Research Article Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp.

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1 ub Cooo Bo c o oum 216, c , 8 ://xoo/11155/216/ c c Co, xo, Cczo o o moc oouco C om obcu CB1 Xbo J, Jc C, C, Xj o z u, uj cm o cuu cc, uzou 353, C Cooc ou b o Xj ; xj@omcom c 18 c 216; cc 3 Ju 216 cmc o: m Co 216 Xbo J o cc c bu u C Commo buo c, c m uc u, buo, ouco mum, o o o o c C u m cc c () om obcu CB1 co moouco c co comb x cc co ( co), c m o co x o c o u obcu o 1,467 b co c 488 mo c u, c o 81% m o ou u Bcu c m o mo c uc comb c oub co m u 3% o o co o mo bo o comb co o c c u o 35,831 U/m c o o comb Bcu Z-1 mo o c ouco u comb c cc c o 4,526 U/m m o 511 m om co mu o comb zm 6 Co7 C, xb c o o co mu, om 1 Co9 C zm 947% o u c cubo 6 Co1 ou xc moc o ub u o c u co 1 ouco C (C11116) c o zm cc cz comoo o 2 O 2 o 2 OO 2 [1] 2 O 2 b o zo bc oc mc, oo, x, -m u [2, 3] u 2 O 2 ouc o b-ouc mu o um om u, mo o u 2 O 2 c x ouco oc, oo, ouo o, o [2] C coc o mo 2 O 2 u o cc c o co ouo com cmc comoo mo [4] C c b o ou c: moouco m c, c-ox, o-m c, mo c [5] C m,, u, mo obc bc, om obc bc [5] Commc c ouc b m, u, mcob mo m moc c, bu m u co mob c; o xm, x bc mu u o 6 C [6], o moc c mo comb o coo b om moc c o [7 1], bu c-ox o - c moc moouco m c o o o c o zm ouco mo oc comb xo cc mo o c o Cu, c o [11], u omo [12], cococcu c [13], Bcu ub (B ub) [14,15], cc co ( co) [9] u o c o ouc comb c, bu o comb c m moc, uccu comb xo c o moc c c o co m, uc coc,, o uco omo co c Bu m moc zm coboouccommoco[8]

2 2 Bo c o o xm, ou mu mou c cou oub b x co, combc bou c [8]; comb co Bcu omou c-ox o o comoc(1553u/m)[16]o oub xo c om co mo o moc c ou, c moc bc o om -mu como m obcu CB1 ou o b c ouco o, o, co o co moouco c om mo obcu CB1 comb x co oub om comb c o u ccz 2 o 21, Bc, m, um co B21 (3) cuu ou boo, xo co -2 uc om Bj Boc obcu CB1 o uzou, C B mum (1 / o, 5 / xc, 1 / C, 7) u cuu co B21 (3) uo-uco mum Z-552 [17] u o uc xo o comb c mum o cuu obcu CB1 co o o o 9%, xc 5%, C 1%, 2 O 4 1%, 2 O 4 75%, xo co Couco obcu CB1 cub 6 C o /m; om xc cco o mo o Zou [18] m u m C: (5 -C- CCCCC-3 ) C: (5 - CCCCCC-3 ) om c co (C) om x om (,, C) u oo coo: 95 C uo o 5 m, oo b 32 cc o 4 94 C, 4 55 C,1m372 C C ouc u u C uco (Om Bo-, c, U) uc b o Boc Co, (, C) omoo c o mo c uc c ou B (://bcbmo) om b u o z omoo o mo c uc; ou o uc () b o b bo u o b om uc m o b c o couc oc o mu m 42 u o couc oc u m o mo c uc c ou u 6399 u m -2 u cb ooco o ouc om o com co c -1 ( Boc, Bj, C) o co c u 7 omo m (5 -C- CCC-3 )C(5 -C- CCCCC-3 ) cuu B mum co μ/m o mc comb co -2- xc u m (Om Bo-, c, U) om o com co B21 (3) c o co o B21/-2- c u C cb bo 23 uc xo uco o comb C o co cuu 5 m o B mum co μ/m o mc 37 C 22 m o 12 x, 1 m o bo cuu ocu o 5 m o Z-552 mum cuu 3 C22mo16oucxoo zm o com xo, 8 m o uc cuu cu 1, o2mu 3 m B (5 m, 7) u cuu ubjc o uoco 4 u c, 2 μ o uoc m cu 12, o 5 m 4 C, u o cuubumm bo ubjc o 12% - o c xo o comb zm; m co m co u b coo coo m oo: 3 o 3 m, oo b 8 u coom b bu 25 ooco [23] u o 24 comb zm ouco Cu, 1 m o u ocu o co 5 m o Z-552 mum, c b 22 m 3 C 4, m coc z o cc c m uoc bo c mum coco zm ouco cu ob 25 uco o comb C, m o uc comb co cu 1, o 5mocBu(5m 2 O 4, 3 m C, 8) cuo, comb co u 3 m o Bu u m uoc u c uoc m cu, u ubjc o c-mocc c (-) coum, 1 oum o Bu (5 m 2 O 4,3m C, 8) u o o occ bou o coum, 1 oum o Bu B(5m 2 O 4, 3 m C, 2 m mzo, 8) u o mo o o, 5 oum o Bu C (5 m 2 O 4, 3 m C, m mzo, 8) 5 oum o Bu (5 m 2 O 4, 3 m C, 5 m mzo, 8) o u comb c u o c com b - u c c o b z Bu o mo coco o mzo Bu c ou u mzo mo u

3 Bo c o 3 o u c u Bo mo [24] 26combCCcc cc om cco o mo cb b B J z [25] u comb c u m B (2 m, 7) o ub coco, 27 m o 3 m 2 O 2 (u 2 m B, 7) mm o 3 μ o u zm bobc c 24 m mu 5 o 1 m 1 U o c mouozmocomo1μmo o 2 O 2 m c mum om m o om co mu, 3 m 2 O 2 cub mu om 1 Co 9 C(1 C cm), c mu mu bo cub 2 O 2 o zm mob, u zm cub 4, 5, 6, 7, 8, 9 Co3m 1 ou cubo m, zm mm oo c u c mu mo cb bo omum 3 m 2 O 2 oo5mbu:oumc5m ( 4 6), oum o 5 m ( 6 8), c- O 5 m ( 9 11) c u mu bo c c m b ccu c o zm u coco o 2 O 2 om 25 o 25 m 2 m B ( 7) 3 C m ob b oub-coc o cco o Bu [26] 3 u cuo 31 uc C ouc (u 1) uc u o o o 1,467 b, c co o 488 mo c uco uc ubm o B, - B cco umb b uououcocbb o omoo o mo c uc mo uc m c uc o o obcu ; o, o m ubm om o uc om, o b ou co o x mo u c uc ob om o b,cmomoobcu [27], 8% oc (u 2) bomocucmo c co oc o c o Bcu ub cobcu o b u m o mo c uc o c c co mo c u o m b (u 3) o o mob c [28, 29] om obcu omou om c bcu bo o co ou o c m (cox) co b, bu c (b) u 1: C ouc o 1,452- b coo o om obcu CB1 :, m; 1, C ouc bo o ou (moouco m c) c co b [5] o, o ou o u o co x o obcu 32 Couco o comb xo co - 2- o co c, uc com b uc xo co -2- (u 4) om o B21 (3) c, - o com c x o m mocu o zm b ob b 6 7, comb c oub (m u o 3% o o coo) o cuo bo co (u 5) oc mocu o comb zm, ccu u mo uc, 56, cu 6, c coc mocuo65ob-om u [3] ou 6 m u comb o mocu ; o, o o omo m uc cccouc4,526u/m u om coo (6 C, 7) 33 ouco Cu o comb C ouco cu o comb c couc 3 C cuu 3 C o 2, o c o mo bo c mxmum o 35,831 U/m u om coo (u 6) muc m c ouco o o comb Bcu Z-1 (b 1) o ouco u u co o c

4 4 Bo c o u ocu (47623) C uu m (O974923) u ocu (47623) C ocu (64454) uo uo (973) c uum (96822) oxom o (9XZ51) coum coum (O772292) 91 oc b (1) 85 Bcu ub (26915) obcu B1 (JO27321) cobcu (32652) 97 omc zu (33569) omc cocoo (9Z5981) 51 obc ombo (27711) uomo u (597141) 5 u 2: oc o CB1 c mo c o o o o c mo c uc o uc c b ub om CB1 cuc o b; cco umb bc coo cco umb o o b oc couc 42 mo bo-jo o o Boo o co b 1: Couco ocomb co B21 o c (U/m) c comb co B21 (3) 35,831 u comb Bcu Z-1 39,117 [15] Bcu Z-1 28,99 [19] xuobcum oxoo , [2] mcc BC8 2,353 [21] zobum obc ,35 [22] uu 692 [22] 34 u comb C Ccc 341 c o mu o C mu mjo co co o m zm c b o ou u c mu c u o c o o mu, om 1 Co 9 C (u 7()), o mxmum c 6 Co 7 C mu om mu m o c, uc mcc BC8 mu om Co 7 Commuo2 C[31];cobc co -3 om 1 Co6 Commu o 45 C[32];Bcu u BC b4 om 2 C o 4 Commuo3 C[33]c b mu 6 C(u7(b)) cub 7 C o 1, zm m >7% u c; o, cubo mu >8 C, u c <1% cubo o 1ouzmc 8 C, m c o co m 8 C 9 C moc c o om moc mocu ucu [34] c % u ccubo6 C o 1, c m o o CB1 c moc o muc b o o o c, uc o om cobc co ( co)(65 C, 15 m, 2%) [32, 35], bo moc (6 C, 2 m, %) [36], bo umo -1 (65 C, 1 m, %) [37], omo 1 (55 C, 3 m, %) [38] o b zm o b bo o -mu oc u om o zm 6-7, zm>5% o c b 5 9 (u 8) 342 c m o C zm c o comb c u

5 Bo c o JO27321 Cou JO27321 Cou C m JO27321 Cou m JO27321 Cou m JO27321 Cou m JO27321 Cou u 3: u m o mo c uc o CB1 c c om o b uc o om Bcu ub ub 168 ( o umb 2691), oc b uo ( o umb ), omc zu CC 1712 ( o umb 33569), obcu CB1 (B umb JO27321) c mo c u bc co u o o co u o om b oc o o c 686 b -2-6 B 1O m B322 o c 7 u 4: comb xo couco o co - 2- cmc m o comb xo co -2- co c o xo co um B 7 omo om 6 2 O 2 coco ub; Bu o u o ccu m,511m, mx, 1515 mo/m m (u 9), c m o ou () u 5: - o comb c xo u zm :, o m; 1, o bc o o B21 (3) c m co - 2;2,oubbcooB21(3)co -2- uc xo; 3, oub bc o o B21 (3) co -2- uc xo; 4, - o u c u - coum

6 6 Bo c o 4 C c (U/m) m () u 6: C ouco cu o comb co B21 (3) c (%) 9 8 u c (%) mu ( C) 8 5 m () 1 () 4 C 5 C 6 C (b) 7 C 8 C 9 C u 7: c o mu o comb c c () c o zm 1, 2, 3, 4, 5, 6, 7, 8, 9 C; (b) mu b o zm u c o comb zm b cubo 4, 5, 6, 7, 8, 9 C o 5 1 o u (525 m [23] 41 m [2]) o c om co -3, cococcu uu, Bco, cobc o, mcc, Xomo cm m u com o u c [39] o m c zmo 2 O 2 zm b u 2 O 2 coco m mo ccc o zm o m o o co 4 Cocuo, co, x, u, ccz o moc bc obcu CB1 o m comb zm cou m b o c o o mu om 1 Co9 C cc c o u comb c 4,526 U/m o o, comb co B21 c o c ouco o 35,831 U/m zm o co mu, oo mob, ouco m b ub o x, -m, o u Com uo c o coc o

7 Bo c o 7 c (%) u 8: Om o comb c bu u o c o 5 m oum c ( 4 6), 5 m oum o ( 6 8), 5 m c-o ( 9 11) c m 1/ (m/μmo) = 3393x = / (mmo/) 1 5 u 9: Bu o o comb c zm c 5 m o bu ( 7) 3 C com o uo b occ c ubc ( ) uj c [213] o 1464 c [1] - o, uj, oom, ucb ouco o coo ox c c mum b mocu ucu O 31693, Jou o Bocc Bo, o 99, o 3, , 25 [2] - Z, -J C, X- o, -, Z, - B Z, Omzo o c ouco uco cczo o o co- C-2 om moc bcum mcc BC-1, o coboo,o6,o4,71 78,21 [3] o, Bocoo x/co u, Jou o Coum u & om coomc,o 23,o1,53 61, [4] X u,, J o, X Zu, u, uco cczo o c om m bcum cobc 81, Bo c o, o 214, c 49626, 7, 214 [5] B ooc, B u, u, c o mcob c: oo, ouco uco, Bocoo c,o32,o8, ,214 [6] o, mcm o o ox bc, x Cm Coo, o 29, o 11, 12 13, 1997 [7], uo,, uco co o mob m c om moc bcum, c o Bocm Boc, o 362, o 2, , 1999 [8] o, Bco, oo, mu mou c co o ouco o moc - c c o b z c om cc co, om coboo,o7,o7, ,24 [9] b mo, -o -c c om moc bcum oxo ccco, o xo uco, o 57, o 2,255 26,28 [1] uj, O u,, cox om o moc Bcu o o m o x bc u, xmo,o5,o6, ,21 [11] X-, -, J, Z-, J Zo, Zou, - xo uco o comb um c c o, o xo uco, o54,o1,24 29,27 [12], o, U m, comb u omo boc: coxo o c co ox (O) c c (C1), coboo Bocoo, o46, o 1, 46 54, 1996 [13] oc, o, J J oux, - c o ox cococcu c co b Bcu ub c, coboo, o151,o9, , 25 [14] X,, Zo, X uo, Zou, Oxo, uco cczo o comb c c om Bcuub, Bocoo,o3, o 1, , 28 [15]Xu,uo,u,JZou,JC, -co c c ouco o c Bcu ub Z-1, Bocm Bocoo, o 173, o 8, , 214 [16] uo,zou,c,xo,zu, co, oxo cczo o moc c co, c c, o 365, , 212 [17] u, o ouco b uo-uco cuu, o xo uco, o41,o1,27 234,25 [18]JZou,Bu,Jj, coom o o comoo, omcoboo,o62,o2, ,1996 [19],Zu,ZZo,o,u,JC, c o o ouc o c ouco b Bcu ub Z-1, C Jou o om Boo,o14,o4, ,28

8 8 Bo c o [2] o umoo, uo o cuu coo o x xcu c ouco b xuobcum oxoo -2-2, o coboo,o65, , 214 [21] - Z, -J C, X- o, mcc BC8 c o om u co o ox o c c ouco b uo o cbo mbom, cc, o 11, o 1, 37 43,211 [22] m, jm-mb, m, uc,, uj, c ouco b zobum obc 2-1, Jou o Bocc Bo,o16,o6, ,28 [23]uo,o,ub,, mo o o ocm cu occ ocu c bcou om u Coom B Bu , coo, o 9, o 6, , 1988 [24] Bo, mo o uo o mcom u o o uz c o o- b, c Bocm,o72, o1-2, ,1976 [25] B J z, cooomc mo o mu bo o o ox b c, JouoBoocCm,o195,o1,133 14, 1952 [26] Bu, mo o zm oco co, Jou o mc Cmc oc, o 56, o 3, , 1934 [27] Bo b, oo, co co o c om Bcu ub,, o 19, o 1, 31 37, 1991 [28],Z,o,Ju,CXu,JZu, uco o o mob c co, c mcobooc c,o42,o3, ,22 [29] o, oo, O, Co, uco uc, xo cc co o Bcu omou ox (), Jou o Bcoo,o171,o9, ,1989 [3] Zo,X-Z,JC, Oxo,uco, cczo, oc o bo mo, co mmu, o 74, o 1, 61 65, 26 [31] - Z, -J C, X- o, Z, -B Z, ouco, cczo, co uc o moouco c om mcc BC8, Jou o Bc coboo, o 51, o 2, , 211 [32] moo, omu, um, umoo, Cczo o c om coo cobc co -3 xb c c, o Jou o ocu cc,o13,o2, ,212 [33] Z,X,X,Zu,C,Xo, Cczo o c-b c B o om Bcu u BC b4, o coboo, o 66, o 1, , 216 [34], ou, om, uj, Um, uco cczo o mob c om cuu bo o mocu ucu, Jou o mo Bo, o85,o2, , 1998 [35], u, u, B Z, uco cczo o coc c om cc Bcu, C Jou o coboo, o 54, o 1, , 28 [36] oz,o,jou,, Co u o bo moc c: cco como o moc cou om ou mb, xmo,o1,o5,427 44, 26 [37] umoo, c,, uco cczo o c om cu coc bcum bo umo -1 xb c c, Jou o Bcoo, o182,o7, , 2 [38] uco, oo,, oz, uco, cczo, uc o c om - o-o bcum, omo 1, Bocc, Bocoo Bocm,o66,o5, , 22 [39] J C o, o o mo c, c o Bocm Boc,o41,o 2, , 22

9 o Jou o Bo c o ub Cooo ://com oum 214 c m C o ub Cooo ://com oum 214 ub Cooo ://com oum 214 oo ub Cooo ://com o Jou o omc oum 214 ub Cooo ://com oum 214 Jou o ucc c Zooo o Jou o ub Cooo ://com ub Cooo ://com oum 214 oum 214 ubm ou muc ://com cc o Jou Jou o uco ub Cooo ://com c c o ub Cooo ://com oum 214 om c o ub Cooo ://com oum 214 zm c c ub Cooo ://com ub Cooo ://com oum 214 oum 214 ub Cooo ://com Bocm c o o Jou o coboo ub Cooo ://com oum 214 o Jou o ouo Boo oum 214 ub Cooo ://com oum 214 ub Cooo ://com oum 214 ocu Boo o ub Cooo ://com oum 214 c Boomc ub Cooo ://com oum 214 Jou o Boo oum 214 ub Cooo ://com oum 214

WAS PICKED Bt PRESHENT WILSON NUF. '. a ji.,.v... ;... and ' ' V. -, T.. TL PfVlfH A M. u ; Moersberger, Lieutenant. .;: r 1...

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