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Biofuel Enzymeキット - Bio-Rad

1. Diane Sweeney Biotechnology Explorer 1
2. V A pH
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5. 1 2 2 5H H
6. pH
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8. 0 1 1 1 2 1 1 MERE 200 mL l 1 200 mL l 100 mL 100 mL 3 mM 100 mL l 1 5 mM 150 mL l 20 mL 5 4 450 mL DBTP 2 1 000 mL l
9. p SEE M 1 5 mM Ix epn keres 1 5 mL DPTP SS E C LME L DPTP ds 1 2 1 9g wW N 76 YE e e e
10. p pH 5 0 pH p pH p
11. AE SRM ccr EID SA eR Sd um pecu OUI UEUPE UN ses d 7 cce 8 25 1 1 5 mM 15 mL joo E1CES5 2 TK f
12. 37 EE Rei 6 2 B
13. DPTP 100 250 500 750 1 000 uL DPTP 1 mL DPTP 6 10
14. 3 3 e e e 8 2 H O Ho HO 1 4B 2 3
15. 1 5 mM 120 mL 40 mL 120 mL BRAERE 37 y 1 64 mL be etry S1 S5 8 mL 9 32 SS 40 MEMEA VCI RORA T i 1 1 1 15mL 8 1 5 mM 1 5 mM 15 mL 2 15 mL 8 5 mL 3 15 mL 8 1 15 mL 4 15 mL 8 1 8 mL
16. F PALCOM CIFA ENDT a Ae
17. jc c 95 C 3 5 A 3 mM 1 5 mM Lineweaver Burk 8 6 24 10
18. 20 40 C Groudieva 2004 1 5 mM DPTP 1 ST SS T5mL Seco KREAN E S RCEOND AO e eese DPTP 1 10 C 122 C 372C
19. Arnold and Georgiou 2003 Zymomonas mobilis Escherichia coli
20. 8 C100 859 ESS Amo qud Rod 88 C Bio Rad SmartSpec Plus SmartSpec Plus SmartSpec Plus SmartSpec Plus Av57 1 SmartSpec Piuns
21. nmor 0 1 JY wUJOAZAgUH 12 5nmol 2 d 7e 4o 50 2W 2E XE fir AU 29 96 9 Hy MEERCCIS RO REIX RE C OEV ORRI ERRED EEVI ETT WORF T XAEBURIE TET FAXO C KIREST TE ERERKEN RN ET 2 l DIREI 07242 E OUKEOEE T2 09 CHo TROET ATAEBEB EB AIRO FIIOOARODR C I OB EEI37C CA bJ f qdX2cd Om C EIKURIE NISI Cd JE REDLEDU UJEeXtcs sEmRD nuEeZS t itHuiBuEecFH mOn LAE BABEIEJJUfI amp 2 DICBEHCAIFIHICEA2bLE VA
22. Biofuel Enzyme p wu pU Josie psc Bot 7 LSND c LE Bd 1 p p p 1 p p B 4 C 1
23. EL CO 1 EE P 410 nm pua SR E S PE E o P nmol 18 P nmol 410 nm Em E N H D LI L P 19 2 0 C 0 nmol 95 C 3 iB HLic
24. 1 6 410 nm 5 17 2 2 1 p 17 3 17 p 8 P 410 nm 79 1 P nmol 2 nmol 6
25. SI 9 68 5 Varro diei 2 ON A E 6 iis i ANO MM au ced QM AL d
26. 1 pH 2 Efe Didnt TENCE D L TRER ORREZ ESI LICI
27. 93 G L BEC CHAR Ti HER5ISZUDEATCRS H BHEROIDSSEbbUET A22 AO RESLO Jue ORRE FERAT SOSuk uemuELEYd BEDIEF D KREA ATE HAm ZITI I LRAELT FEDERE D SOET MIELE ENDT SZ BEOR ED MROR ORALI SZETT HANDOG EE ofm0HBRLEDSO INETIBRUDImY BNEHDOEUPBAEMDPOCS CDOT IZRAELA BE HADT OMERO EERIE E TIFE e UEFI EIES EET MUI TRILLE Elt HARDT EIR T BGEIIEREEKBREZAUB 4LUL SGREKRRATYOcrIquex Am edrFedE CAIC KEH
28. 3 19 1 3 8 Montenegro 2006 Martin 2008 13 86 p Am CO M Ly wf ay Eos 25
29. AIC LU SEE e 1 2 3 pH 4 e 5 e 6 ede
30. KERALO SILT KA 2 R RT A 3 7 3 I pH TIBI EREE E HE BIO SEIRTIRIES COD IE BE RR T OB EEIBTRIS CE ILE Cd Ef HIER OO ET ODER OR ECTE DV ET pH COOBESOSRuiBpHIApHS5Xr pH 5 0 CRI AE RC UR E 2I Cdbd ENIF ZaD TOEI PIECE pH pH
31. 3 2 2 1 4 2 p p B 4 2 p
32. 1 f e A aM e e e e Po E N e EnF 3 LI pH e e DNA gt RNA gt gt e enid TYFORS RID Em 1 P T 2
33. DPTP V VW VU OCE OCS 22 CE 22CS 3 CE 37CS 1 37 C 0 C 0 C 22 C 22 C 37 C S E S E bes 2 29 3 pH 1 pH 5 0 pH 6 3 pH 8 6 2 DPTP 200 uL DPTP pH 5 0 pH 6 3 pH 8 6 3 DPTP 3 0 mM 250 uL IpH 5 0 pH 6 3 A pH 8 6 DPTP s j wv 3 0 mM pH 5 0 pH 6 3 pH 8 6 4
34. 54 l S81 S5 5 0 C 22 C 37 C 5 6 5 p nmol 1 1x10 mol 0 000000001 mol 6 p 3 p Bonoa nmol Lo 0C 0 o 22C GER NEMEEM NEM NENNEN NEMMNMEMMZNMN 2 1 S1 410 nm 3
35. ET CEA Z EOHR s BHHEBZeeAIC B Y 1 ISEUH EP n RREZ IL ODER 2 AIXS2EFubr BUlck04d xsseb cO4u d DIBSMEIWEOTSfE FR CHOICE ET ZJDUESETHHflcd0 comBmBBONcez BBLamBOALueSREI ENTL dSL WES COUR QEEY 94 9 FERIOBJHELTUCEKETLZS HUEZZTZ GHLTOQUIEAD ERL TOSHWHUIO EG dba ldo ERHBPIOIER IERI LEY ER CORIR YEDE QOO ElkioOu VO LORA CHI TROS ERTZE INIZI nmoles 2i fir COUR RE 23 FONET 95 1 L bx lem ZcbSecCIwESAs PHEIG SIC HQ OAS LH EZGA B
36. PR GEH 2 6 2 B1 4 2 p eun pre Ah Ge Dsm 2 0 1
37. 3 4 6 GM 2 2 4 2
38. VQ K A 15 1 2 13 410 nm v 1 5 mM 2 mL 1 75 mL o GRE 1 5 mL g KIER 1 3 25 mL g S1 S5 1 o 53mE 2 DUET ect 2 g DPTP 4 o Ak 2 Sees edes ur l J DPTP 1 o KN o 6 45 l 15m
39. erg e ROT S 0 f amp T 2e 38 o C v ARMER E ZIHER dod 0E nU OIXORSEDA ROO KERMAT 5B RRO 3 2 1 2 6
40. p HH OH Do BEDANTA p OH H O A p HHZ2rz Z WAab6222F OH 9 O A yY lt B Sus X CEN um u P sid o 07 E ro O p 4 A p
41. 9 DPTP 250 pL 3 mL 15 mL 10 15 mL 500 uL V 2 Co 3 4 4 6 5 1 1 2 2 4 3 6 4 8 5 78 11 DPTP 500 uL 6 DPTP 1 86 12 DPTP
42. 15 8 nmoD nmol 0 1 5 2
43. 1 98 mL 10 20 1 5 mM 1 5 mM 3 mM 75 mL 1 75 mL 1 1 Bt 11 WE 2 100 mL 100 mL 7 a 50mL 5 S81 8S5 b 10 mL c SS5 4 mL 6 mL d S4 S 10mL e S3 S4 10mL f S2 S3 10m
44. IUUD TESI eux mal BERRE CIERRE Xd L LEEXZIEMA BRAMZUE4 WA REFRA CERE dE mE IROBESGC UHY cs EOL I RRT QUE CIXZ IRIURE efj5o4v5Xed cossd ee rocs amp cy WALER ENEJ Z e IRCO TEED h ARE TOOT ERUETI 99 l 5 m EmOMEGC QEILIOEC GERI LALESY BXLPBACEDDUPIES Oc BARE MX DA5 ZEQZUXd EMERE C E IE CEPA z Dh VFI TORA QC Z74 FAT BRAMEHE C4 SAER RICOR TOWE ODEA ROERE IE DOCS REI IO 5 SFIN EDET E EERIE LL de OUB VZAMEB 5Xoitzedc LvX XSTX4 Er BIdBSoLXb5 AU E AEF RTJIBI X EE livw JXUGEHIqd4EnHIQCIBERERSSL Xd BEubr PCctutktugd xeUllhad bI SCOUET KIRO Es RENE ChbsZuu Xxb Bzrz 100 6 b
45. 7 DPTP 750 pL 15 mL E el 8 DPTP 750 uL 15 mL V 71 9 DPTP 7 15 mL 200 uL Sess RICE S od d U a H1 1 H2 2 H3 8 V Ji LE L1 1 L2 2 L3 8 H DPTP L DPTP
46. pH 1 Tiago etal 2004 pH pH 1 BE CU956004 PCR
47. 1 S1 410 nm 6 16 2 2 1 16 3 16 p 8 P nmol 410 nm LUE I I OO MEN e O EE CE II ci o B EI e a a MI 73 1 1 Eat E E nie s N H D LI L P 2 0 0 nmol 1
48. 2 Sk Lx 1 p E nmol 1 1 x 10 mol 0 000000001 mol 45 2 2 1 S1 CS 3 1 S81 410 jm 410 nm 3 p 7 Jvi amp X 410 nm 3 Sn P FA7Ir V T 2 EI E5 410 nm 4
49. 3 mL 2 13 OE 410 nm 3 5 1 v 1 5 mM 750 uL g 3 mL o BRAS 7y 1 1 25 mL g KISER 1 3 25 mL g S1 S5 1 o DPTP 4 o 150b c US TS 1 g 6 g 1 g DPTP 1 o APS T PRA S pote 1 16 5
50. 33 EPA E R OAIE AO E 35 ff 37 42 DIREI NEOA OS 52 pH kk 58 kk 63 kk 69 ETARE E OIIE AOL E 76 DE r1 EE our mE EI D RENE 82 FAB AA A A F n i 86 C SmartSpec Plus 89 E F as 96 G lt 98 H kk 106 Biofuel Enzyme 8 p p
51. Sot HABRA CLA D Ala a eaa BA pH UH BL RTR oP EF 2 RORE SIVCOBPESHIAIRSE D h DIRIR ET OCRA ALE To ROER CIEBES TI eL ARAS Sv L fBoUPnELFEE2 OBEDOM J DTT GUHJH 3 D7r ONTLUV Ur OUDLRADNEE 5 PRETE b EREL FEF duiiueBtkize6t2ffwcwe6ugettbeeecd TuBfkssE e amp erbLt6b5ccLr 5 pH
52. OCA Syngas synthetic gas
53. 1 IS1 S IHIeH3 lL1 L3 11 12 11 p nmol 1 1 x 10 mol 0 000000001 mol 12 p p ER 2 1 S1 410 nm 6 13 2 2 1 p
54. RA 20 40 C pH RDN
55. E IIIRZ TLE ORR CAINE D VMESIX Eik CIH LE bo X VERD EL EENT DOCERE Roe NVI A C OOfUHl EGRDYmENE TEM LEBEL VE Wi CORRES EOL OJE coh i2kBe CIH LERRA L V REH CTCORERED OLOJA D Ux OI 9 pH Bli o KpH CR 5feUEH I E RICHIE FORRAR RAR EERTE ROET HEERO HHP W DOL ATERAT S SJBEZCTOdwcec8DAE 101 H Arnold FH and Georgiou G eds 2003 Directed Evolution Library Creation Methods and Protocols Totowa Humana Press GenomeWeb Daily News 2008 Genome of Termite Gut Bacteria Sequenced http www genomeweb com genome termite gut bacteria sequenced Accessed August 24 2009 Groudieva T et al 2004 Diversity and cold active hydrolytic enzymes of culturable bacte ria associated with Arctic sea ice S
56. 1 2 R 3 57 3 pH 1 1 pH pH pH
57. 7 DPTP 750 pL 15 mL S W 8 DPTP 750 pL 15 mL J V 9 DPTP 15 mL 500 uL 029 ll 10 J H1 H2 H3 DPTP 1 2 8 V L1 L2 L3 1 2 8 BE 34
58. TCT AA OFERE AULA 2N8Vw 7M EUER ES D OC BERIETE KLE Z boR BANDT R E T LERO C ZSIBEC MED ZNRTIMEARBBIABEILEy DOZ FARERI UR EE OR NE A OCE Fe JGEOBGEI T MONZE ECS Blvoz HNmBiCAHLCOLEwEd CUIT S 2 BROJI KE GQAMBEL ELSZESYO XKduke ofe cBHbnRE m CIKKRE 4v ZWPFYV MAAG SORS O TREEREN ET hikockwizWozk KO BASDMEqVWOICIIAREXEd HAMIRO ET SCRIE RID BHARA T T e OS RERA NM SGOeC C GNE CH DLODBEGEI3TCCOhANeEAOIXSUEZLE L2L amp 8 mKeome ru SOIC WEET LIVES 97 7 IOBEEIEWIAUEC Sf G Opi RACER L TOZ ARSED V ET 1
59. ee Amm X 7 3 g U g U g 0 C 0 C 22 C E 22 C 37 C 37 lt C 6 DPTP bin 250uLZ 10 C DPTP acr e ME MERE LDPTP fii o C37 Co dE E 3TPCeoB3Gm ibuw 37C0OUz 2 5AUCERUXY AhkNv RA a 37C 7 2 DPTP 500 uL 1 5 s I 2 y g UM 0 C S E 0 C 22 C S E 22 C 37 C S E 37 C 8 DPTP
60. E Nut d aida ICT R pH iim db 2 IRIS OO TER SESB LL
61. 4 C 4 C 1 Re 4 C 3 Biofuel Enzyme 8 1 4 4 C v 4 C 400 pL 1 o p 90 mg 1 o p 1 mM 4 mL 1 o
62. 410 nm 6 3 0 mM 5 1 1 5 mM v 3 0 mM 1 mL o 1 mL o KIER 1 1 75 mL g S1 S5 1 g 1 mL pH 5 0 250 uL g pH 6 3 250 uL pH 8 6 250 uL g DPTP 4 o TARP 3 1 g DPTP 1 o A ES EN IM e l o 15 4 45 L Elorri mL IE a ARI
63. 7 2 1 nmol 7 3 X7 3 p 8 410 nm EREE Ho Eu TN nmol Lo 22C GRO LOO U 55 Show S3 0 3 22 nmoll wit AN xs l W nmol nmol 1 2 nmol 0 3 25 nmol a p a 410 nm 25 nmol 0 nmol 2 0 12 5 nmo1 4 22 nmol 0 nmol 2 0 11 nmol 77 p 2 l 1 p B GS
64. p p pp p 1 pH 38 E p p Bl7 p p b 1 p 1 p
65. DPTP 5 092C 0 C 22 C 22 C 379 C 37 C 270 0 6 5 28 6 DPTP 02C 250 pL 0 C 22 C 229C 37 C 37 C 37 C 2 DPTP 500 LL
66. 2 l 1 BRAS Z7 1 10 50 mL 450 mL db xg 400 uL MERE 600 L 1 mL 1 63 mL 20 mL 1 60 mL 3 mM 3 mM 1 1 mL 1 mL MERE 98 mL 1 1 mL 1 mL
67. 13 84 E o E e q 0 01 1 S nmol 13 Lineweaver Burk Ka Vnu Y V K Va CK 85 B 4
68. DPTP S00 uL DPTP LEF 220515 mL 1 1 DPTP 1 5 mM 2 mL 15 mL DPTP T L5 mM 1 mL DPTP 1 DPTP Enzyme E 1 Contro IC E DPTP edu Tp DPTP gt M El 7 s zx s Y UJ 1 5mM 26 8 11
69. jac V 27 m E V V E U EL ES 6 8 E1 E2 1 2 4 8 2 1 02C 229C 37 C 4 2 DPTP 300 uL s DPTP V d 0C 22 C 37C 3 1 5 mL 3 09 C 22 C 37 C 7 DPTP T R250 uL DPTP 4 1 5 mL 3 02C HJ PCERE IS3PCEH 2 DPTP 1 5 mM 500 uL
70. 13 3 66 13 p 8 410 nm dL MEE nmol 1 1 1 gt P nmol 67 2 0 0 nmol 1 1 1 nmol 1 1 nmol 4 1 2 3
71. pH 3 pH pH 7 2 9 0 3 0 mM 1 1 g 1 1 SI SS 1 g 1 5 mL pH 5 0 pH 6 3 pH 8 6 1 DPTP 4 g 00 p 3 1 o DPTP 1 g 1 g v pH 3 0 mM l pH 5 0 IpH 6 3 pH 8 6
72. du DPTP V o 6 36 vB re elite DNA NUM iMd dM AM CR ME
73. 2 2 2 3 BRT Anni 3 92 3 2 1 4 3 2 1 4
74. 2 A kh S 1 5 mM Iy Rd e 1 S1 S5 1 151nL I F DPTP B gus ca a DPTP AIMO SIE c aT sues UU 0 A t kh S 1 15 mL 1 15 mL 1 2 DPTP 1 5 mM 1 mL
75. Smith 2009 L
76. 1 1 nmol 1 1 nmol 74 5 1 2 3 75 6 1 4B
77. 1 1 4 mL 0 1 0 85 2 8 6 4 mL 2 mL 1 1 mL 410 nm 410 nm 1 0 pg S13 410 nm p 400 4
78. 3 10 2 1 nmol 10 3 10 3 pH p 8 P 410 nm nmol pH 5 0 NENNEN pHes pae 1 3 pH p 1 2 0 p 0 nmol pH 6 3 2 410 nm 0 14 82 p 12 5 nmol 1 0 14 9 n
79. 7 200 uL DPTP M E1 E2 E3 E4 E5 5 15 mL 2 1 1 6 DPTP 1 5 mM 2 mL 15 mL DPTP 1 5 mM 1 mL DPTP TQ 7 Mg es A 1 5 mM 43 7 1 DPTP Enzyme E 1 Contro C E DPTP
80. 8 10 12 C DPTP 300 LE TAA FEAN CS ML NP 500 uL IE DPTP 1 mL 15 mL E DPTP 500 uL C DPTP lay psn 500 L DPTP
81. 8 S1 SI 1 mL 8 S2 S82 1 mL 8 8S3 8S3 1 mL 8 8S4 S84 1 mL 8 Ss S5 1 mL QU eS NSP 1 3 13 1 15 1 2 13 WE 410 nm
82. p 4 p bx ES 0 00 2 46 3 p p p 410 nm p p 8 9 5 0 BEB3 0 73 y
83. 3 ta EORR IY P TN 2 1 Lo M 8 NII 50 410 nm P nmol Tig 2 S H D mi L I o B 1H 6 23 1 4 mL D gt 0 2 1 7 2
84. iE LC 57585 04 00p ha 7 x 7 viIX0 nmol 3g QUOTE SOT 3 E def oC Th E AN B EH BI 0 C 22 C 37 C l t e C v nouw xig za TEON d 56 2 1 2 C 3 4 5 6 CC 7
85. pH 39 1 2 3 4 5 1 40 6 7 8 9 4 1 1 p p
86. g 2 3 OOV Node Eyes RMY 1 l o 4 6 eccl 4d ec bum 100 1 000 pL 100 1 000 pL SmartSpec Plus 170 2525 UVP DOT SA DyNA Chill 7 ev den eese tt E palo oo cO p v o o 6 30 3 1 2 e 2
87. p uU Gua Oc B p pH5 OH pH
88. RRRRRRRRRRRRRRRRRRRRRRRRREEEEEEEEEMEMEEMMMM 4 SEE LEE CUTS C BU Lecce totins Eu Rai 11 11 cu cc Mg Loy 13 1 i 14 2 kk 14 ETKEN aE o a OAO S 15 4 16 5 RR 17 6 kk 17 li 18 SG 24 1 2 3 4 5 6 dte EB Ed 1 2 3 4 5 6 26 28 pH kk 30 kk 31
89. C DPTP 8 11 8 C DPTP 500 uL 15 mL 500 pL TT EN TT LE Ga 7 7 T T 9 E DPTP 1 mL 15 mL TTTTT fil i TITITTTTTTTI v V 7 Bx 10 IEB DPTP ERMID T m 7 25 5183500 pL
90. DPTP 8 15 mL 1 DPTP 3 mL ar s S E 1 5 mM 9C10 9 DPTP 250 uL 15 mL 10 00 uL Fg d 11 DPTP 500 pL 6 DPTP 1 VJ V 1 2 0 1 1 25 12
91. S 3 A l2 A B C D
92. T DPTP 15 mLa Zi v a 7 6181500 uL T J H1 1 H2 2 H3 oo Y 4 1 V LE O L1 1 L2 2 L3 8 H DPTP L DPTP 1 Hl Ll 2 H2 L2 8 H3 L3 65 8 DPTP 2 6
93. j l Ei 1 E2 2 E3 4 E467 E585 44 11 C DPTP 3500 uL 8 0 6 6 6 6 NENNEN 4 1 12 DPTP BE l S1 CSS 1 7
94. kh S 1 5 mM 1 NNW UE DPTP 5 mb Et 55M eem SIS 1 PES qp CO a DPTP RS ZI NDN e e emee nm Qu a Eaa a a r LI 1 kh S 42 1 RIE 1 5 mM 15 mL 2 5 EI1 E5 3 2 4 DPTP
95. 61 A 1 2 1 Vma KS 2 ER SR E Rec MEL MEILEN oo e dod DIC
96. F F ES ES E 4 5 F S 65X k k k S TA LA ES V iria E 6 Ve wv B O max k Ky kj S 8 UOS TEATATI V M S Kn S K K 9 K k k k Vj K 12 Vi Y X E Lineweaver Burk
97. pH 9 5 15 Biofuel Enzyme 1 i ON NA OS OS e E iiie a along
98. 0 73 s ER a 44 nmol LE EN 3R EX G E c e M P nmol 8 p p S1 S5 47 3 9 47 B1 ES 4 410 nm P nmol 9 p S1 lt S5 p 48
99. 3 H1 H3 3 L1 L3 DPTP 300 uL DPTP DPTP1 H DPTP1 L N ES Wi 1 VJ J U H1 H2 H3 L1 L2 L3 3 S 7 96 6 H DPTP 1 5 mM 250 uL 15 mL L DPTP 1 5 mM 250 uL
100. 1 2 anl We B 10 p nmol E my N H D LI uL P 10 n 0 8 y x 100 nmol 12 5 nmol 8 1 12 5 nmol 49 2 BUSQUE 2 4
101. 8 mL 2 13 410 nm 3 v Ae c 1g 1 5 mM 3 25 mL buie IE TR 3 25 mL 5 mL Im EN Sm A HH v ue DPTP xc d DPTP LA pua pe dq ete Du ILI e Qd uuc ac uL uL LLL LLL e e e me 17 22 C SmartSpec Plus
102. 58 2 DPTP 200 uL DPTP GE T ATTA o i pH 5 0 pH 6 3 pH 8 6 3 DPTP 3 0 mM 250 uL IpH 5 0 pH 6 3 pH8 6 Me 0 0O ODDPIPISZSC 9i5URU SE 95 3 0 mM pH 5 0 pH 6 3 pH 8 6 4 DPTP 250 L Pou ML m En M ec W 3 2 pH 5 0 pH 6 3 59 5 2 pH DPTP 300 uL pH 5 0 pH 5 0 pH 6 3 pH 6 3 pH 8 6 pH 8 6 6
103. vins ditio pine HEB PRIK vieni 5 12 e RS E EN r3 E E R 1000 1500 2000 2500 3000 nmol 12 V 82 V m Ra M V Vas Vo SD tr NG Km S
104. 3 L1 L3 3 DPTP 200 uL DPTP 799 00i V H1 H2 H3 Li I5 L3 4 DPTP1 H DPTP1 L S gt 7 5 IH DPTP 1 5 mM 250 uL 15 mL CTTITTTTTTTTT FEFCTTTTTTTTI v 1 5 mM 64 6 L DPTP 1 5 mM 250 uL 15 mL V V VJ MJ 1 5 mM
105. 52 1 1 l 4 1 o 6 3 1 1 1 l 1 1 1 2 DPTP 500 uL DPTP TTTTTTTTTTTTT FEETTETTETTTI 0 C 22 C 37 C 3 Lsm 7 uT an7 MPS a 22 C 37 C DPTP 220 uL DPTP mLIF T oc EE 3k 22 C E 37CH 4 1 5 mL 3 0 C 22 C 37 C DPTP 1 5 mM 500 LL DPTP 1 5 mM 0 C 22 C amp 8 37 C 23 5 TOCHER 0 C e a 222C 22 C 372C BPCHE
106. ct SR G H o 2 PE
107. Vo Vmax K LL I Ka 1 Vg VON max Vias i S Km S 29 s V 172V K K S K K E
108. 1 2 9 3 pH 60 4 pH
109. 3 2 1 4 3 2 1 4 3 2 1 Kies
110. Bacteroides succinogenes http sci waikato ac nz farm content microbiology htmL B 7 05 Of DI LI UXegcwEkS AREA TCO b Aspergillus nigel tI F E ZEEL TORM 1 IER icc ied Miro ls SEE CUAL Meses TUIJERMC IU UDSDOTEDIUCUNE
111. 1 1 1 ERIS Peggy
112. 12 Hl L1 2 H2 L2 8 H3 L3 10 DPTP 2 6 l S1 CSS 3 IHIeH3 lL1 L3 14 15 72 14 p nmol 1 1 x 10 mol 0 000000001 mol 4x15 p p c Holds iig D 2
113. P 11 E S Dig N H D LI a pi 11 E nmol nmol 50 1 L A
114. Enter number 1 3 of wavelengths to read 1 1 Emter 2 Enter wavelength nm 410 Enter 3 2 Do you want to substract background reading NO NO AX NUL Enter 4 S1 0 nmolp Read to read absorbance Exit Assay gt Options Read Blank A410 0 00 gt continue 89 5 S81 S2 ICE A410 0 230 Samp 7l Read Sample A410 A410 25 nmolp
115. S ES kj k px P E k ES k k E SoESOE P k 1 ES ES E VL Gu US 83 x 3 k E S k ES ES ES kj k y ES k E S Kt
116. X LPHO LoT BRE KEPER o TORAS SNB AREER EN ET n S ipH2 5 EFICAZ GRAXTpH TIIRA ORME B UpH CHUEO SH A AEOOH 4 ZOET o fbtidm3oweR NXI GeVEORZ SO B FEEROSSBECIZCOOH C H AUEDSRCHCOOICZU EZR ICNBG MOTOS T 3 7B H PEOSSBSRCIINH 7 pH NIL TEHO ES VEEGE c P AE d BIRRA Lr NU BNOSOISLAE S EHE DE 2 ZeB FEOESBZ2NU LOwAHdcod 98 4 l ZNATE BROTA BRER DIE BEXEIBZXSMElIIRG O XC WHERE LELAYO CImE OV ZE BLDIREDNTEIRIIIRIEBC V IRAE LX MERR IRE DREA ERRIBERRI AX DET RENCI CEREC ET
117. 1 mL cS lg DPTP 3 DPTP 1 23 mL 15 mL Pis NM 1 5 mM 250 uL 15 mL DPTP p uS 4 H1 lt H3 IL3 CEEGURE 5 DPTP 300 uL DPTP 6 DPTP1 H 1 L 1 5 mM 7 g V V 1 5 mM Lind H1 H2 H3 L1 L2 d 33 7 9
118. ea Halo NE pala S UU 3 1 2 mL mL 1 5 mL 1 5 mL 2 230 pL 1C6 DPTP 200 uL DPTP 8 15 mL 1 DPTP 3 mL E E E U 1 5 mM 10C11
119. v 1 5 mM 3 25 mL g 1 25 mL g KIER 1 3 75 mL g BRAAI Z7 BRE 0 75 mL 3 DPTP 4 o oanloet PST Enn 2 g S1 S5 1 g 7 g 1 g DPTP 1 o APAT AAT ENA ue 1 g 2 30 1 8 2 37 C 8 3 13 E 410 nm 4 1 5 mM 1 75 mL 1 mL 1 1 75 mL DPTP 4 S1 S5 1 5 mL men Cem eS d UNTUEE 37260 KK DPTP 2
120. 6 1 2 1 3 1 2 mL 2 mL gx2 mL 4 5 1 5 mL 1 5 mL 2 iB 2350 L 6 1 lt C6 y N t 35 7 DPTP 500 uL
121. 2x 100 mL 1 o ERA y Z y 10x 50 mL 1 g 50mL 1 o DPTP 40 o p3unbp 7 BT u 90 g Sb 2 50 g SnL TRAY 1 1H 1 v 1 00 mL 1 g 1 200 mL l 100 mL l 100 mL 1 o 3 mM 100 mL 1 o 1 5 mM 150 mL 1 o 50 mL o 4 450 mL 1 000 mL o 1 o AR 6 1 o 4 v so rese uf 1 g 2
122. C4 87 2 C 300
123. 1 B 1 4 http www expasy org enzyme 3 2 1 4 I ERETTE PR ZARI 39 1 4 B http www expasy org enzyme 3 2 1 91
124. 13 C4
125. 15 mL DPTP 15 mL 500 pL du DPTP V 1 5 mM 1 5 mM 32 H1 12 L1 1 J a H2 H3 2 845 L2 L3 2 8 5 l E MM 15 mL 1 A Ici UA 2 DPTP 1 5 mM 1 5 mL
126. 15 mL DPTP J 1 5 mM To 69 3 DPTP 1 25 mL 15 mL DPTP 1 5 mM 250 uL 13 mL DPTP 1 5 mM 4 H1 H3 L1 L3 5 DPTP 200 uL DPTP Ombe T MM Mi H1 H2 H3 LT e B 6 DPTP1 H 1 L 70 7 9
127. 20 ui Qd uucuc uL uL LL LL LL UJ QN 14 3 pH pH 45 1 1 5 mL 8 3 0 mM 3 0 mM 1 mL 2 1 3 mL 8 pH 3 0 GRE 250 pL 3 15 6 lpHSOgRE57y 220 pL 4 pH6 3 1 1 25 mL 3 75 mL 9 IDH6 3 pH6 3 230 pL 5 13
128. 5 A 2 B 1 4 B bo MA C 2 D B1 4 2 2550582 05002 RIT 2 2
129. 6 S2 5 S33 S5 410 nm nmolmL 90 E B 2 B 1 4 B 1 4 cv1 4 2 B 1 4 B B1 4 2 B1 4 D
130. Biotechnology Explorer Biofuel EnzymeX v 166 5035JEDU 4 C 214509 0 1839
131. e 1 e 2 e 1 1574577 e 15 50 e 1 45 e 2 5 pH e 6 30 90 e pH 90 e 90
132. Biofuels West Sussex John Wiley and Sons Ltd Tilman D et al 2009 Beneficial Biofuels The Food Energy and Environment Trilemma Science 325 270 271 Tillotson JK 2007 Introduction to enzyme kinetics assay of B galactosidase Sci STKE 394 tr4 U S Department of Energy 2008 Bioenergy Research Centers An Overview of the Science Bulletin DOE SC 0104 Yu C 2004 Enzyme Kinetics http www wellesley edu Biology Concepts Html initialvelocity html Accessed October 14 2009 DyNAChill Labnet International LoggerPro LabQuest SpectroVis Vermier Imternational American National Can Company 103 S haa PB 116 0014 2 2 24 Tel 03 6361 7000 Fax 03 5463 8480 532 0025 1 14 11 Tel 06 6308 6568 Fax 06 6308 3064 812 0013 2 5 28 Tel 092 475 4856 Fax 092 475 4858 Tel 03 6404 0331 Fax 03 6404 0334 Mail life ps jp bio rad com http explorer bio rad co p M10504 1303A 104
133. 9 1 60 80 2
134. 10 nm p 4 C 1 1 3 pH 9 5 37 C pH pH 8 6
135. DPTP pH l S1 lt SS pH 5 0 pH 6 3 pH 8 6 pH 8 9 8 p C S nmol 1 1 x 10 mol 0 000000001 mol 9 p 3 pH p P ee nmol pH 5 0 a pH 6 3 a pH 8 6 a 2 60 pH 1 S1 410 nm
136. DPTP 230 uL IDA pH 5 0 pH 6 3 pH 8 6 N La V y 3 F 7 5 2 pH f gy DPTP 00 uL T 1 T 6 B ibd pH 5 0 pH 6 3 i pH amp 6 l DPTP 2421 30 4 l 15 mL 1 DPTP 1mL DPTP 1 mL RR DPTP
137. E n PIA ER AICO ET MR LIESULSoCCOoSRBEMEeUGES COAT XL Edps BIST mg4evpe EZB ERSUISqQqw EC Wb wp H7 AAEL ET RIC REDRE ENCORE STM IC bR TRECERII ON ECA BEXMEXqc Ski cd 827 f 22100 nmol C3 70432 5410 nm COR O REZ3I 60z27208 0 ET 13 8 CAVETVODEFIBIBIS CAEBCIRTAJB X Xd PUHCEOIBmDUROn5m itmeiEc EWEl iE ER OCAuSIVOLTAGEY COBGS AEUUEIHPBRAIU RENTEN ET nmol VwZMDC S BDAMSIEOUENKMETISOBI I NOacH S CUNLES EHI NSIEREOSmIEFCHHBBIZhnhLEJ ADIRE EZE ERP DEVENII ET 410 nm RI L EXKREZWMELET VwUMOeEm
138. EDEBSE P 225C GE 410 nm uM UL nmol 225C GE 0 C 6 2 nmol 4 1 nmol 12 5 nmol 15 9 nmol 37 C 50 nmol 38 9 nmol JJ E E EX bos Te 1H gt H D LI Ta P 20 3 pH pH 5 pH pHS pH nmol E RP TET pH 63 pae 8 9 410 nm P nmol pH pH 63 piR6 90 909 pHS 0 12 5 nmol 17 4 nmol pH6 3 6 2 nmol 4 1 nmol pH8 6 0 4 nmol pH al E m OO N NN E 4 ps x t 1
139. H 2 H D LI A P 21 4 NM TERA X RIR BE FRAOICH ODN RAEC CTTO mic Oe Ei Db HER ET TORRT F EORR IY P nme 100 100 m 2 N H D LI a ts 1H RE 50 nmol 49 7 nmol 25 nmol 26 7 nmol 27 5 l
140. L S2 10mL 10 mL 5 12 1 5 mM 6 1 5 1 1 ea a 44066 UN DPTP 15 mL
141. Skinner Toothpickase http www accessexcellence org AE ATG data released 0166 PeggySkinner S 1 5 mM Lr a do dam 1 buie IER 1 1 S1 S5 1 DPTP 4 153b Proceso Js 6 Ea ur DPTP AIMO SIE c aT sues UU EX v 1 15 mL 1 DPTP 1 mL DPTP 1 mL DPTP TTTTTTTT fff E V U J 63 2 3 HI H3
142. mol CN 12 5 nmol 0 nmol 2 0 6 nmo1 9 nmol 0 nmol 2 0 4 5 nmol pH 5 0 nmol pH 6 3 nmol pH 8 6 nmol 61 2 pH R E E H pin Tes 1H 2 N H D LI L p 3 1 pH 2 p 3 pH pH 4 62 4
143. p 1 410 nm E p
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