2.0 2012-05-31 10:22:45 -0600 2015-09-13 12:56:06 -0600 ECMDB00168 M2MDB000069 L-Asparagine Asparagine (Asn) is one of the 20 most common natural amino acids on Earth. It has carboxamide as the side chain's functional group. It is considered a non-essential amino acid. The precursor to asparagine is oxaloacetate. Oxaloacetate is converted to aspartate using a transaminase enzyme. The enzyme transfers the amino group from glutamate to oxaloacetate producing alpha-ketoglutarate and aspartate. The enzyme asparagine synthetase produces asparagine, AMP, glutamate, and pyrophosphate from aspartate, glutamine, and ATP. In the asparagine synthetase reaction, ATP is used to activate aspartate, forming beta-aspartyl-AMP. glutamine donates an ammonium group which reacts with beta-aspartyl-AMP to form asparagine and free AMP. Since the asparagine side chain can make efficient hydrogen bond interactions with the peptide backbone, asparagines are often found near the beginning and end of alpha-helices, and in turn motifs in beta sheets. Its role can be thought as "capping" the hydrogen bond interactions which would otherwise need to be satisfied by the polypeptide backbone. Glutamines have an extra methylene group, have more conformational entropy and thus are less useful in this regard. Asparagine also provides key sites for N-linked glycosylation, modification of the protein chain with the addition of carbohydrate chains. (http://en.wikipedia.org/wiki/Asparagine) &alpha;-aminosuccinamate &alpha;-aminosuccinamic acid (-)-Asparagine (S)-2,4-Diamino-4-oxobutanoate (S)-2,4-Diamino-4-oxobutanoic acid (S)-Asparagine 2,4-Diamino-4-oxo-(S)-Butanoate 2,4-Diamino-4-oxo-(S)-Butanoic acid 2,4-diamino-4-Oxobutanoate, (S)- 2,4-diamino-4-oxobutanoic acid, (S)- 2-Aminosuccinamate 2-Aminosuccinamate, L- 2-Aminosuccinamic acid 2-aminosuccinamic acid, L- a Amminosuccinamate a Amminosuccinamic acid A-Aminosuccinamate A-Aminosuccinamic acid Agedoite Alpha Amminosuccinamate Alpha Amminosuccinamic acid Alpha-Aminosuccinamate Alpha-Aminosuccinamic acid Altheine Asn Asparagine Asparagine acid Asparamide Aspartamate Aspartamic acid Aspartate &beta;-amide Aspartate amide Aspartate b amide Aspartate b-amide Aspartate beta amide Aspartate beta-amide Aspartate β amide Aspartate β-amide Aspartic acid &beta;-amide Aspartic acid amide Aspartic acid b amide Aspartic acid b-amide Aspartic acid beta amide Aspartic acid beta-amide Aspartic acid β amide Aspartic acid β-amide B2,4-(S)-diamino-4-oxo-utanoate B2,4-(S)-diamino-4-oxo-utanoic acid Butanoate, 2,4-diamino-4-oxo-, (S)- Butanoic acid, 2,4-diamino-4-oxo-, (S)- Crystal VI L-&beta;-asparagine L-2,4-Diamino-4-oxobutanoate L-2,4-Diamino-4-oxobutanoic acid L-2-Aminosuccinamate L-2-Aminosuccinamic acid L-Asparagine L-Asparatamine L-Aspartamine L-b-Asparagine L-beta-Asparagine L-β-Asparagine N α Amminosuccinamate α Amminosuccinamic acid α-Aminosuccinamate α-Aminosuccinamic acid C4H8N2O3 132.1179 132.053492132 (2S)-2-amino-3-carbamoylpropanoic acid L-asparagine 70-47-3 N[C@@H](CC(N)=O)C(O)=O InChI=1S/C4H8N2O3/c5-2(4(8)9)1-3(6)7/h2H,1,5H2,(H2,6,7)(H,8,9)/t2-/m0/s1 DCXYFEDJOCDNAF-REOHCLBHSA-N Solid Cytosol Extra-organism Periplasm logp -3.36 ALOGPS logs 0.10 ALOGPS solubility 1.68e+02 g/l ALOGPS melting_point 234-235 oC logp -4.3 ChemAxon pka_strongest_acidic 2 ChemAxon pka_strongest_basic 8.43 ChemAxon iupac (2S)-2-amino-3-carbamoylpropanoic acid ChemAxon average_mass 132.1179 ChemAxon mono_mass 132.053492132 ChemAxon smiles N[C@@H](CC(N)=O)C(O)=O ChemAxon formula C4H8N2O3 ChemAxon inchi InChI=1S/C4H8N2O3/c5-2(4(8)9)1-3(6)7/h2H,1,5H2,(H2,6,7)(H,8,9)/t2-/m0/s1 ChemAxon inchikey DCXYFEDJOCDNAF-REOHCLBHSA-N ChemAxon polar_surface_area 106.41 ChemAxon refractivity 28.35 ChemAxon polarizability 11.68 ChemAxon rotatable_bond_count 3 ChemAxon acceptor_count 4 ChemAxon donor_count 3 ChemAxon physiological_charge 0 ChemAxon formal_charge 0 ChemAxon Alanine, aspartate and glutamate metabolism ec00250 Nitrogen metabolism The biological process of the nitrogen cycle is a complex interplay among many microorganisms catalyzing different reactions, where nitrogen is found in various oxidation states ranging from +5 in nitrate to -3 in ammonia. The ability of fixing atmospheric nitrogen by the nitrogenase enzyme complex is present in restricted prokaryotes (diazotrophs). The other reduction pathways are assimilatory nitrate reduction and dissimilatory nitrate reduction both for conversion to ammonia, and denitrification. Denitrification is a respiration in which nitrate or nitrite is reduced as a terminal electron acceptor under low oxygen or anoxic conditions, producing gaseous nitrogen compounds (N2, NO and N2O) to the atmosphere. Nitrate can be introduced into the cytoplasm through a nitrate:nitrite antiporter NarK or a nitrate / nitrite transporter NarU. Nitrate is then reduced by a Nitrate Reductase resulting in the release of water, an acceptor and a Nitrite. Nitrite can also be introduced into the cytoplasm through a nitrate:nitrite antiporter NarK Nitrite can be reduced a NADPH dependent nitrite reductase resulting in water and NAD and Ammonia. Nitrite can interact with hydrogen ion, ferrocytochrome c through a cytochrome c-552 ferricytochrome resulting in the release of ferricytochrome c, water and ammonia Another process by which ammonia is produced is by a reversible reaction of hydroxylamine with a reduced acceptor through a hydroxylamine reductase resulting in an acceptor, water and ammonia. Water and carbon dioxide react through a carbonate dehydratase resulting in carbamic acid. This compound reacts spontaneously with hydrogen ion resulting in the release of carbon dioxide and ammonia. Carbon dioxide can interact with water through a carbonic anhydrase resulting in hydrogen carbonate. This compound interacts with cyanate and hydrogen ion through a cyanate hydratase resulting in a carbamic acid. Ammonia can be metabolized by reacting with L-glutamine and ATP driven glutamine synthetase resulting in ADP, phosphate and L-glutamine. The latter compound reacts with oxoglutaric acid and hydrogen ion through a NADPH dependent glutamate synthase resulting in the release of NADP and L-glutamic acid. L-glutamic acid reacts with water through a NADP-specific glutamate dehydrogenase resulting in the release of oxoglutaric acid, NADPH, hydrogen ion and ammonia. PW000755 ec00910 Metabolic Cyanoamino acid metabolism ec00460 Aminoacyl-tRNA biosynthesis ec00970 Metabolic pathways eco01100 Asparagine biosynthesis L-asparagine is synthesized in E. coli from L-aspartate by either of two reactions, utilizing either L-glutamine or ammonia as the amino group donor. Both reactions are ATP driven and yield AMP and pyrophosphate. The first reaction is catalyzed only by asparagine synthetase B, while the second reaction is catalyzed by both asparagine synthetase A and asparagine synthetase B, The only known role of asparagine in the metabolism of E. coli is as a constituent of protein. PW000813 Metabolic Aspartate metabolism Aspartate (seen in the center) is synthesized from and degraded to oxaloacetate , an intermediate of the TCA cycle, by a reversible transamination reaction with glutamate. As shown here, AspC is the principal transaminase that catalyzes this reaction, but TyrB also catalyzes it. Null mutations in aspC do not confer aspartate auxotrophy; null mutations in both aspC and tyrB do. Aspartate is a constituent of proteins and participates in several other biosyntheses as shown here( NAD biosynthesis and Beta-Alanine Metabolism . Approximately 27 percent of the cell's nitrogen flows through aspartate Aspartate can be synthesized from fumaric acid through a aspartate ammonia lyase. Aspartate also participates in the synthesis of L-asparagine through two different methods, either through aspartate ammonia ligase or asparagine synthetase B. Aspartate is also a precursor of fumaric acid. Again it has two possible ways of synthesizing it. First set of reactions follows an adenylo succinate synthetase that yields adenylsuccinic acid and then adenylosuccinate lyase in turns leads to fumaric acid. The second way is through argininosuccinate synthase that yields argininosuccinic acid and then argininosuccinate lyase in turns leads to fumaric acid PW000787 Metabolic inner membrane transport list of inner membrane transport complexes, transporting compounds from the periplasmic space to the cytosol This pathway should be updated regularly with the new inner membrae transports added PW000786 Metabolic tRNA Charging 2 This pathway groups together all E. coli tRNA charging reactions. PW000803 Metabolic tRNA charging This pathway groups together all E. coli tRNA charging reactions. PW000799 Metabolic asparagine biosynthesis I ASPARAGINE-BIOSYNTHESIS tRNA charging TRNA-CHARGING-PWY asparagine biosynthesis II ASPARAGINESYN-PWY superpathway of aspartate and asparagine biosynthesis; interconversion of aspartate and asparagine ASPASN-PWY asparagine degradation I ASPARAGINE-DEG1-PWY Specdb::CMs 401 Specdb::CMs 402 Specdb::CMs 403 Specdb::CMs 404 Specdb::CMs 1192 Specdb::CMs 1254 Specdb::CMs 1258 Specdb::CMs 1289 Specdb::CMs 1309 Specdb::CMs 1338 Specdb::CMs 1566 Specdb::CMs 2895 Specdb::CMs 30028 Specdb::CMs 30199 Specdb::CMs 30302 Specdb::CMs 30589 Specdb::CMs 30736 Specdb::CMs 30882 Specdb::CMs 31040 Specdb::CMs 31041 Specdb::CMs 31042 Specdb::CMs 31043 Specdb::CMs 31044 Specdb::CMs 31045 Specdb::CMs 31046 Specdb::NmrOneD 1132 Specdb::NmrOneD 1186 Specdb::NmrOneD 4803 Specdb::NmrOneD 142670 Specdb::NmrOneD 142671 Specdb::NmrOneD 142672 Specdb::NmrOneD 142673 Specdb::NmrOneD 142674 Specdb::NmrOneD 142675 Specdb::NmrOneD 142676 Specdb::NmrOneD 142677 Specdb::NmrOneD 142678 Specdb::NmrOneD 142679 Specdb::NmrOneD 142680 Specdb::NmrOneD 142681 Specdb::NmrOneD 142682 Specdb::NmrOneD 142683 Specdb::NmrOneD 142684 Specdb::NmrOneD 142685 Specdb::NmrOneD 142686 Specdb::NmrOneD 142687 Specdb::NmrOneD 142688 Specdb::NmrOneD 142689 Specdb::NmrOneD 166511 Specdb::NmrOneD 166570 Specdb::MsMs 266 Specdb::MsMs 267 Specdb::MsMs 268 Specdb::MsMs 3281 Specdb::MsMs 3282 Specdb::MsMs 3283 Specdb::MsMs 3284 Specdb::MsMs 3285 Specdb::MsMs 3286 Specdb::MsMs 3287 Specdb::MsMs 3288 Specdb::MsMs 3289 Specdb::MsMs 3290 Specdb::MsMs 3291 Specdb::MsMs 3292 Specdb::MsMs 3293 Specdb::MsMs 3294 Specdb::MsMs 3295 Specdb::MsMs 3296 Specdb::MsMs 3297 Specdb::MsMs 3298 Specdb::MsMs 3299 Specdb::MsMs 3300 Specdb::MsMs 3301 Specdb::MsMs 3302 Specdb::NmrTwoD 980 Specdb::NmrTwoD 1183 HMDB00168 6267 6031 C00152 17196 ASN ASN_LFZW Asn Keseler, I. 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Faming Zhuanli Shenqing Gongkai Shuomingshu (2005), 8 pp. http://hmdb.ca/system/metabolites/msds/000/000/118/original/HMDB00168.pdf?1358463305 L-asparaginase 2 P00805 ASPG2_ECOLI ansB http://ecmdb.ca/proteins/P00805.xml Aspartate--ammonia ligase P00963 ASNA_ECOLI asnA http://ecmdb.ca/proteins/P00963.xml Asparaginyl-tRNA synthetase P0A8M0 SYN_ECOLI asnS http://ecmdb.ca/proteins/P0A8M0.xml L-asparaginase 1 P0A962 ASPG1_ECOLI ansA http://ecmdb.ca/proteins/P0A962.xml Asparagine synthetase B [glutamine-hydrolyzing] P22106 ASNB_ECOLI asnB http://ecmdb.ca/proteins/P22106.xml Isoaspartyl peptidase P37595 IAAA_ECOLI iaaA http://ecmdb.ca/proteins/P37595.xml Uncharacterized amino-acid ABC transporter ATP-binding protein yecC P37774 YECC_ECOLI yecC http://ecmdb.ca/proteins/P37774.xml Inner membrane amino-acid ABC transporter permease protein yecS P0AFT2 YECS_ECOLI yecS http://ecmdb.ca/proteins/P0AFT2.xml L-asparagine permease P77610 ANSP_ECOLI ansP http://ecmdb.ca/proteins/P77610.xml Outer membrane protein N P77747 OMPN_ECOLI ompN http://ecmdb.ca/proteins/P77747.xml Outer membrane pore protein E P02932 PHOE_ECOLI phoE http://ecmdb.ca/proteins/P02932.xml Outer membrane protein F P02931 OMPF_ECOLI ompF http://ecmdb.ca/proteins/P02931.xml Outer membrane protein C P06996 OMPC_ECOLI ompC http://ecmdb.ca/proteins/P06996.xml L-Asparagine + Water > L-Aspartic acid + Ammonium L-Aspartic acid + Adenosine triphosphate + L-Glutamine + Water > Adenosine monophosphate + L-Asparagine + L-Glutamate + Hydrogen ion + Pyrophosphate R00578 ASNSYNB-RXN L-Asparagine + Adenosine triphosphate + tRNA(Asn) + tRNA(Asn) <> Adenosine monophosphate + L-Asparaginyl-tRNA(Asn) + Pyrophosphate + L-Asparaginyl-tRNA(Asn) R03648 L-Aspartic acid + Adenosine triphosphate + Ammonium > Adenosine monophosphate + L-Asparagine + Hydrogen ion + Pyrophosphate Adenosine triphosphate + L-Aspartic acid + Ammonia <> Adenosine monophosphate + Pyrophosphate + L-Asparagine R00483 L-Asparagine + Water <> L-Aspartic acid + Ammonia R00485 Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water <> Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-Glutamate R00578 Adenosine triphosphate + L-Asparagine + tRNA(Asn) <> Adenosine monophosphate + Pyrophosphate + L-Asparaginyl-tRNA(Asn) R03648 Ammonia + L-Aspartic acid + Adenosine triphosphate > L-Asparagine + Pyrophosphate + Adenosine monophosphate R00483 ASNSYNA-RXN L-Asparagine + Water > Hydrogen ion + L-Aspartic acid + Ammonia R00485 ASPARAGHYD-RXN gly-asn + Water > Glycine + L-Asparagine RXN0-6982 Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water > Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-Glutamate L-Asparagine + Water > L-Aspartic acid + Ammonia Adenosine triphosphate + L-Asparagine + tRNA(Asn) > Adenosine monophosphate + Pyrophosphate + L-asparaginyl-tRNA(Asn) Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water + Ammonia <> Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-Glutamate R00578 Adenosine triphosphate + L-Aspartic acid + Ammonia + L-Aspartic acid > Adenosine monophosphate + L-Asparagine + Pyrophosphate + L-Asparagine PW_R002642 Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water + L-Aspartic acid > Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-Glutamic acid + L-Asparagine + L-Glutamate PW_R002643 L-Asparagine + Adenosine triphosphate + Hydrogen ion + tRNA(Asn) + L-Asparagine > Pyrophosphate + Adenosine monophosphate + L-asparaginyl-tRNA(Asn) PW_R002838 L-Aspartic acid + Water + Adenosine triphosphate + L-Glutamine + L-Aspartic acid > L-Asparagine + Hydrogen ion + Adenosine monophosphate + L-Glutamic acid + Pyrophosphate + L-Asparagine + L-Glutamate PW_R002887 L-Aspartic acid + Adenosine triphosphate + Ammonium + L-Aspartic acid > L-Asparagine + Adenosine monophosphate + Pyrophosphate + Hydrogen ion + L-Asparagine PW_R002888 L-Asparagine + Water + L-Asparagine > L-Aspartic acid + Ammonium + L-Aspartic acid PW_R003388 L-Asparagine + Water <> L-Aspartic acid + Ammonia L-Asparagine + Water > Hydrogen ion + L-Aspartic acid + Ammonia Gutnick minimal complete medium (4.7 g/L KH2PO4; 13.5 g/L K2HPO4; 1 g/L K2SO4; 0.1 g/L MgSO4-7H2O; 10 mM NH4Cl) with 4 g/L glucose Shake flask and filter culture 511.0 uM 0.0 37 oC K12 NCM3722 Mid-Log Phase 2044000 0 Bennett, B. D., Kimball, E. H., Gao, M., Osterhout, R., Van Dien, S. J., Rabinowitz, J. D. (2009). "Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli." Nat Chem Biol 5:593-599. 19561621 Gutnick minimal complete medium (4.7 g/L KH2PO4; 13.5 g/L K2HPO4; 1 g/L K2SO4; 0.1 g/L MgSO4-7H2O; 10 mM NH4Cl) with 4 g/L glycerol Shake flask and filter culture 970.0 uM 0.0 37 oC K12 NCM3722 Mid-Log Phase 3880000 0 Bennett, B. D., Kimball, E. H., Gao, M., Osterhout, R., Van Dien, S. J., Rabinowitz, J. D. (2009). "Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli." Nat Chem Biol 5:593-599. 19561621 Gutnick minimal complete medium (4.7 g/L KH2PO4; 13.5 g/L K2HPO4; 1 g/L K2SO4; 0.1 g/L MgSO4-7H2O; 10 mM NH4Cl) with 4 g/L acetate Shake flask and filter culture 540.0 uM 0.0 37 oC K12 NCM3722 Mid-Log Phase 2160000 0 Bennett, B. D., Kimball, E. H., Gao, M., Osterhout, R., Van Dien, S. J., Rabinowitz, J. D. (2009). "Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli." Nat Chem Biol 5:593-599. 19561621 48 mM Na2HPO4, 22 mM KH2PO4, 10 mM NaCl, 45 mM (NH4)2SO4, supplemented with 1 mM MgSO4, 1 mg/l thiamine·HCl, 5.6 mg/l CaCl2, 8 mg/l FeCl3, 1 mg/l MnCl2·4H2O, 1.7 mg/l ZnCl2, 0.43 mg/l CuCl2·2H2O, 0.6 mg/l CoCl2·2H2O and 0.6 mg/l Na2MoO4·2H2O. 4 g/L Gluco Bioreactor, pH controlled, O2 and CO2 controlled, dilution rate: 0.2/h 103.0 uM 0.0 37 oC BW25113 Stationary Phase, glucose limited 412000 0 Ishii, N., Nakahigashi, K., Baba, T., Robert, M., Soga, T., Kanai, A., Hirasawa, T., Naba, M., Hirai, K., Hoque, A., Ho, P. Y., Kakazu, Y., Sugawara, K., Igarashi, S., Harada, S., Masuda, T., Sugiyama, N., Togashi, T., Hasegawa, M., Takai, Y., Yugi, K., Arakawa, K., Iwata, N., Toya, Y., Nakayama, Y., Nishioka, T., Shimizu, K., Mori, H., Tomita, M. (2007). "Multiple high-throughput analyses monitor the response of E. coli to perturbations." Science 316:593-597. 17379776 Luria-Bertani (LB) media Shake flask 0.73 uM true 0.05 37 oC BL21 DE3 Stationary phase cultures (overnight culture) 2935 191 Lin, Z., Johnson, L. C., Weissbach, H., Brot, N., Lively, M. O., Lowther, W. T. (2007). "Free methionine-(R)-sulfoxide reductase from Escherichia coli reveals a new GAF domain function." Proc Natl Acad Sci U S A 104:9597-9602. 17535911