2.02012-05-31 10:22:45 -06002015-09-13 12:56:06 -0600ECMDB00168M2MDB000069L-AsparagineAsparagine (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)α-aminosuccinamateα-aminosuccinamic acid(-)-Asparagine(S)-2,4-Diamino-4-oxobutanoate(S)-2,4-Diamino-4-oxobutanoic acid(S)-Asparagine2,4-Diamino-4-oxo-(S)-Butanoate2,4-Diamino-4-oxo-(S)-Butanoic acid2,4-diamino-4-Oxobutanoate, (S)-2,4-diamino-4-oxobutanoic acid, (S)-2-Aminosuccinamate2-Aminosuccinamate, L-2-Aminosuccinamic acid2-aminosuccinamic acid, L-a Amminosuccinamatea Amminosuccinamic acidA-AminosuccinamateA-Aminosuccinamic acidAgedoiteAlpha AmminosuccinamateAlpha Amminosuccinamic acidAlpha-AminosuccinamateAlpha-Aminosuccinamic acidAltheineAsnAsparagineAsparagine acidAsparamideAspartamateAspartamic acidAspartate β-amideAspartate amideAspartate b amideAspartate b-amideAspartate beta amideAspartate beta-amideAspartate β amideAspartate β-amideAspartic acid β-amideAspartic acid amideAspartic acid b amideAspartic acid b-amideAspartic acid beta amideAspartic acid beta-amideAspartic acid β amideAspartic acid β-amideB2,4-(S)-diamino-4-oxo-utanoateB2,4-(S)-diamino-4-oxo-utanoic acidButanoate, 2,4-diamino-4-oxo-, (S)-Butanoic acid, 2,4-diamino-4-oxo-, (S)-Crystal VIL-β-asparagineL-2,4-Diamino-4-oxobutanoateL-2,4-Diamino-4-oxobutanoic acidL-2-AminosuccinamateL-2-Aminosuccinamic acidL-AsparagineL-AsparatamineL-AspartamineL-b-AsparagineL-beta-AsparagineL-β-AsparagineNα Amminosuccinamateα Amminosuccinamic acidα-Aminosuccinamateα-Aminosuccinamic acidC4H8N2O3132.1179132.053492132(2S)-2-amino-3-carbamoylpropanoic acidL-asparagine70-47-3N[C@@H](CC(N)=O)C(O)=OInChI=1S/C4H8N2O3/c5-2(4(8)9)1-3(6)7/h2H,1,5H2,(H2,6,7)(H,8,9)/t2-/m0/s1DCXYFEDJOCDNAF-REOHCLBHSA-NSolidCytosolExtra-organismPeriplasmlogp-3.36logs0.10solubility1.68e+02 g/lmelting_point234-235 oClogp-4.3pka_strongest_acidic2pka_strongest_basic8.43iupac(2S)-2-amino-3-carbamoylpropanoic acidaverage_mass132.1179mono_mass132.053492132smilesN[C@@H](CC(N)=O)C(O)=OformulaC4H8N2O3inchiInChI=1S/C4H8N2O3/c5-2(4(8)9)1-3(6)7/h2H,1,5H2,(H2,6,7)(H,8,9)/t2-/m0/s1inchikeyDCXYFEDJOCDNAF-REOHCLBHSA-Npolar_surface_area106.41refractivity28.35polarizability11.68rotatable_bond_count3acceptor_count4donor_count3physiological_charge0formal_charge0Alanine, aspartate and glutamate metabolismec00250Nitrogen 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.
PW000755ec00910MetabolicCyanoamino acid metabolismec00460Aminoacyl-tRNA biosynthesisec00970Metabolic pathwayseco01100Asparagine biosynthesisL-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. PW000813MetabolicAspartate metabolismAspartate (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
PW000787Metabolicinner membrane transportlist 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 addedPW000786MetabolictRNA Charging 2This pathway groups together all E. coli tRNA charging reactions.PW000803MetabolictRNA chargingThis pathway groups together all E. coli tRNA charging reactions.PW000799Metabolicasparagine biosynthesis IASPARAGINE-BIOSYNTHESIStRNA chargingTRNA-CHARGING-PWYasparagine biosynthesis IIASPARAGINESYN-PWYsuperpathway of aspartate and asparagine biosynthesis; interconversion of aspartate and asparagineASPASN-PWYasparagine degradation IASPARAGINE-DEG1-PWYSpecdb::CMs401Specdb::CMs402Specdb::CMs403Specdb::CMs404Specdb::CMs1192Specdb::CMs1254Specdb::CMs1258Specdb::CMs1289Specdb::CMs1309Specdb::CMs1338Specdb::CMs1566Specdb::CMs2895Specdb::CMs30028Specdb::CMs30199Specdb::CMs30302Specdb::CMs30589Specdb::CMs30736Specdb::CMs30882Specdb::CMs31040Specdb::CMs31041Specdb::CMs31042Specdb::CMs31043Specdb::CMs31044Specdb::CMs31045Specdb::CMs31046Specdb::NmrOneD1132Specdb::NmrOneD1186Specdb::NmrOneD4803Specdb::NmrOneD142670Specdb::NmrOneD142671Specdb::NmrOneD142672Specdb::NmrOneD142673Specdb::NmrOneD142674Specdb::NmrOneD142675Specdb::NmrOneD142676Specdb::NmrOneD142677Specdb::NmrOneD142678Specdb::NmrOneD142679Specdb::NmrOneD142680Specdb::NmrOneD142681Specdb::NmrOneD142682Specdb::NmrOneD142683Specdb::NmrOneD142684Specdb::NmrOneD142685Specdb::NmrOneD142686Specdb::NmrOneD142687Specdb::NmrOneD142688Specdb::NmrOneD142689Specdb::NmrOneD166511Specdb::NmrOneD166570Specdb::MsMs266Specdb::MsMs267Specdb::MsMs268Specdb::MsMs3281Specdb::MsMs3282Specdb::MsMs3283Specdb::MsMs3284Specdb::MsMs3285Specdb::MsMs3286Specdb::MsMs3287Specdb::MsMs3288Specdb::MsMs3289Specdb::MsMs3290Specdb::MsMs3291Specdb::MsMs3292Specdb::MsMs3293Specdb::MsMs3294Specdb::MsMs3295Specdb::MsMs3296Specdb::MsMs3297Specdb::MsMs3298Specdb::MsMs3299Specdb::MsMs3300Specdb::MsMs3301Specdb::MsMs3302Specdb::NmrTwoD980Specdb::NmrTwoD1183HMDB0016862676031C0015217196ASNASN_LFZWAsnKeseler, 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?1358463305L-asparaginase 2P00805ASPG2_ECOLIansBhttp://ecmdb.ca/proteins/P00805.xmlAspartate--ammonia ligaseP00963ASNA_ECOLIasnAhttp://ecmdb.ca/proteins/P00963.xmlAsparaginyl-tRNA synthetaseP0A8M0SYN_ECOLIasnShttp://ecmdb.ca/proteins/P0A8M0.xmlL-asparaginase 1P0A962ASPG1_ECOLIansAhttp://ecmdb.ca/proteins/P0A962.xmlAsparagine synthetase B [glutamine-hydrolyzing]P22106ASNB_ECOLIasnBhttp://ecmdb.ca/proteins/P22106.xmlIsoaspartyl peptidaseP37595IAAA_ECOLIiaaAhttp://ecmdb.ca/proteins/P37595.xmlUncharacterized amino-acid ABC transporter ATP-binding protein yecCP37774YECC_ECOLIyecChttp://ecmdb.ca/proteins/P37774.xmlInner membrane amino-acid ABC transporter permease protein yecSP0AFT2YECS_ECOLIyecShttp://ecmdb.ca/proteins/P0AFT2.xmlL-asparagine permeaseP77610ANSP_ECOLIansPhttp://ecmdb.ca/proteins/P77610.xmlOuter membrane protein NP77747OMPN_ECOLIompNhttp://ecmdb.ca/proteins/P77747.xmlOuter membrane pore protein EP02932PHOE_ECOLIphoEhttp://ecmdb.ca/proteins/P02932.xmlOuter membrane protein FP02931OMPF_ECOLIompFhttp://ecmdb.ca/proteins/P02931.xmlOuter membrane protein CP06996OMPC_ECOLIompChttp://ecmdb.ca/proteins/P06996.xmlL-Asparagine + Water > L-Aspartic acid + AmmoniumL-Aspartic acid + Adenosine triphosphate + L-Glutamine + Water > Adenosine monophosphate + L-Asparagine + L-Glutamate + Hydrogen ion + PyrophosphateR00578ASNSYNB-RXNL-Asparagine + Adenosine triphosphate + tRNA(Asn) + tRNA(Asn) <> Adenosine monophosphate + L-Asparaginyl-tRNA(Asn) + Pyrophosphate + L-Asparaginyl-tRNA(Asn)R03648L-Aspartic acid + Adenosine triphosphate + Ammonium > Adenosine monophosphate + L-Asparagine + Hydrogen ion + PyrophosphateAdenosine triphosphate + L-Aspartic acid + Ammonia <> Adenosine monophosphate + Pyrophosphate + L-AsparagineR00483L-Asparagine + Water <> L-Aspartic acid + AmmoniaR00485Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water <> Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-GlutamateR00578Adenosine triphosphate + L-Asparagine + tRNA(Asn) <> Adenosine monophosphate + Pyrophosphate + L-Asparaginyl-tRNA(Asn)R03648Ammonia + L-Aspartic acid + Adenosine triphosphate > L-Asparagine + Pyrophosphate + Adenosine monophosphateR00483ASNSYNA-RXNL-Asparagine + Water > Hydrogen ion + L-Aspartic acid + AmmoniaR00485ASPARAGHYD-RXNgly-asn + Water > Glycine + L-AsparagineRXN0-6982Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water > Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-GlutamateL-Asparagine + Water > L-Aspartic acid + AmmoniaAdenosine 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-GlutamateR00578 Adenosine triphosphate + L-Aspartic acid + Ammonia + L-Aspartic acid > Adenosine monophosphate + L-Asparagine + Pyrophosphate + L-AsparaginePW_R002642Adenosine triphosphate + L-Aspartic acid + L-Glutamine + Water + L-Aspartic acid > Adenosine monophosphate + Pyrophosphate + L-Asparagine + L-Glutamic acid + L-Asparagine + L-GlutamatePW_R002643L-Asparagine + Adenosine triphosphate + Hydrogen ion + tRNA(Asn) + L-Asparagine > Pyrophosphate + Adenosine monophosphate + L-asparaginyl-tRNA(Asn)PW_R002838L-Aspartic acid + Water + Adenosine triphosphate + L-Glutamine + L-Aspartic acid > L-Asparagine + Hydrogen ion + Adenosine monophosphate + L-Glutamic acid + Pyrophosphate + L-Asparagine + L-GlutamatePW_R002887L-Aspartic acid + Adenosine triphosphate + Ammonium + L-Aspartic acid > L-Asparagine + Adenosine monophosphate + Pyrophosphate + Hydrogen ion + L-AsparaginePW_R002888L-Asparagine + Water + L-Asparagine > L-Aspartic acid + Ammonium + L-Aspartic acidPW_R003388L-Asparagine + Water <> L-Aspartic acid + AmmoniaL-Asparagine + Water > Hydrogen ion + L-Aspartic acid + AmmoniaGutnick 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 glucoseShake flask and filter culture511.0uM0.037 oCK12 NCM3722Mid-Log Phase20440000Bennett, 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 5:593-599.19561621Gutnick 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 glycerolShake flask and filter culture970.0uM0.037 oCK12 NCM3722Mid-Log Phase38800000Bennett, 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 5:593-599.19561621Gutnick 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 acetateShake flask and filter culture540.0uM0.037 oCK12 NCM3722Mid-Log Phase21600000Bennett, 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 5:593-599.1956162148 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 GlucoBioreactor, pH controlled, O2 and CO2 controlled, dilution rate: 0.2/h103.0uM0.037 oCBW25113Stationary Phase, glucose limited4120000Ishii, 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 316:593-597.17379776Luria-Bertani (LB) mediaShake flask0.73uMtrue0.0537 oCBL21 DE3Stationary phase cultures (overnight culture)2935191Lin, 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