Cc t bo nhin liu sinh hc enzyme bi Plamen Atanassov, Chris Apblett, Scott Banta, Susan Brozik,Scott Calabrese Barton, Michael

Cooney, Bor Yann Liaw, Sanjeev Mukerjee, v Shelley D. Minteer. S nh th no nu bn c th sc pin in thoi di ng ca bn ngay lp tc bng nc ngt ca bn vo n? Cc ng dng nh vy c th l mt con ng tt lu, nhng Vn phng Khng qun M nghin cu khoa hc c u t vo mt tng lai nh by gi. Theo mt sng kin nghin cu i hc nhiu, gio s i hc t khp ni trn c nc ang tp trung vo mt chng trnh nghin cu nm nm xem xt cc thch thc k thut xung quanh mt t bo nhin liu m s chy trn cc loi ng n gin nh nhng ngi tm thy trong thc phm hng ngy ca chng ti.

Hinh 1 .S tng qut ca mt t bo nhin liu sinh hc enzyme bao gm mt cc dng, xc tc bi oxidase thch hp cho vic chuyn i nhin liu ca s la chn hoc mt phc tp ca cc enzym nh vy cho mt qu trnh oxy ha hon ton ca nhin liu sinh hc. Cc m thng c tnh nng oxidoreductase s dng oxy phn t nh cc nhn in t cui cng v xc tc gim nc trong mi trng trung tnh hoc hi chua.

Nhng thch thc l rt ln. Hu ht cc t bo nhin liu th gii hin nay chy bng hydro. Tuy nhin, nh cc nhin liu tr nn phc tp hn, qu trnh oxy ha ny tr nn bao la phc tp hn. . Mt khi nguyn t carbon l trong nhin liu carbon monoxide ca cht xc tc t bo nhin liu c trng tr thnh vn . Cc nh nghin cu ang chuyn sang th gii t nhin trong mt n lc xem cch ng b oxy ha bi cc ng vt sn xut in. S dng cc enzym (cht xc tc t nhin) dng nh l cu tr li, v chng khng b cc vn nhim ca cc cht xc tc kim loi truyn thng . Chng cng v cng phong ph v r sn xut, mt ci g m cc ngnh cng nghip ru vang v cht ty ra c bit n trong nhiu thp k. Nhng p dng cc enzym ng dng in t l rt khc nhau t vic cc enzyme lm sch qun o ca chng ti. i vi mt, cc enzyme khng t b in t d dng nh cht xc tc kim loi lm, m c ngha l to ra mt dng in t cc enzyme l kh khn hn nhiu. Cc enzyme c th c thc hin t b in t ca chung vi trung gian, nhng s dng trung gian c th gy ra cc vn khc trong cc t bo nhin liu. Men cng khng c s dng t. ng vt, enzyme tn ti t do trong cc t bo ca c th, nhng lm vic trong mt t bo nhin liu, chng phi c t trong mt v tr c th v li , mt qu trnh m cc nh khoa hc gi immobilizing cc enzym.cui cng, enzim thng tn ti di lu. Mt enzyme in hnh trong c th con ngi ch ko di mt vi ngy, nhng c hiu qu trong mt my tnh xch tay hoc mt chic t, mt loi enzyme s phi tn ti nhiu thng hoc nhiu nm trc khi cn thay th. Nhng li ch ca cng ngh ny l ln nh nhng ri ro, tuy nhin. Enzyme, nh chng ti cp trc, c gi r v phong ph.chng cn mu xanh l cy, v c th c pht trin v s lng bt c khi no h l cn thit, nh tri ngc vi cc cht xc tc kim loi, cn c khai thc v tinh ch s dng cc quy trnh thn thin mi trng tn km v t. H cng "chn lc", mt t m

cc nh khoa hc s dng m t kh nng ca mt loi enzyme lm vic vi mt nhin liu rt c th, v ch c nhin liu, do , sn phm ph ca mt bc qu trnh oxy ha c th l nhin liu cho cc enzyme khc. Bng cch thc hin bc ny, cc enzyme c th hnh dung c sao chp nhng g ng vt bit lm th no lm: chuyn i ng thnh ch nc v carbon dioxide. Sau khi tt c, c nhiu nng lng trong mt chic bnh rn jelly nh bn c th tm thy trong 77 pin in thoi di ng. Nu bn c th nhn c nng lng ra, n c th c kt qu kh, , ngt ngo. Cc t bo nhin liu sinh hc enzyme c bn cha rt nhiu cc thnh phn tng t nh mt t bo hydro / oxy nhin liu: mt cc dng. mt cc m, v mt du phn cch. Tuy nhin, thay v s dng in xc tc kim loi cc dng v cc m, cht in c s dng l cc enzym oxidoreductase. y l mt lp hc ca cc enzyme c th kch thch phn ng oxy ha kh. V cc enzyme c tnh chn lc Cht in, phn cch c th l mt gii php in phn, gel, hocpolymer. Hnh 1 cho thy mt s ca mt t bo nhin liu sinh hc chung oxy ha glucose nh nhin liu ti bioanode v gim oxy nc ti biocathode. Cc t bo nhin liu sinh hc u tin c gii thiu vo nm 1911 khi Potter men nui cy v cc t bo E. Coli trn in cc bch kim, nhng mi n nm 1962 trong cc t bo nhin liu sinh hc enzyme c pht minh s dng glucose oxidase enzyme oxy ha glucose anode.Trong cui cng 45 nm qua, nhiu ci tin c thc hin trong cc t bo nhin liu sinh hc enzyme v nhng ngi c th c tm thy mt s xt articles.-Tuy nhin, vn cn mt s vn chnh xem xt vi cc t bo nhin liu sinh hc. Chng bao gm thi gian sng ngn hot ng, mt nng lng thp, v hiu qu thp do thng ch kt hp mt enzym lm mt qu trnh oxy ha mt phn nhin liu sinh hc phc tp. C mt s gii php gii quyt nhng vn ny,

nhng nhm ca chng ti l tp trung vo cc gii php cho c nh v n nh ca cc enzym, k thut ca cc enzym hot ng ti u b mt in cc, vn chuyn in t gia enzyme v cc nh su tp hin ti, ti u ha h thng multienzyme, v pht trin cc c tnh tiu chun ha giao thc cho cng ng nghin cu t bo nhin liu sinh hc ni chung. C nh enzyme v n nh Mt phng Php c nh enzym v n nh c s dng cc polyme micellar. Enzyme trong dung dch thng hot ng trong mt vi gi n vi ngy. C th c m rng n 7-20 ngy bng trong lin kt hydrogel v vi in cc surfaces.4 Tuy nhin, cc nh nghin cu ti i hc Saint Louis m rng enzyme hot ng ti b mt in cc ln hn mt nm bng cch c nh trong micellar k nc sa i polymers.7-9 polyme Micellar, chng hn nh, Nafion v chitosan, c th c sa i thch ng k nc l micellar hoc cu trc ti c kch thc ti u cho c nh enzym, trong khi cng m bo mt k nc v m pH vi mi trng cho hot ng ca enzyme ti u. Chin lc ny c chng minh c hai tng tui th hot ng ca enzym, nhng cng lm tng hot ng enzym ca enzyme ln n 2,5-fold.10 Thit k thch hp ca enzyme in cc l mt iu quan trng cho vic to ra cc h thng bioelectrochemical mi nh t bo nhin liu sinh hc v cm bin sinh hc. Cc h thng ny thng lin quan n cc tha thun phc tp polyme ofimmobilization, trung gian oxi ha kh , andenzymes phi tng tc vi b mt in cc. Ti i hc Columbia, cc nh nghin cu ang s dng k thut protein thit k a chc nng protein v peptide c th c hai n gin ha v ci thin in enzym. kt thc ny, h ang to ra cc protein c th t lp rp thnh hydrogel hot tnh sinh hc vi cc hot ng enzyme oxy ha kh. iu ny gip loi b s cn thit cho s kt hp ca cc loi polyme trong h thng. Ngoi ra, cc peptide c th lin kt trung gian oxi ha kh

cng ang c thit k. Cch tip cn k thut phn t ny nn n gin ha ng k vic ch to, c tnh, v lp li ca giao din bioelectrocatalytic s dng trong cc thit b tng lai. Vn chuyn in t. Vn quan trng trong cc t bo nhin liu sinh hc, cng nh trong bt k loi t bo nhin liu lowtemperature, l xc tc ca cc qu trnh in cc. Qu trnh oxy ha ca nhin liu, hy ni ng, xc tc bi cc enzyme nh glucose oxidase (enzyme ca cm bin sinh hc yu thch ca mi thi i) c th c thc hin trc tip hoc c th lin quan n ha gii oxi ha kh. Qu trnh oxy ha enzyme trc tip yu cu cc trang web hot ng ca enzym lin lc ngay lp tc vi b mt in cc. iu dng nh khng th thay cho glucose oxidase bi v n hot ng c "chn vi" qu su trong cc protein "v" ca phn t enzyme kh ln. Phng th nghim quc gia Sandia gii quyt vn ny bng cch sa i v mt di truyn cc enzyme lm a n d tip cn hn tip xc vi cc in cc. Rt nhiu protein t bin c th c thc hin trong mt ngy. Bit c ai s lm vic l mt phn ca vn . Sng lc nhanh cc enzyme bin i gen i vi ti sn electrocatalytic ca h l nhim v ca mt n lc hp tc gia cc phng th nghim quc gia Sandia v i hc New Mexico. Cc nh nghin cu ti i hc New Mexico cng ang xem xt cc vn cc men tiu ha / in cc tng tc t mt gc khc nhau: chng ta hy ri khi enzyme mt mnh, c nhng iu m chng ta c th lm vi cc in cc ca chnh n. Vt liu cu trc nano, c bit l cc ng nano carbon, c th c s dng. t nht l h c th so snh kch thc vi cc enzyme v cc khuyt tt ca h c th l mt phn tt tng tc vi cc trang web hot ng trc tip. Qu trnh oxy ha trc tip ca ng glucose oxidase c nh trong mt carbon ng nano bin i ma trn xp cho thy chng ta c th lm

cho mt anode hot ng ti khong 400 mV (so vi Ag / AgCl) .11 iu ny c ngha rng chng ta c th bt u "t chy" nhin liu ti mt tim nng rt gn vi mt trong nhng cung cp thng tin cc nhit ng lc hc ca h thng (xem hnh. 2). Khi chng ti kt hp vi gim trc tip ca oxy, c xc tc bi c cha ng enzyme (laccases, bilirubin oxidase, hoc ascorbate oxidase) ti mt tim nng ch 50 mV di gi tr nhit ng lc hc, iu ny cho php chng ta ty chn lm cho mt t bo nhin liu sinh hc vi in p nh mch m cao nh 1 V. i vi tt c iu ny xy ra, rt nhiu th cn phi c iu chnh, b mt men tng tc nht ca tt c. Qu trnh chuyn giao v nh hng enzyme l v hn tm quan trng trong chuyn in t trc tip. Phn cn li l "n gin" k thut ca cc ti liu, m nn cho php Hi qun Hoa K v nh nhiu hin nay cng tt

hnh 2. Nguyn tc thit k t bo nhin liu sinh hc ting t co tim nng oxy ha ti a cho ng v tim nng nhit ng lc hc tng ng gim oxy pH trung tnh. Tim nng oxi ha kh ca nhiu enzym v ng yu t tng ng ca h c hin th cng vi cc tim nng "khu vc" c tim nng oxi ha kh ca cc cht trung gian thng thng. ng cong phn cc miu t mn trnh din

hin nay in hnh cho chuyn in t trc tip v trung gian trong cc in cc t bo nhin liu sinh hc. C thc s l mt cch khc: chng ta c th s dng trung gian. y l oxi ha kh cp c th d dng giao tip vi c hai enzym v cc in cc. Vic s dng cc cht trung gian nn c o bi v nu tim nng ca chng l qu gn ca cc enzyme, ng lc ca cc enzyme / trung gian ha gii "tng" s l qu thp. Nu tim nng ca chng ri qu xa ca v tr hot ng enzyme, in p ca pin s b nh hng (xem hnh. 2). Mt nhm i hc bang Michigan ang nghin cu nhng tc ng ca sng lc cc cht trung gian oxi ha kh (da trn hp Osmium) c th h tr c hai qu trnh anode v cathode. Th oxi ha kh nhng cht trung gian l du dng bng cch thay i nh trong ha cht cu trc ca cc phi t hu c. Tt c u c trin khai nh chui bn ca mt polymer oxi ha kh hnh thnh mt khng hydrogel b mt in cc. Qu trnh vn chuyn trong nhng cht hydrogel v trong cc phng tin truyn thng tn xp ca enzyme in cc t bo nhin liu sinh hc (ngh v n nh mt ming bt bin) l mt ch ring ca mnh. S hiu bit su sc v tc ng giao thng trn ng hc s cung cp cho cc nhm phng tin gii quyt nhng vn k thut "n gin" v sn xut nh hin nay nhiu cng tt t cc in cc bioenzymatic. Cc con s hin nay ang chuyn t A mA/cm2. Oxy ha hon ton ca nhin liu sinh hc Mt trong nhng vn chnh gy rc ri cho cc t bo nhin liu sinh hc enzyme l mt nng lng thp do qu trnh oxy ha khng hon ton ca nhin liu sinh hc ti anode ca cc t bo nhin liu sinh hc.

V d ca qu trnh oxy ha hon ton trong mt t bo nhin liu sinh hc l enzyme cho qu trnh oxy ha ca methanol s dng dehydrogenase ru, aldehyde dehydrogenase, v format dehydrogenase.13 Vi nhin liu sinh hc phc tp nh glucose v sucrose, nhng con ng trao i cht enzyme l phc tp hn nhiu v bao gm c cc enzyme oxidoreductase v cc enzyme khc (kinase, etc) chu trch nhim chuyn ha ha hc. Cc ng t bo chu trch nhim v s c enzyme ca ng l con ng ng phn v chu k ca Kreb. Bt chc nhng di ng con ng ti mt in cc i hi c kh nng c nh chng trong microenvironments thch hp cho hot ng ca enzyme, gim thiu nhng hn ch lin quan n giao thng vn ti vi cc sn phm oxy ha khuch tn gia enzyme v x l thc t l cc hot ng enzyme ca mi enzym l khng tng ng, do , cc enzyme gii hn tc trong h thng. Cc cc nh nghin cu ti i hc Saint Louis ang s dng nhp chut ha hc to ra cc phc enzyme gim gii hn vn ti v pht trin mng c nh enzym m bo vi mi trng thch hp, nhng cc electrochemists ang chuyn sang cc k s ha hc lm th no xc nh cc enzyme hn ch tc trong h thng. Thin nhin pht trin mng li trao i cht phc tp to ra nng lng ha hc t cc ngun nhin liu mi trng xung quanh. Cc enzym tham gia

vo cc mng ny tin ha trin lm mt iu khin phn tn trn thng lng ca vt liu thng qua cc con ng rng khng c nt duy nht trong mng li i din cho mt nt c chai chi phi. Gn y, c nhng tin b th v trong lnh vc bioelectrochemical, nh h thng a enzyme ang c to ra cho cc ng dng nh cc t bo nhin liu sinh hc v cm bin sinh hc. Nhng, cc h thng ny thng c to ra s dng t sinh vt khc nhau m khng tin ha hot ng vi nhau v khng c ti u ha hot ng di trong iu kin in vitro. iu ny c th dn n vic to ra cc con ng chuyn ha ni kim sot ng hc ca h thng l khng tt phn phi, v mt nt duy nht c th thng tr hiu sut tng th. Ti i hc Columbia, cc nh nghin cu c p dng phn tch kim sot chuyn ha hiu r hn v ci thin hiu sut ng ca cc t bo nhin liu sinh hc. Nhng hiu bit ny c th c s dng m bo iu kin hot ng ti u cng nh li xe la chn cc enzyme cn c s dng trong cc mng li trao i cht nhn to. tiu chun ha K thut c tnh Do s phc tp v nhy cm ca enzym i vi vic ch to in cc v sa i, c trng ca cc hot ng enzyme c bit kh khn so snh nn tng cho. Mt gii php c th l s dng cc cu hnh di ng tiu chun ha v k thut c tnh cho php so snh nh vy c tin tt nht. Mt nhm nghin cu ti i hc Hawaii c tham gia vo n lc nh vy thit k v pht trin ph hp vi m-un... cu hnh di ng c th c chia s gia cc phng th nghim tin hnh cng trnh nghin cu so snh to thun li cho vic chia s v chuyn giao kin thc gia cc cng tc vin. Bng cch s dng mt hnh hc thng thng v kch thc, in trng v hnh hc l phn ng c duy tr gia cc th nghim. Cc Kt qu sau c th c so snh vi mt gi nh v hnh hc thng thng v lnh vc loi b s m h

xung quanh nhng vn ny, cc php o ng c th c nh gi. Hnh 3 l mt hnh nh ca cu hnh di ng tiu chun ha c s dng. Kt lun Mc d cc t bo nhin liu sinh hc enzyme vn c tui th , mt nng lng, v cc vn hin ang c gii quyt hiu qu, chng c nhiu im hp dn bao gm: kh nng hot ng ti u nhit t nhit phng v nhit c th, s linh hot ca cc loi nhin liu c th c s dng, bao gm c nhin liu ti to (v d, ethanol, glucose, sucrose, glycerol) v nhin liu truyn thng (v d, hydrogen, methanol, vv); v s dng cht xc tc bch kim khng ti to. Hai lnh vc ng dng chnh m ang c xem xt cho cc t bo nhin liu sinh hc l enzyme trong c th, cung cp nng lng cho cc cm bin v cy ghp my to nhp v ngun cung cp in ex vivo cho cc thit b in cm tay nh (mng cm bin khng dy, thit b in t di ng, vv.) Cc thit b cy ghp rt c th s s dng glucose nh mt nhin liu v gn y tin b ca nhm Heller ang cho thy rng cc t bo nhin liu sinh hc c th c cy ghp v tip tc hot ng trong mt c th sng (mt qu nho) .14 i vi cc ng dng c th c, nhiu nhin liu ang c xem xt , t ru thnh ng. Hnh 4 cho thy mt ethanol 8-cell / oxy nhin liu sinh hc t bo chng nguyn mu hot ng mt chic iPod. S la chn nhin liu ti u s l mt chc nng ca cc ng dng cho cc h thng ny. Tt c trong tt c, nhin liu sinh hc t bo l mt cng ngh giai on u vi nn tng khoa hc v ro cn k thut vt qua, nhng Chng l mt cng ngh y ha hn cho mt s ng dng.

Ethanol / khng kh t bo nhin liu sinh hc ngn xp. Mu th nghim ny, pht trin bi Akermin, Inc vo nm 2006, quyn hn mt chic iPod. Bc nh lch s ca Akermin, Inc