]
  • Recent Advances in Trypanosomatid Research: Genome Organization, Expression, Metabolism, Taxonomy and Evolution
    Parasitology Recent advances in trypanosomatid research: genome organization, expression, metabolism, cambridge.org/par taxonomy and evolution 1 2 3,4 5,6 Review Dmitri A. Maslov , Fred R. Opperdoes , Alexei Y. Kostygov , Hassan Hashimi , Julius Lukeš5,6 and Vyacheslav Yurchenko3,5,7 Cite this article: Maslov DA, Opperdoes FR, Kostygov AY, Hashimi H, Lukeš J, Yurchenko V 1Department of Molecular, Cell and Systems Biology, University of California – Riverside, Riverside, California, USA; (2018). Recent advances in trypanosomatid 2de Duve Institute, Université Catholique de Louvain, Brussels, Belgium; 3Life Science Research Centre, Faculty of research: genome organization, expression, 4 metabolism, taxonomy and evolution. Science, University of Ostrava, Ostrava, Czech Republic; Zoological Institute of the Russian Academy of Sciences, 5 Parasitology 1–27. https://doi.org/10.1017/ St. Petersburg, Russia; Biology Centre, Institute of Parasitology, Czech Academy of Sciences, České Budejovice 6 S0031182018000951 (Budweis), Czech Republic; University of South Bohemia, Faculty of Sciences, České Budejovice (Budweis), Czech Republic and 7Martsinovsky Institute of Medical Parasitology, Tropical and Vector Borne Diseases, Sechenov Received: 30 January 2018 University, Moscow, Russia Revised: 23 April 2018 Accepted: 23 April 2018 Abstract Key words: Unicellular flagellates of the family Trypanosomatidae are obligatory parasites of inverte- Gene exchange; kinetoplast; metabolism; molecular and cell biology; taxonomy; brates, vertebrates and plants. Dixenous species are aetiological agents of a number of diseases trypanosomatidae in humans, domestic animals and plants. Their monoxenous relatives are restricted to insects. Because of the high biological diversity, adaptability to dramatically different environmental Author for correspondence: conditions, and omnipresence, these protists have major impact on all biotic communities Vyacheslav Yurchenko, E-mail: vyacheslav.
    [Show full text]
  • PENTOSE PHOSPHATE PATHWAY — Restricted for Students Enrolled in MCB102, UC Berkeley, Spring 2008 ONLY
    Metabolism Lecture 5 — PENTOSE PHOSPHATE PATHWAY — Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY Bryan Krantz: University of California, Berkeley MCB 102, Spring 2008, Metabolism Lecture 5 Reading: Ch. 14 of Principles of Biochemistry, “Glycolysis, Gluconeogenesis, & Pentose Phosphate Pathway.” PENTOSE PHOSPHATE PATHWAY This pathway produces ribose from glucose, and it also generates 2 NADPH. Two Phases: [1] Oxidative Phase & [2] Non-oxidative Phase + + Glucose 6-Phosphate + 2 NADP + H2O Ribose 5-Phosphate + 2 NADPH + CO2 + 2H ● What are pentoses? Why do we need them? ◦ DNA & RNA ◦ Cofactors in enzymes ● Where do we get them? Diet and from glucose (and other sugars) via the Pentose Phosphate Pathway. ● Is the Pentose Phosphate Pathway just about making ribose sugars from glucose? (1) Important for biosynthetic pathways using NADPH, and (2) a high cytosolic reducing potential from NADPH is sometimes required to advert oxidative damage by radicals, e.g., ● - ● O2 and H—O Metabolism Lecture 5 — PENTOSE PHOSPHATE PATHWAY — Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY Two Phases of the Pentose Pathway Metabolism Lecture 5 — PENTOSE PHOSPHATE PATHWAY — Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY NADPH vs. NADH Metabolism Lecture 5 — PENTOSE PHOSPHATE PATHWAY — Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY Oxidative Phase: Glucose-6-P Ribose-5-P Glucose 6-phosphate dehydrogenase. First enzymatic step in oxidative phase, converting NADP+ to NADPH. Glucose 6-phosphate + NADP+ 6-Phosphoglucono-δ-lactone + NADPH + H+ Mechanism. Oxidation reaction of C1 position. Hydride transfer to the NADP+, forming a lactone, which is an intra-molecular ester.
    [Show full text]
  • Table 6. Putative Genes Involved in the Utilization of Carbohydrates in G
    Table 6. Putative genes involved in the utilization of carbohydrates in G. thermodenitrificans NG80-2 genome Carbohydrates* Enzymes Gene ID Glycerol Glycerol Kinase GT1216 Glycerol-3-phosphate dehydrogenase, aerobic GT2089 NAD(P)H-dependent glycerol-3-phosphate dehydrogenase GT2153 Enolase GT3003 2,3-bisphosphoglycerate-independentphosphoglycerate mutase GT3004 Triosephosphate isomerase GT3005 3-phosphoglycerate kinase GT3006 Glyceraldehyde-3-phosphate dehydrogenase GT3007 Phosphoglycerate mutase GT1326 Pyruvate kinase GT2663 L-Arabinose L-arabinose isomerase GT1795 L-ribulokinase GT1796 L-ribulose 5-phosphate 4-epimerase GT1797 D-Ribose Ribokinase GT3174 Transketolase GT1187 Ribose 5-phosphate epimerase GT3316 D-Xylose Xylose kinase GT1756 Xylose isomerase GT1757 D-Galactose Galactokinase GT2086 Galactose-1-phosphate uridyltransferase GT2084 UDP-glucose 4-epimerase GT2085 Carbohydrates* Enzymes Gene ID D-Fructose 1-phosphofructokinase GT1727 Fructose-1,6-bisphosphate aldolase GT1805 Fructose-1,6-bisphosphate aldolase type II GT3331 Triosephosphate isomerase GT3005 D-Mannose Mannnose-6 phospate isomelase GT3398 6-phospho-1-fructokinase GT2664 D-Mannitol Mannitol-1-phosphate dehydrogenase GT1844 N-Acetylglucosamine N-acetylglucosamine-6-phosphate deacetylase GT2205 N-acetylglucosamine-6-phosphate isomerase GT2204 D-Maltose Alpha-1,4-glucosidase GT0528, GT1643 Sucrose Sucrose phosphorylase GT3215 D-Trehalose Alpha-glucosidase GT1643 Glucose kinase GT2381 Inositol Myo-inositol catabolism protein iolC;5-dehydro-2- GT1807 deoxygluconokinase
    [Show full text]
  • A Global Analysis of Enzyme Compartmentalization to Glycosomes
    pathogens Article A Global Analysis of Enzyme Compartmentalization to Glycosomes Hina Durrani 1, Marshall Hampton 2 , Jon N. Rumbley 3 and Sara L. Zimmer 1,* 1 Department of Biomedical Sciences, University of Minnesota Medical School, Duluth Campus, Duluth, MN 55812, USA; [email protected] 2 Mathematics & Statistics Department, University of Minnesota Duluth, Duluth, MN 55812, USA; [email protected] 3 College of Pharmacy, University of Minnesota, Duluth Campus, Duluth, MN 55812, USA; [email protected] * Correspondence: [email protected] Received: 25 March 2020; Accepted: 9 April 2020; Published: 12 April 2020 Abstract: In kinetoplastids, the first seven steps of glycolysis are compartmentalized into a glycosome along with parts of other metabolic pathways. This organelle shares a common ancestor with the better-understood eukaryotic peroxisome. Much of our understanding of the emergence, evolution, and maintenance of glycosomes is limited to explorations of the dixenous parasites, including the enzymatic contents of the organelle. Our objective was to determine the extent that we could leverage existing studies in model kinetoplastids to determine the composition of glycosomes in species lacking evidence of experimental localization. These include diverse monoxenous species and dixenous species with very different hosts. For many of these, genome or transcriptome sequences are available. Our approach initiated with a meta-analysis of existing studies to generate a subset of enzymes with highest evidence of glycosome localization. From this dataset we extracted the best possible glycosome signal peptide identification scheme for in silico identification of glycosomal proteins from any kinetoplastid species. Validation suggested that a high glycosome localization score from our algorithm would be indicative of a glycosomal protein.
    [Show full text]
  • The Microbiota-Produced N-Formyl Peptide Fmlf Promotes Obesity-Induced Glucose
    Page 1 of 230 Diabetes Title: The microbiota-produced N-formyl peptide fMLF promotes obesity-induced glucose intolerance Joshua Wollam1, Matthew Riopel1, Yong-Jiang Xu1,2, Andrew M. F. Johnson1, Jachelle M. Ofrecio1, Wei Ying1, Dalila El Ouarrat1, Luisa S. Chan3, Andrew W. Han3, Nadir A. Mahmood3, Caitlin N. Ryan3, Yun Sok Lee1, Jeramie D. Watrous1,2, Mahendra D. Chordia4, Dongfeng Pan4, Mohit Jain1,2, Jerrold M. Olefsky1 * Affiliations: 1 Division of Endocrinology & Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA. 2 Department of Pharmacology, University of California, San Diego, La Jolla, California, USA. 3 Second Genome, Inc., South San Francisco, California, USA. 4 Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA. * Correspondence to: 858-534-2230, [email protected] Word Count: 4749 Figures: 6 Supplemental Figures: 11 Supplemental Tables: 5 1 Diabetes Publish Ahead of Print, published online April 22, 2019 Diabetes Page 2 of 230 ABSTRACT The composition of the gastrointestinal (GI) microbiota and associated metabolites changes dramatically with diet and the development of obesity. Although many correlations have been described, specific mechanistic links between these changes and glucose homeostasis remain to be defined. Here we show that blood and intestinal levels of the microbiota-produced N-formyl peptide, formyl-methionyl-leucyl-phenylalanine (fMLF), are elevated in high fat diet (HFD)- induced obese mice. Genetic or pharmacological inhibition of the N-formyl peptide receptor Fpr1 leads to increased insulin levels and improved glucose tolerance, dependent upon glucagon- like peptide-1 (GLP-1). Obese Fpr1-knockout (Fpr1-KO) mice also display an altered microbiome, exemplifying the dynamic relationship between host metabolism and microbiota.
    [Show full text]