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Faculty Publications from the Center for Plant Science Innovation Plant Science Innovation, Center for

2005

Folate Synthesis and in Plants and Prospects For Biofortification

Gilles J. C. Basset University of Nebraska-Lincoln, [email protected]

Eoin P. Quinlivan University of Florida

Jesse F. Gregory III University of Florida

Andrew D. Hanson University of Florida, [email protected]

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Basset, Gilles J. C.; Quinlivan, Eoin P.; Gregory, Jesse F. III; and Hanson, Andrew D., " Synthesis and Metabolism in Plants and Prospects For Biofortification" (2005). Faculty Publications from the Center for Plant Science Innovation. 72. https://digitalcommons.unl.edu/plantscifacpub/72

This Article is brought to you for free and open access by the Plant Science Innovation, Center for at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Faculty Publications from the Center for Plant Science Innovation by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Reproduced from Crop Science. Published by Crop Science Society of America. All copyrights reserved. ecin.Fltsaetiatt oeue,cmoe of composed , molecules, tripartite are transfer one-carbon reactions. mediate that for cofactors ae oeue ihashort a with molecules lated 2001). Roje, and and (Hanson acids photorespiration and amino for- mation lignin acids, plants—in in nucleic and—specifically of pantothenate, biosynthesis includ- the in- functions metabolic ing are key of folates range the wide units, a of in one-carbon volved of N-10 donors moiety, and pterin carriers the of N-5 p at attached levels oxidation are various at units One-carbon 1). (Fig. ties ayae oae r h rfre usrtsfrmost for substrates preferred the polyglu- are These folates glutamate. first tamylated the to attached residues atcmons(sobt,guahoe n ybind- by antioxi- and by glutathione) stabilized (ascorbate, be compounds may dant folates vivo, in However, o -96.Rcie oebr2 03 ypsa *Corresponding grant ([email protected]). Symposia. by 2003. author 2, Series and November Journal Received Foundation Foundation. R-09861. Sr. Science no. National Griffin, the was C.V. from by work the MCB-0129944 Station, Experiment from This Agricultural endowment 32611. Florida an the FL by Nutrition Gainesville, part Human in Florida, and E.P. supported of Science Dep., Food University Sciences III, Dep., Horticultural Gregory J.F. Hanson, and A.D. Quinlivan and Basset G.J.C. oxidative to all, and pterin particularly are the between folates unstable, cleavage Natural degrees, 2000). varying al., to et monogluta- (Scott prefer forms animals—generally mate least, in transporters—at known folate al., hand, those et other the Scott On 2000; 2000). (Cossins, enzymes folate-dependent T engineering. enhance metabolic to by strategies accumulation and possible synthesis consider folate also plant We trans- including catabolism. elucidated, be and to yet port sec- have out that point metabolism and folate folate of compartmentation plant tors unique the its defining and in pathway progress outline synthesis (bioforti- we content review, this folate In enhanced fication). with plants much engineering is there in defects Consequently, interest diseases. tube serious vascular neural has and cancers, of and infants, world risk folates in the increased of in including lack deficiency consequences, However, health foods. common most plant the from them is primarily acquire must diet, and the novo de from folates microorgan- synthesize and cannot plants humans Unlike isms, nucleic pantothenate. of and biosynthesis acids, the amino for required acids, are and organisms living most ct ta. 00 n hspoesi rmtdb light. by promoted is process this and 2000) al., et Scott 7 .SgeR. aio,W 31 USA 53711 WI Madison, Rd., Segoe S. 677 (2005). 45:449–453 © Sci. Crop in Published B oey rbigdbtenteto smajor As two. the between bridged or moiety, ABA rpSineSceyo mrc m America of Society Science Crop aua oae cu rdmnnl spolyglutamy- as predominantly occur folates Natural oae r seta oatr o n-abntase ecin in reactions transfer one-carbon for cofactors essential are Folates 1 hs uhr otiue qal oti article. this to equally contributed authors These monly etrahydrofolate oaeSnhssadMtbls nPat n rset o Biofortification For Prospects and Plants in Metabolism and Synthesis Folate p aioezae( -aminobenzoate rue ne h aeof name the under grouped ilsJ .Basset C. J. Gilles ABSTRACT TF n t derivatives—com- its and (THF) p B) n ltmt moie- glutamate and ABA), ␥ lne hi fglutamyl of chain -linked p 1 onP Quinlivan P. Eoin , B oeis(i.1; (Fig. moieties ABA Published onlineJanuary31,2005 folates aevital —are 1 es .GeoyII n nrwD Hanson* D. Andrew and III, Gregory F. Jesse , 449 rm nteUAwt ytei oi cdhv very have acid folic pro- synthetic Fortification with 2000). USA al., the et in (Scott grams defects tube neu- and ral disease, cardiovascular cancer, as such diseases fetvl erae h niec fflt deficiency folate of incidence the decreased effectively etto nustesm eurn ot erafter year costs recurrent same the supple- also or incurs is fortification It mentation since malnutrition. approach, folate cost-effective global a addressing in alterna- rational tive a with folate therefore is diet enhanced (biofortification) with the content plants of Engineering pills. supplementation acid is folic as in 2003). implement countries, to excessive Gregory, poor difficult of is and fortification cases food Quinlivan Moreover, in 2000; drawbacks (Lucock, health intake have may but lnsi rc uniis yial,patflt contents in folate present plant are Typically, quantities. Folates trace 2000). in al., plants et (Scott proteins to ing eri onr fe onr Bus 2002). (Bouis, country after country in year id(unia ta. 2003). al., et (Quinlivan fied aea nbcei,adi snwams completely 2). (Fig. almost plants from now characterized eight and pathway, is cloned the been to have it specific enzymes and 10 Of , elucidated. in as same nadto,arato nqet lnstereversible plants—the of to unique esterification reaction a addition, In intake be folate Low to 2000). al., supposed et Scott are 2000; variety (Lucock, good and abundance food nations, where population, industrialized world’s the 2000). of including—surprisingly—many most al., for et suboptimal folate is (Scott that intake shows folates single evidence epidemiological of the Compelling are source foods important plant most humans, their on For entirely supply. depend dietary and novo Cossins, de folates 1975; synthesize al., et (Shin plants in 2000). folate usually major is where the seeds, 5-methyl-THF of to exception predominates, the 0.05 5-formyl-THF With and 2001). (roots), al., 2 et bonnet to g 0.2 nmol (leaves), (fruits) 5 0.5 to 2 from range ino TCIatvt mn uclua fractions subcellular Abbreviations: among activity GTPCHI of distribu- the quantifying tion by this confirm to enzymes. attempts cytosolic Our are hence and GTPCHIs peptides al., plant targeting et lack that suggests (Basset evidence organisms homolo- Genomic other each 2002). from domains, GTPCHIs similar to two 2) gous of Fig. (GTPCHI, formed I is cyclohydrolase which GTP unusual an by Branch Pterin stogtt otiuet h tooyo eea major several of etiology the to contribute to thought is ae TCI GTP GTPCHI, lase; t;DN iyrnotrn PH folylpoly- FPGH, dihydroneopterin; DHN, ate; h ln oaesnhssptwyi setal the essentially is pathway synthesis folate plant The niepat n irognss nml cannot animals microorganisms, and plants Unlike h is tpo h ln trnbac scatalyzed is branch pterin plant the of step first The oaeBoytei n Compartmentation and Biosynthesis Folate p ABA, p ylhdoaeI H,tetrahydrofolate. THF, I; cyclohydrolase B ihguoehsbe identi- been glucose—has with ABA

Ϫ p 1 aioezae D,aminodeoxychoris- ADC, -aminobenzoate; rs egt(osn,20;Gam- 2000; (Cossins, weight fresh ␥ guaaehydro- -glutamate Reproduced from Crop Science. Published by Crop Science Society of America. All copyrights reserved. odmntaeta hsognlehsn detectable no has organelle ( this activity able GTPCHI that been have demonstrate is we to activity mitochondria, purified degradative from this absent de- as fractions (DHN) However, dihydroneopterin chloroplast triphosphate. and interfering reaction the an cytosolic grading of the because in failed activity have leaves tomato of C1-substituted its and tetrahydrofolate of structures Chemical 1. Fig. 450 ini lnsi nnw.Termiigpopaeis phosphate remaining The reac- unknown. the of is nature sug- plants The been in 1995). tion also al., et has Saizieu process (De al., specific gested chemical et a a Lee although by 1974; 1999), Brown, mediated and probably (Suzuki most pyrophosphatase is reaction this w tgs h is sls fprpopae nbacteria, in pyrophosphate; of loss is first in The occurs stages. two 2) de- (Fig. triphosphate The DHN extramitochondrial. of phosphorylation is GTPCHI that confirms aeto mate p plants. from has cloned This been 2). yet (Fig. DHN not aldolase of DHN chain by side mediated three-carbon is the of cleavage the ria, hncevdofb oseii hshts.I bacte- In phosphatase. nonspecific a by off cleaved then 92 Fg ) n tapaslkl htteei iia,pat,weesi sctslci te raim.Folates cytosolic organisms. most other a or in by cytosolic all al., is ester that it et glucose whereas and that plasts, glucosyltransferase its shown (Basset to been plants converted has It reversibly in it enzyme Interestingly, 2004). unidentified, similar, a is yet there that likely as appears it and 2), (Fig. 1992) to aromatized subsequently al., is et p ADC (Basset plastidial bacteria, is In synthase 2004). ADC show plant experiments the localization do- that Subcellular tandem 2004). (Basset with al., subunits bacterial et protein two the fusion to homologous bipartite plant mains the a heterodimer, is while a is enzyme However, synthase 2). ADC bacterial (Fig. the synthase (ADC) chorismate ainpoesi paetyseii opat lhuhbl fpatflts u iohnra hc represent which mitochondria, but folates, plant of bulk elucidated. of form yet storage not a is as serve ester may the although It of plants role to physiological specific the apparently is esterifi- This process 2003). cation al., et (Quinlivan planta in esterified B Branch ABA B yAClae(ihl ta. 99 re tal., et Green 1989; al., et (Nichols ADC by ABA nwiha which forms in polyglutamylated as mainly occur folates Natural derivatives. oteguaaemit.Nmee rohasmr potential mark arrowheads Numbered moiety. glutamate the to si atra h is tpi h ovrino choris- of conversion the in step first the bacteria, in As ie fato fctblcezms(e et.Termvlo the of removal The text). (see enzymes catabolic of action of sites ntetx)i o hw nti figure. this on mentioned shown 4, not is (reaction text) glutamate the first in the after tail polyglutamyl p B smdae npat yaminodeoxy- by plants in mediated is ABA ␥ lne hi fu oaotegtrsde sadded is residues eight about to up of chain -linked Ͻ .1po min pmol 0.01 p B,admyb h ocnrto,weespatd aetelws (Gam- lowest the have plastids whereas concentration, the be may and ABA, Ϫ 1 mg RPSINE O.4,MRHARL2005 MARCH–APRIL 45, VOL. SCIENCE, CROP Ϫ p 1 B a eto ossigo h yoo,nces n aul (Gam- vacuole and nucleus, cytosol, the of consisting tion be may ABA rti) This protein). p B sfrisac npalae,flt sdsrbtdi 30/ a in distributed is folate leaves, pea in instance for is ABA i.2 h erhdooaeboytei aha npat.Teen- The plants. in pathway biosynthesis tetrahydrofolate The 2. Fig. a r oaie nmtcodi Nuugre al., et (Neuburger mitochondria in Re 1996; localized are way Moieties Glutamate omi which in form ytei fFltsfo Pterin, from Folates of Synthesis 2003). al., et (Quinlivan lnsb iyrflt ytaeadsbeun glu- subsequent and in synthase catalyzed dihydrofolate is (Prabhu a step by glutamylation dihydropteroate first plants The product, 1997). own al., et in- its feedback by is synthase hibited dihydropteroate Plant pyrophospho- the bifunc- kinase. in a enzyme hydroxymethyldihydropterin next of the part pathway, contains is also that plants protein in tional which synthase, pteroate h trnand pterin The ayain yflloyltmt ytae(Ravanel synthase folylpolyglutamate by tamylations /7rtoaogmtcodi,clrpat,adafrac- a and compartments; chloroplasts, mitochondria, cell among plant ratio 3/67 all not if most in chloro- present and are mitochondria, cytosol, among split is way Transport and Compartmentation activity. synthase is thymidylate THF has that to reductase also dihydrofolate dihydrofolate bifunctional of a by reduction mediated The 2001). al., et nyafwpreto h elvlm,hv h highest the have volume, cell the of percent few a only the contains fraction latter the Thus, 2001). al., et bonnet aesnhts;HP,hdoyehliyrpei pyrophos- hydroxymethyldihydropterin HPPK, synthetase; folylpolygluta- FPGS, mate synthase; dihydropteroate DHPS, dihydroneopterin aldolase; DHNA, synthase; dihydrofolate DHFS, ductase; underlined. are plants from DHNTP, cloned dihydroneopterin; been DHN, aminodeoxychorismate; yet ADC, not have that zymes fFG swl stemtcodilezm hw (Ravanel shown enzyme mitochondrial isoforms the plastidial 2001). as al., and well et cytosolic as are FPGS there of that Note phokinase. h atfv tp ftepatflt ytei path- synthesis folate plant the of steps five last The iyrnotrntihsht;Gu ltmt;GP guanosine GTP, glutamate; 5 Glu, triphosphate; dihydroneopterin P,hdoyehliyrpei pyrophosphate, hydroxymethyldihydropterin PPi, ys;GPH,GPccoyrls ;DF,dhdooaere- dihydrofolate DHFR, I; cyclohydrolase GTP ADCS, GTPCHI, aminodeoxychorismate phosphate; lyase; ADCL, inorganic synthase; Pi, aminodeoxychorismate pyrophosphate; PPi, benzoate; ؅ tihsht;HDP yrxmtydhdotrn HMDHP- hydroxymethyldihydropterin; HMDHP, -triphosphate; sntwrh httepatflt isnhsspath- biosynthesis folate plant the that noteworthy is ´ beille ´ ta. 97 aae ta. 01 Fg 2). (Fig. 2001) al., et Ravanel 1997; al., et p p B oeisaeculdb dihydro- by coupled are moieties ABA B stafce ihnpatcells plant within trafficked is ABA p B,and ABA, p ABA, p -amino- Reproduced from Crop Science. Published by Crop Science Society of America. All copyrights reserved. motit lsis n ( and plastids; into import ra ( dria; t eea rnpr rcse (e.g., processes medi- transport may several type one ate since transporters of types different all steps, to eight corresponding transporters ( presumably cell are of There the import outside from for ( uptake evidence mitochondria is into there 5-formyl-THF addition, In vacuoles. p hsrqiigtasotmcaim Fg ) These ( 3). following: (Fig. the include mechanisms steps transport transport requiring thus eitsms hrfr oei n u forganelles, of out and in move therefore must mediates nomtcodi;( mitochondria; into indicate Letters plants. in precursors and folates of Transport 3. Fig. stafce ntefr fishdohlcguoeester glucose 2003). hydrophilic al., its of et form (Quinlivan the if in needed a be trafficked may is As transporter a plants. but in membranes, no identified but acid, been 2000), weak hydrophobic Moran, yet and have Titus transporters 1999; Sirot- Tolner, 1999; and al., et nak (Kundig animals from cloned and ized onte l,20) oae n hi isnhtcinter- biosynthetic their and Folates 2001). al., et bonnet c,adpa uigpshretsoae(ct et (Scott storage postharvest during peas spin- and strawberries, for ach, reported been substan- have and de- decreases 2002), tial a al., et shown (Basset have ripening during folate crease fruit development. tomato during of content Measurements folate in changes about or 00 Stra 2000; hrti otavs eln sezmtco due or enzymatic is decline postharvest this ther h hmclbekono oae,adla oae a ujc ofebc niiin(codne l,1992), al., et (Schoedon inhibition feedback to subject con this In 2002). light Gregory, by and promoted cleavage (Hanson oxidative may to folates exposed leaf be and also folates, of breakdown chemical promote the can plastids reac and mitochondria the by principle, produced cies In 2000). depleted al., are et antioxidants (Scott endogenous as breakdown cal B (or ABA nerdtasotses(e et.Abeitosaea nthe in as are Abbreviations text). (see steps transport inferred oaeadpei rnpreshv encharacter- been have transporters pterin and Folate 2. Fig. of legend eyltl skonaotflt unvri plants in turnover folate about known is little Very d oaeepr rmmtcodi;( mitochondria; from export folate ) ˚ lsjo p a B lcs se)ipr nomitochon- into import ester) glucose ABA ¨ - h ta. 03.Hwvr ti nla whe- unclear is it However, 2003). al., et lhuhteeaentncsaiyeight necessarily not are there although , oaeCatabolism Folate b ) p p B xotfo lsis ( plastids; from export ABA B a ifs reyacross freely diffuse may ABA ASTE L:FLT EAOIMADBOOTFCTO NPLANTS IN BIOFORTIFICATION AND METABOLISM FOLATE AL.: ET BASSET f osbyflt motinto import folate possibly ) h Pah ta. 1998). al., et (Prabhu ) g swl sfolate as well as ) a , a ieoye spe- oxygen tive d eto,i is it nection, trnimport pterin ) and , ocei uhan Such chemi- to e g folate ) ). p ABA l,Ove al., in- c ) a ensont rmt ecin1i nmlcells animal in 1 Reaction promote to shown been has iebohmsr n eois o ntne ferritin instance, For genomics. and biochemistry tive enlctdi iohnra(oee l,2002). al., et (Roje has mitochondria and plants in in 2003). located cloned been al., recently been et has mamma- enzyme that (Anguera of This found cycloligase activity been catalytic 5-formyl-THF also secondary lian has a It is pro- plants. 1 this Reaction in ferritin, contain occur plastids could As cess 2000). al., et (Suh ie n le o hi xsec oefo compara- from come existence their for clues and tive, ta. 2001). al., et FPGH ainta lnshv ee htecd ooosof homologs encode that obser- genes the have by plants implied is that 4 vation Reaction 1967). al., (Ko- lumaz- folates et of deaminated bayashi in presence from the originate reactions may by these which implied ines, of is two 2 of Reaction plants. presence the indirect is for There glutamate. evidence first the just leaving tail, myl or ring; the pteridine of the hydrolysis of (3) deamination (2) and cleavage); plants pterin dative the in between scission pathways the enzymatic degradative (1) 1): four (Fig. drawn potentially there Thus, be 1984). are may (Scott, organisms inferences other of some studies from but plants, in 2002). tabolism al., et (Stakhov (6-car- leaves folates pea of in product boxypterin) the photolysis promote a can of irradiation accumulation UVC that note to teresting fctblcezms rb eotn oaesynthesis folate rerouting by or enzymes, catabolic downregulation of by either breakdown, enzymatic chemi- and into decreasing compartment cal (iii) and inert folates stockpiled; be metabolically can of they the other where transport or through the vacuole flux the overall increasing limit (ii) that pathway; transporters) steps by (or mediate synthesis enzymes that more increasing or (i) one overexpressing folate: higher engineering Hague l,19) n uat fan of soybean mutants in and FPGH extracellular 1996), the be al., to et However, found (Huangpu activity. been FPGH has been protein have yet has to homologs folylpoly- these shown of of None (FPGH). types drolase mammalian both oad5fry-H,wihi h otsal natural stable most the is which 5-formyl-THF, toward vrxrsino oaeSnhssEnzymes Synthesis Folate synergistic. of be Overexpression will combination a that and exclusive, likely mutually is not it are strategies These folate. ahasi tnadgmi nmtblcengineering. metabolic in gambit standard a is pathways bacterium sn TCIrlae rmfebc euainis regulation feedback is from released GTPCHI GTPCHI bacterial a As using 2003). al., three- et by (Sybesma production folate fold increased GTPCHI own its oetal oeefcie eaecretyexploring currently are We effective. more potentially eaoi niern fFlt eesi Plants in Levels Folate of Engineering Metabolic hr r orprso nyemdae oaeca- folate enzyme-mediated of reports no are There te erdto ehnssi lnsaespecula- are plants in mechanisms degradation Other ocpuly hr r he oeta taeisfor strategies potential three are there Conceptually, p p B-ltmt;ad()rmvlo h polygluta- the of removal (4) and ABA-glutamate; B-ltmlmite a loocr uigoxi- during occurs also (as moieties ABA-glutamyl epesn h omtigezmsa h edof head the at enzymes committing the rexpressing eeaemrhlgclyanra (Helliwell abnormal morphologically are gene prahhsbe fetv ntelci acid lactic the in effective been has approach atccu lactis Lactococcus p B-ltmllnaeo folates of linkage ABA-glutamyl hrb vrxrsinof overexpression whereby ␥ guaaehy- -glutamate Arabidopsis 451 Reproduced from Crop Science. Published by Crop Science Society of America. All copyrights reserved. fteepeusr nomtcodi.Snentigis nothing Since mitochondria. transport into the precursors by these limited of be still might production folate Compartmentation and Transport Folate Modified dihy- increased the accommodate production. to drofolate low too endogenous is the activity if to reductase necessary be dihydrofolate also may overexpress it 2000), al., et highly (Quinlivan is dihydro- labile dihydrofolate Because minimizing accumulation. thereby pteroate enzyme, synthase, next the overexpressing dihydrofolate by with dealt better therefore be may enzyme engi- this be of reducing inhibition without can Product activity. inhibition synthase product the decrease that to neered unlikely and pterin is the it both substrates, com- of dihydropteroate binding Because blocks 2000). petitively al., mitochon- et in (Scott accumulation dria its preventing and synthesis huhi ed tlati a ev sasoaepool storage a as al- serve unclear, may is it least plants at in seeds role in its though and (Stover 1993) units one-carbon Schirch, of and donor a not is 5-formyl-THF folates, other reaction Unlike hydroxymethyltransferase. side 2000). serine a al., of via et 5,10-methenyl-THF from (Scott formed target is engineering It attractive an is it Pool Folate the of Stabilization organelle. this synthesis into its import of folate inhibition directing feedback by accumu- without to possible are folate be enzymes may late it biosynthetic vacuole, the folate in the present not As vacuole. such the compartment strat- inert as potential an within a folate the also stockpile to flux, is egy transport biosynthetic folate increase of to engineering biosynthesis. potential folate its plant priority Besides of a engineering is transporters metabolic for these cloning and identifying plant about known own its regulating synthase, dihydropteroate of inhibitor by retargeted matched is not is that it synthase cytosol. if the ADC lost to an be would overexpress transport to could any or freeing mechanisms be for thesis regulatory that strategy from regula- synthase one possible ADC potential Thus, is plant plastids. but in 2004), It exist al., 1990). may et tors either Muench, (Basset not mammalian by vitro and does in inhibited in (Matherly lates directly synthase folates be ADC most to plant proteins plants, appear The of in production. overexpression folates toward folate the pyrophos- of channel could be triphosphate aldolase state DHN might and DHN phatase folates overexpressing plant that possi- is ble it Consequently, pathway. stability. synthesis folate points its the branch in favor potential DHN are to 1985), to itself (Brown, DHN expected is pterins and Similar of triphosphate triphosphate number plants. a DHN in to which in precursor folate a in of deregulated bacteria, accumulation be and or enzymes that synthesis mammals to animal cycloligase) of 5-formyl- the (5-formyl-THF expected Enhancement do protein, increase enzyme this plants are to the of as of strategy homologs and activity have One 1998), not 1975). Hatakeyama, al., and protein et regulatory (Yoneyama feedback (Shin feed- special a are by , back-regulated GTPCHIs another of Vertebrate (animals production ). the GTPCHIs to- for vertebrate transgenic GTPCHI have of expressing construction lines the mato with strategy a such 452 vnif Even smnindaoe iyrpeot saproduct a is dihydropteroate above, mentioned As ic -omlTFi h otsal aua folate, natural stable most the is 5-formyl-THF Since p B n trnoepouto r achieved, are overproduction pterin and ABA p B,pei,o oaecarriers, folate or pterin, ABA, RPSINE O.4,MRHARL2005 MARCH–APRIL 45, VOL. SCIENCE, CROP p B rfo- or ABA p ABA tn o ln oae r aaoie n o they how and catabolized stabilized. are be folates can plant under- how to important stand therefore folatesis It overproduced synthesis degraded. if quickly engineering out are wiped by be or- might made between transport gains or relations any whether sink Lastly, and are so, gans. source folates if folate whether and are learn organs, there to plant particularly crucial between also transported is is compartments It precur- needed. subcellular their and among folates of sors in transport investigations. the folates further into of for Research frontiers storage open and remain transport plants hand, other the available. On becoming now are trials pioneering the Results these from synthase. targeted ADC and naturally GTPCHI of have overexpression attempts engineering first tp fteptwyspei and pterin committing pathway’s the the in of reside steps may world. control the substantial around laboratories Since engineering various in metabolic way of under trials are first the and elucidated, proteins. folate-binding in increase syn- an engineering by achieved production folate in gain stability their increasing thus protein-bound, are tissues binding the about known is little While folates. bind that be would also ascorbate in high are that fruits certain 2000). al., et (Scott 5,10-methenyl-THF to it the reconverts suppressing by recycling its block to is content THF eSiiu . .Vna,adAP a on 95 nyi charac- Enzymic 1995. Loon. van A.P. and one-carbon Vankan, and P. A., folate Saizieu, De of world fascinating The 2000. E.A. Cossins, 115–154. p. pterins. of Biosynthesis 1985. G.M. Brown, micronutrient fighting for tool new A breeding: Plant 2002. H.E. Bouis, M. Garza, La De Diaz R. Ziemak, M.J. Quinlivan, E.P. G., Basset, Re F. Ravanel, S. Quinlivan, E.P. G.J.C., Basset, and Selhub, J. Nasrallah, I.M. Ghandour, H. Suh, J.R. M.C., Anguera, h ytei aha fflt npat snwlargely now is plants in folate of pathway synthesis The of stabilization the for strategy potential Another hmcldpopoyaino iyrnotrntriphosphate. 306:371–377. dihydroneopterin J. of Biochem. dephosphorylation chemical a of terization 78:691–708. Bot. J. Can. & metabolism. Wiley John 2. Vol. Pterins York. and New Folates Sons, (ed.) Benkovic S.J. and 132:491S–494S. Nutr. J. Proc. malnutrition. I. cyclohydrolase 99:12489–12494. GTP USA bimodular branch Sci. pterin unique Acad. the a Natl. of step by first mediated The is plants: Hanson. in A.D. synthesis and Folate Gregory, 2002. USA. J.F. Bacher, Sci. A. Acad. Schiffmann, S. Natl. 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