Communi cations

Biomass Conversion DOI : 10.1002/anie.20100468

Mechanism of !ucose Isomeri"ation #sin$ a %o!i& 'e(is )ci& Cata!*st in +ater,,

9

-uri* .omn/'esh0ov1 Manue! Mo!iner1 2a* ). 'a3in$er1 an& Mar0 4. Davis if .ecent!*1 ( e ,5167h e conversion a!&eh*&e to create o
the re!ate& a/ re8orte& on a tin/ of $!ucose into h*&ro=* 0etone containin$ "eo!ite 9tin fructose for the 8ro&uction of hi$h/ 9%cheme 1<.54 67his in the frame(or0 of a 8ure/si!ica ana!o$ of fructose corn s*ru8s mechanism can occur either 3* a "eo!ite 3eta1 &enote& 9:;C%< is the E %n/Beta< that is a !ar$est 3iocata!*tic 8roton transfer 5 8rocess in the (or!&1 9%cheme 1 ) < o r 3 hi$h!* active materia! D r. an& it recent!* has * a n intramo!ecu!ar for the isomeri"ation5E6 5,6 Dr. -. .omn/'esh0ov1 2 3een consi&ere& as h*&ri&e shift . a 0e* interme&iate 9%cheme 1 B<. ) %evera! stu&ies ste8 in the . have sho(n that conversion of ' 3asecata!*"e& 3iomass to fue!s an& a isomeri"ations ta0e chemica!s.51a67his 3i 8!ace 3* a 8roton reaction is t*8ica!!* n transfer mechanism $ cata!*"e& 3* an throu$h a series of e immo3i!i"e& eno!ate r1 en"*me1 =*!ose F interme&iates isomerase1 that r $enerate& after the $enerates an o &e8rotonation of the e>ui!i3rium mi=ture f. a/car3on*! car3on in of 42 (t ? fructose1 M @ 0 ( t ? $!ucose1 (ater.5@67h e =*!ose . 4 an& 8 ( t ? other isomerase/cata!*"e& 8rocess is a!so . sacchari&es. 7he D en"*matic 8rocess thou$ht to 3e a is hi$h!* se!ective1 me&iate& 3* eno!ate vi interme&iates 3ut it has severa! s $enerate& 3* &ra(3ac0s that D histi&ine/&irecte& increase 8rocessin$ iv costs1 inc!u&in$ the 3ase cata!*sis C561 is D6ho(ever1 recent io use of 3ufferin$ n so!utions to maintain stu&ies have sho(n that meta! centers in o 8:1 narro( f the en"*me are o8eratin$ C tem8eratures1 strict res8onsi3!e for the h sta3i!i"ation of the fee& 8urification e su$ars o8en/chain re>uirements1 an& m form an& the 8erio&ic is tr re8!acement of the su3se>uent a!&oseB 0etose isomeri"ation * en"*me &ue to 3* (a* of an a irreversi3!e n intramo!ecu!ar &eactivation. & h*&ri&e shift. 526Inor$anic 3ases can a!so cata!*"e this reaction1 a!3eit (ith !o( *ie!&s &ue to the re&uce& sta3i!it* of monosacchari&es in the 8resence of 3asic cata!*sts.5A67h e mechanism of this a!&oseB0etose isomeri"ation invo!ves h*&ro$en transfer from C/2 to C/1 an& from O/2 to O/1 of an a/h*&ro=*
C h e m ic a! 4 n $i n e e ri n $ C a! r ni a I n st it u t e o f 7 e c h n o! o $ * F a s a & e n a 1 C ) 9 1 1 2 @ 9 # % ) < ; a = : 9 E 1 < 6 2 6 / @ 6 8 / 8 D 4 A 4 / m ai

!: m & a vi s G c h e m e .c a! t e c h . e & u 6 7hese authors contri3ute& e>ua!!* to this (or0. 5,,6 7his (or0 (as financia!!* su88orte& as 8art of the Cata!*sis Center for 4ner$* Innovation1 an 4ner$* ;rontier .esearch Center fun&e& 3* the #.%. De8artment of 4ner$*1 Office of %cience1 Office of Basic 4ner$* %ciences un&er )(ar& Hum3er D4/ %C00010004. M.M. ac0no(!e&$es ;un&acin .amn )reces Fost&octora! .esearch ;e!!o(shi8 Fro$ram for financia! su88ort.

%u88ortin$ information for this artic!e is avai!a3!e on the +++ un&er htt8 ://&=.&oi.or$/10.10 02/anie.20100468 9.17his cata!*st

(as sho(n to 3e active over a (i&e tem8erature ran$e 9A4AB 41A I<1 in aci&ic so!utions1 an& (ith $!ucose fee&s as hi$h as 4@ (t ? t o $ive

8ro&uct *ie!&s e>uiva!ent to those o3taine& (ith =*!ose isomerase 9at its 8referre& reaction con&itions<. :ere1 usin$: an&1A596C HM. s8ectrosco8* on isoto8ica!!* !a3e!e& $!ucose1 ( e &emonstrate that in the 8resence of %n/ Beta1 the isomeri"ation

reaction in (ater 8rocee&s 3* (a* of an intramo!ecu!ar h*&ri&e shift. )!thou$h 'e(is aci&it* is usua!!* su88resse& 3* the 8resence of (ater1 verification of this mechanistic 8ath(a* confirms that frame(or0 tin centers in %n/ Beta act as 'e(is aci&s in

a>ueous me&ia. Frevious re8orts have sho(n stron$ interactions 3et(een 'e(is aci& centers in "eo!ites an& h*&ro=*/car3on*! moieties in reactants &isso!ve& in or$anic so!vents. 5106;o r e=am8!e1 Corma et a!. use& %n/Beta to cata!*"e the Meer(einB

Fonn&orfBJer!e* 9MFJ< re&uction 89@4 2010 +i!e*/ JC: of car3on*! Jer!a$ m3: K Co. I a)1 com8oun&s in +einhei m )n$e(. Chem . methano!15116an& Int. 4&. 2010 1 491 89@4 B 89@D5E6Dr. M . 7aarnin$ et a!. Mo!iner1%cheme 1. use& %n/Beta for !ucose isomeri"ation the 8ro&uction ofmechanisms 3* (a* of )< !actate 8roton transfer an& B< &erivatives from intramo!ecu!ar h*&ri&e monosacchari&e shift. s in methano!.7h 58 6of $!ucose into e MFJ reaction fructose in a>ueous me&ia mechanism invo!ves the simu!taneous coor&ination

1A

11AC

an&

Angewandte

1A

CB
1A

C B5106of the car3on*! $rou8 of a 0etone an& the 51A6Christensen et a!. recent!* re8orte& on the h*&ro=* $rou8 of an a!coho! to a meta! center. isomeri"ation of &ih*&ro=*acetone usin$ %n/Beta in )&e>uate 'e(is aci&it* in the meta! center a!!o(s methano! an& in (ater1 (here it (as o3serve& that no the 8o!ari"ation of the car3on*! $rou8 of the 0etone !oss in activit* occurre& in methano!1 3ut severe an& 8romotes a h*&ri&e shift from the h*&ro=* cata!*st &eactivation occurre& in (ater &ue to the $rou8 of the a!coho! to the car3on*! $rou8 of the formation of !ar$e car3onaceous &e8osits. 7he 0etone 9see ;i$ure %1 in the %u88ortin$ mechanism of reaction in (ater (as not Information<.5126Corma an& co(or0ers have sho(n e=8!aine&.7hus1 the (or0 re8orte& herein is the first to that meta! centers 9H31 7a1 an& %n< incor8orate& &escri3e the mechanism (here3* %n/Beta acts as a into "eo!ites can act as 'e(is aci&s (hen use& in 'e(is aci& in a 8ure!* a>ueous environment.Chemie or$anic so!vents containin$ a sma!! amount of (ater. : HM. s8ectrosco8* (ere use&

to stu&* the isomeri"ation of $!ucose/D2 (ater at A8A I for 1@ min in the 8resence of usin$ %n/Beta as the cata!*st. ) com8arison %n/Beta1 the $!ucose an& fructose fractions ofC HM. s8ectra of un!a3e!e& $!ucose an& (ere se8arate& usin$ hi$h8erformance !i>ui& $!ucose/D2 fee& so!utions revea!s that chromato$ra8h* 9:F'C<. )na!*sis of resonances o3serve& at & L D4.1 an& D1.A C HM. s8ectrum for each fraction sho(s 88m in the un!a3e!e& $!ucose so!ution a88ear that after reaction1 $!ucose/D2 remains as !o(/intensit* 1 : 1 : 1 tri8!ets in the unchan$e& 9;i$ure 1 31c<1 (hi!e the fructose $!ucose/D2 so!ution 9;i$ure 1 a13<. 7his effect 8ro&uct has si$nificant &ifferences com8are& is re!ate& to the &isru8tion of the nuc!ear to an un!a3e!e& fructose stan&ar& 9;i$ure 1 Overhauser enhancement 9HO4< 3* the e1f<. %8ecifica!!*1 resonances at & L 6A.8 an& &euterium atoms in the C/2 8ositions of the 62.6 88m assi$ne& to the C/1 8osition of the t(o confi$urations of $!ucose/D2 in so!ution 3 /8*ranose an& 3/furanose confi$urations in 93 /8*ranose an& a /8*ranose1 8resent in a 6 the un!a3e!e& fructose stan&ar& a88ear as 4 :A6 ratio<. Durin$ HO41C resonance !o(/intensit* tri8!ets for the fructose 8ro&uct intensities are enhance& u8 to 200 ? for recovere& after reaction. &irect!* 3on&e& : HM. s8ectra for 3oth su$ars further confirm these : 3roa&/3an& resu!ts. 7h e: HM. s8ectra of $!ucose/D2 3efore &ecou8!in$ is use& to su88ress C1 : an& after reaction remain constant 9;i$ure 2 31c<1 cou8!in$s Cho(ever1 this resonance (hi!e the fructose s8ectrum sho(s the am8!ification is not o3serve& for: 8airs an& &isa88earance of the resonance at & L A.4@ 88m resonances associate& (ith C/2 in $!ucose/ &ue to the 8resence of a &euterium atom in the C/1 D2 are thus su3stantia!!* &iminishe&. )fter 8ositio reactin$ a 1 0 ( t ? so!ution of $!ucose/D2 in 1A n !ucose &euterate& at 1A $!ucose/D2 C HM. s8ectra heate& of in (ater at 9A8A I< reaction in the a3sence tem8erature of at cata!*st isoto8ic revea! scram3!in$ that no occurs 9;i$ure im8ortant %2 3ecause B<. 7his it resu!t is &emonstrates C D 3on& the in inertness (ater reaction in&icatesthat con&itions an* isoto8ic an& rearran$ement mo!ecu!e &urin$ (ithin the reaction the is of entire!* the cata!*st. &ue to 1 : the 8airs actions (hen the C/2 8osition 9$!ucose/ D21 ;i$ure %2 )< (as use& to 8erform HM. stu&ies on the isomeri"ation reaction. )s sho(n in %cheme 11 the 8roton at the C/2 8osition 1A1 8!a*s a fun&amenta! ro!e 5146 in the reaction mechanism re$ar&!ess of (hich 8ath(a* is fo!!o(e&. In&ee&1 isoto8ic su3stitution at the C/2 8osition resu!te& in a t(o/ fo!& &ecrease in the initia! reaction rate 90:/ 0DL 1.981 ;i$ure %A<1 revea!in$ a consi&era3!e 0inetic isoto8ic effect. Im8ortant!*1
21A

;i$ure 1.C HM. s8ectra of a< un!a3e!e& $!ucose1 3< !a3e!e& $!ucose/D2 (ith %n/Beta1 an& $< fructose fraction o3taine& $!ucose/D21 c< $!ucose fraction o3taine& after reactin$ after reactin$ !a3e!e& $!ucose/D2 (ith HaO:. $!ucose/D2 (ith %n/Beta1 &< $!ucose fraction o3taine& after89@@)n$e(. Chem. Int . 4&. 2010 1 49 1 89@4 B89@ D 2010 reactin$ !a3e!e& $!ucose/D2 (ith HaO:1 e< un!a3e!e& +i!e*/JC: Jer!a$ m3: K Co. I a)1 +einheim fructose1 f < fructose fraction o3taine& after reactin$ !a3e!e&(((.an$e(an&te.or

Communi cations

;i$ure 2.1

C a n &

1111951@6%n/HM.<

1A

1 A

that 9;i$ure %4 B<. In&ee&1 iso!ate& frame(or0 tin %nO22is 0no(n to 3e centers in "eo!ites are hi$h!* sta3!e in aci&ic an& res8onsi3!e for &rastica!!* 3asic me&ia an& 8ro3a3!* enhancin$ the rates of &oes not 8artici8ate in the certain reactions.;o r "eo!ite cr*sta!!i"ation $!ucose isomeri"ation in 8rocess 9there3* (ater1 (e o3serve& that 8rovi&in$ e=traframe(or0 frame(or0 tin centers are %nOnano8artic!es (ithin necessar* for %n/Beta to the "eo!ite 8ores<. It has cata!*"e the reaction. %n/ 3een su$$este& that the Beta s*nthesi"e& 3* a actua! active site in %n/ 8roce&ure 0no(n to fu!!* Beta is a 8artia!!* incor8orate tin into the h*&ro!*"e& frame(or0 tin frame(or0 9%nC!4as the s8ecies 9i.e. a tin source< is hi$h!* active 9%iO<A25166%n9O:< center<. for the isomeri"ation 5D6Interestin$!*1 reaction1 (hereas %n/ Iova!evs0* et a!. sho( Beta s*nthesi"e& usin$ that ana!o$ous %nOas the tin source is h*&ro!*"e& meta! s8ecies com8!ete!* inactive in =*!ose isomerase 8!a* 9;i$ure %4 )<. 7h e a fun&amenta! ro!e in &iffuse ref!ectance #J/Jis 8rotonationB s8ectrum for the active &e8rotonation se>uences materia! sho(s a sin$!e (ith (ater an& $!ucose 3an& centere& at 220 nm1 mo!ecu!es &urin$ the assi$ne& to tetrahe&ra!!* isomeri"ation coor&inate& meta!1 (hi!e 8rocess.I&entif*in$ the s8ectrum for the non/ simi!arities 3et(een the active materia! sho(s a active sites in the 3an& centere& at A00 nm1 3io!o$ica! an& the %n/ assi$ne& to octahe&ra!!* Beta cata!*st s*stems is coor&inate& meta! in one of our current e=tra/frame(or0 8ositions o3Mectives

.: HM. s8ectra of a< un!a3e!e& $!ucose1 3< !a3e!e& $!ucose/D21 c< $!ucose fraction o3taine& after reactin$ $!ucose/
D2 (ith %n/Beta1 &< $!ucose fraction o3taine& after reactin$ !a3e!e& $!ucose/D2 (ith HaO:1 e< un!a3e!e& fructose1 f < fructose fraction o3taine& after reactin$ !a3e!e& $!ucose/D2 (ith %n/Beta1 an& $< fructose fraction o3taine& after reactin$ !a3e!e& $!ucose/D2 (ith HaO:.

9;i$ure 2 f<. Inte$ration of the areas in these s8ectraconfirms the 8resence of si= CB: 8airs for the $!ucose

an& fructose fractions in contrast to the seven CB: 7his stu&* 8roves that %n/Beta can act as a 'e(is 8airs foun& in un!a3e!e& $!ucose or fructose. 7hese aci& ca8a3!e of cata!*"in$ the isomeri"ation of resu!ts c!ear!* in&icate that the &euterium atom $!ucose in a 8urea>ueous me&ium. ;urther stu&ies !ocate& in the C/2 8osition of $!ucose/D2 has move&are necessar* in or&er to un&erstan& the nature of to the C/1 8osition of fructose1 there3* une>uivoca!!* the interactions amon$st the active site in %n/Beta1 &emonstratin$ that the $!ucose isomeri"ation reaction the su$ar an& the so!vent &urin$ the isomeri"ation (ith a so!i& 'e(is aci& cata!*st in 8ure (ater 8rocee&s reaction. Frevious re8orts 3* Corma an& co/ 3* means of an intramo!ecu!ar h*&ri&e shift. (or0ers have sho(n 9usin ) simi!ar s8ectrosco8ic stu&* (as 8erforme& usin$ so&ium h*&ro=i&e 9HaO:< as a 3asic cata!*st. ) 1 0 ( t ? a>ueous so!ution of $!ucose/D2 (as reacte& in the 8resence of HaO: 90.1 m < a t A8A I for 2 min.1: HM. s8ectra of the $!ucose an& fructose fractions sho( consi&era3!e &ifferences (hen com8are& to the resu!ts o3taine& (ith %n/Beta. ;irst1 the: HM. s8ectra for the un!a3e!e& fructose an& for the fructose fraction iso!ate& after reaction sho( no &ifferences1 in&icatin$ that the fructose fraction &oes not contain &euterium atoms 9;i$ure 1 e1$ an& ;i$ure 2 e1$<. %econ&1 theC HM. s8ectrum of the $!ucose fraction sho(s $!ucose/D2 mi=e& (ith a sma!! amount of re$u!ar $!ucose1 a s in&icate& 3* the 8resence of a sma!! resonance at & L D4.1 88m 9see inset in ;i$ure 1 &<. 7h e 8resence of un!a3e!e& $!ucose is corro3orate& 3* the a88earance of a resonance at & L A.1 88m in the: HM. s8ectrum1 (hich is assi$ne& to a 8roton in the C/2 8osition 9;i$ure 2 &<. 7hese resu!ts c!ear!* in&icate that the 3asic cata!*st o8erates 3* a 8roton/transfer mechanism (here3* the &euterium atom is remove& from the a/car3on*! car3on of $!ucose/D2 to form the corres8on&in$ eno!ate1 an& a 8roton from so!ution is su3se>uent!* re/incor8orate& into the mo!ecu!e1 *ie!&in$ some un!a3e!e& $!ucose a!on$ (ith the un!a3e!e& fructose 9%cheme 1 )<. $89@6 (((.an$e(an&te.or$ 201 0 +i!e*/JC: Jer!a$ m3: K Co. I a)1 +einheim )n$e(. Chem . Int. 4&. 2010 1 491
89@4 B89@D

4=8erimenta! %ection
51@6

.a88o8ort<1 +i!e*1 Chichester1 19901 8 8. A99 B

%n/Beta (as s*nthesi"e& fo!!o(in$ 8roce&ures 480. Chemie 8revious!* re8orte& in the !iterature 9see %u88ortin$ Information for &etai!s<.Isomeri"ation e=8eriments (ere carrie& out in 10 m' thic0/(a!!e& $!ass reactors 9J+.< heate& in a tem8erature/contro!!e& oi! 3ath. 1.@ $ o f a 1 0 5@6 a< I. ). .ose1 4 . ' . OConne!!1 . . F. Mort!oc01 Biochim. Bio8h*s. )cta 4n"*mo!. 19691 1D81 AD6 C ( t ? $!ucose/D2 9Cam3ri&$e Isoto8es Cor8.< so!ution an& the corres8on&in$ cata!*st amount to achieve a 1 :@0 3 < I . 2. %chra*1 I . ) . .ose1 Biochemistr* 19D11 101 10@8. meta! :$!ucose mo!ar ratio (ere a&&e& to the reactor1 566 a< I. H. )!!en1 ). 'avie1 . ;ar3er1 ). !asfe!&1 sea!e&1 an& heate&. ;urther information a3out the cata!*tic tests an& characteri"ation techni>ues can 3e foun& in the . Fets0o1 D. .in$e1 Biochemistr* 19941 AA1 1481. 5D6 ). -. Iova!evs0*1 ' . :anson1 %. O. ;isher1 M. %u88ortin$ Information. Must*a0imov1 %. ). Mason1 J. 7revor ;ors*th1 M . F. B!a0e!e*1 D. ) . Ieen1 7. +a$ner1 :. '. Carre!!1 .eceive&: 2u!* 291 2010 ). I. Iat"1 2. F. !us0er1 F. 'an$an1 %tructure Fu3!ishe& on!ine: Octo3er 20101 181 688. 201 2010 586 M. Mo!iner1 -. .omn/'esh0ov1 M . 4 . Davis1 Froc. Hat!. )ca&. %ci. #%) 20101 10D1 6164. 596 ). Corma1 :. arcia1 Chem. .ev. 20021 1021 A8AD. 5106 a< ). Corma1 M. 4. Domine1 ' . Hemeth1 Ie*(or&s : 3iomass conversion N $!ucose %. Ja!encia1 2. )m. isomeri"ation N HM. s8ectrosco8* N so!i& 'e(is Chem. %oc. 20021 1241 A194 C 3 < M . Boronat1 aci&s N "eo!ites ) . Corma1 M. .en"1 2. Fh*s. Chem. B 20061 1101 21168. 5116 M. %. :o!m1 %. %aravanamuru$an1 4. 7aarnin$1 516 a< %. Bhosa!e1 M . .ao1 J. Desh8an&e1 Micro3io!. .ev. %cience 20101 A281 602. 19961 601 280 C 3 < #%D)1 %u$ar an& %(eeteners : 5126 a< ). Corma1 M. 4. Domine1 %. Ja!encia1 2. Mar0et Out!oo01 i n htt8.// Cata!. 200A1 21@1 294 C 3< ). Corma1 M. .en"1 (((.ers.us&a.$ov/Briefin$/%u$ar/mar0etout!oo0.htm. Chem. Commun. 20041 @@0 C c < ) . Corma1 ;. P. 2010 C c< 2. H. Chhe&a1 . +. :u3er1 2. ) . Dumesic1 '!a3res i Pamena1 C. Fresti8ino1 M . .en"1 %. )n$e(. Chem. 200D1 1191 D298 C )n$e(. Chem. Int. 4&.Ja!encia1 2. Fh*s. Chem. C 20091 11A1 11A06. 200D1 461 D164 C & < -. .omn/'esh0ov1 C. 2. Barret1 O. -.51A6 4. 7aarnin$1 %. %aravanamuru$an1 M. %. :o!m1 'iu1 2. ). Dumesic1 Hature 200D1 44D1 982. 2. Pion$1 .*an M. +est1 C. :. Christensen1 526 a< B. -. -an$1 . . Mont$omer*1 Car3oh*&r. .es. Chem%usChem 20091 2 1 62@. 19961 28012DC 3< . De +it1 ). F. . Iie3oom1 :. van 5146 D. Heuhaus1 M. F. +i!!iamson1 7he Huc!ear Be00um1 Car3oh*&r. .es. 19D91 D41 1@D. Overhauser 4ffect in %tructura! an& Conformationa! 5A6 .. Ha$ors0i1 2. F. .ichar&1 2. )m. Chem. %oc. 20011 )na!*sis1 JC:1 +einheim1 1989. 12A1 D94. 546 2. .. Ieeffe1 ) . 2. Ires$e in 7he Chemistr* 51@6 ). Corma1 M. 4. Domine1 ' . Hemeth1 %. of 4no!s 94&. : O . Ja!encia1 Hature 20011 4121 42A. 5166 M. Boronat1 F. Conce8cin1 ). Corma1 M. .en"1 %. Ja!encia1 2. Cata!. 200@1 2A41 111 .89@D)n$e(. Chem. Int . 4&. 2010 1 49 1 89@4 B89@ D 2010 +i!e*/JC: Jer!a$ m3: K Co. I a)1 +einheim

Angewandte

(((.an$e(an&te.or$