たばこ乾燥葉の保存過程における 糖質成分の変化および
Contents
1.2. 1
3. 2 106
4. 10 12w
5. 122
6. 103 lt NE
7. 109 ERRE 2006 TASC 2009 FST BBB
8. 3 2 LC MS MS
9. RE EHRE 2006
10. R amp D ADE ATR R amp D GH R amp D
11. or w BB Anselme Payen Jean Francois Persoz Vandermaarel M J E C et al 2002 o a amylase 1 4 0 D glucan glucanohydrolase EC 3 2 1 1 endo o 1 4 p
12. 3 3 2009
13. 4 Lice
14. Dry basis D B 2 2 6 22 C 60 RH 48 40 C 60 RH 4 23 2 3 2 3 1 2 1
15. me Maharaj V and Sankat C K 1996 Tom s Barber n F A and Esp n J C 2001 Moreno F J et al 2006 FE Negi P S and Roy S K 2001 Linbo S et al 2007
16. mvertase B D fructofuranosidase EC 3 2 1 26 Sucrase sucrose a D glucohydrolase EC 3 2 1 48 Cairns A J and Ashton J E 1991 Ende W V D and Laere A V 1993 Obenland D M et al 1993 Agopian R G D et al 2009 4
17. drying curing Lamina L Curing _ Terrill T 1974 Tso T C 1999 1 2 aging gt ME RA FRAME 72 EDRR RML Er Z lt DOI o Sr
18. 1998 2007 1912 L C Maillard Maillard Maillard L C 1912
19. 3 2 Lk LC ESI MS MS
20. Young J R 11 and Jeffrey R N 1943 DREI b EST SIR FH Abubakar Y et al 2000 Curing Pucher G W and Vickery H B 1949 Vickert H B and Abrahams M D 1949
21. MS MS 3 3 2 LC ESI MS MS E ESD
22. 3 Sample Sample 1 1wt Sample 2 Swt Sample 3 Sample 2 120 C 10 Sample 2 3 5 6wt Sample 3 120 10 22 60 RH 24 3 40 C
23. 4 Ltr Lotto TORR amp Hart HORRBEPICILA FEARS KE lt Sie 3 2 B pH 2
24. op 5 LO RI ee BE 5 1 C OD 99 pou oui seupjox u 201 DLA poyyuenb sem soonjy y qnd 3AInQ UOUBIOUE qnd 3AImo uonzg iqI 8O LANZ AINO UOI1EIdIIE HaQVN Mb Joog oquieT te no o our UOHeoHHUen wu 0r dNd wu COP dNd WU 00 dNO SU DEE Soon UJSu o APA uono 1 p yonpol sgy SS p soonjo gJ q Nd p soon o V q Nd dNO g SD N 9SOIOnS ayeysqns uoneziyeyye BUI OOS 2 UOHBZIIBII uonezyeyye uoneziyeyye youond SI Ur 09 01 O0c S uru ST Sr Sr SS Le O S OV O S Oo dduo Hd HAG l CO k Se Aik AOE AOE AOE Jo m MW sep soonj3 g sep soon 3 p oseyAure 0 ISVLIOAU AiAnoV Su zuqg I s 100 A Invertase BRA FCV JPN USA BRA BLY JPN GRE ORI nkat g DB B a amylase BRA FCV JPN USA BRA BLY JPN USA GRE ORI TUR nkat g DB 5 1
25. HPAEC PAD Tabata S and Dohi Y 1992 Campbell J M et al 1997 Hogarth A J C L et al 2000 HPAEC PAD MSD 37 Wang J et al 1999 Mauri P et al 2002 Liu Y et al 2005 Penna N et al 2009 MSD PAD MSD MALDI TOF MS FT_ICR MS
26. oc o B ov Curing
27. ee ga ll i A Se ap SS curing CS zz PR curing a men mm L nen mm a api A SE 1 1 10 STE A PL k ELIET Oom FERRA teil ko C K lt EA curing Flue curing 30C 70C
28. IVI 60 C IV2 IV3 55 C pH IV3 4 8 4 9 IV1 gt IV2 gt IV3 4 10 4 11 BAR IV3 3 IV
29. Bourne E J et al 1967 Stedman R L 1968 Siddiqui I R and Rosa N 1983 Oakley E T 1983 vob Troje Z et al 1997 Hall R A and Wooten J B 1998 Clarke M B et al 2006 Tang K et al 2007 Z French D 1955 Mizuno T and Kinpyo T 1957 Siddiqui I R and Rosa N 1983 3 HO
30. Hodge J E 1953 Silvan J M et al 2006 1990 Vanderhaegen B et al 2006 Maillard
31. Smaniotto A et al 2009 4 1 1 0 o Enzyme activity Co Protein conc 0 05 Enzyme Activity nkat mL e Protein conc mg mL 0 00 0 10 20 Charcoal w v 4 1 69 12 wy DEE
32. 2 3 12 mL pH pH 4 C 2 2 3 100 mM pHS 0 Mcllvaine 200 pL 120 yL 40 C 2 21 4 R m 0 5 M 80 pL 5 10 20 40 1 0 M 200 pL JNA pH zl 1 1 nmol
33. 30 C 6 000 xg 15 19 g 4g 80 mL Buffer D 4 800 xg 30 40 mL 30 kDa Amicon Ultra 2 FAV CHE MG Le 2 mL Buffer D 0 2 um lt Ju ZX 7 ET GE Healthcare UK Ltd 1 0 mL 4 2 6 y 4 2 8 pH 5 0 50 mM McIlvaine 100 mM 50 C 15
34. IS 3 Sturm A 1999 Wi SS pH _ CRW pH Fotopoulos V 2005 3 pH IVI IV2 4 3 2 IV1 1IV2
35. 1 DO 2 cy lt TY SO Kazmaier T et al 1998 Seipert R R et al 200 ZS 38 HO O O OH OH i Fructooligosaccharides FOS O I Inuline type G e n 0 DP3 1 Kestose n 1 DP4 Nistose n 2 DP5 Fructosylnistose OH OH OH HO IM HO O OH O O OH OH Maltooligosaccharides MOS n 1 DP3 Maltotriose n 2 DP4 Ma
36. 1 1 3 1 1 2 Tso T C 1999 1 1 2
37. 3 5 3 3 FOS DP3 MOS DP4 3 4
38. BOR CL MEL W lt 18 Z 2 SR Pete P DAT iA or GER 2 1 Fe 1 1 6 1 1 5 H GE 2 H l 85 C 1 5 KE LOW H
39. 2 4 L 5 17 2 2 3 2 2 6 22C 60 10 11
40. Pilon Smits E A H et al 1995 Sprenger N et al 1997 Curing 3 6 3 2 2 3 5 THEIL 3 5 B Friedmann M 1996 F
41. BC 2013 1 1 5 1 1 6 1 2 1 3 2 2 1 Fe Told TARR EDO AZ a ASS fi 268 c BETH PRAT 2 2 e 1 Zin 2 2 2 23 2 2 4 2 25 2 2 6 2 3 2 3 1 11 13 15 16 17 20 20 20 21 21 22 23 23 24 24 2 3 2 pH 2 3 3 2 4 2 5 3
42. BLY FCV ORI HARE 1 2 4 10 20 50 100 200 mM 29 S v Michaelis Menten 2 5 OF 2 1 Km Vmax 3 2 1 Km Vmax Leaf type Growing district Km Vmax mM nmol s mL FCV Braz
43. Gager F L et al 1971a b Baker R R 1999 13 ge Tomasik P et al 1989 Leffingwell J C 1999 AU DRED 0 20wt Flue curing Sun air curing Air curing
44. 2008 0 5 1 0 mm 2 2 2 4 2 2 1 kg 0 1 Q10 Triton X100 4 8 mM 10 2 mM 15 mM Mcllvaine pH 5 4 Buffer A 5 L 4 C Miracloth Merck Ltd 4 800 xg 30 3 5 L Crude extract 63 4 C 4 2 3 a Crude extract 3 5L
45. Sucrose fructan fructosyltransferase SFT Fructan fructan fructosyltransferase FFT EC 2 4 1 100 Sprenger N et al 1995 Vergauwen R et al 2003 Henry R J and Darbyshire B 1980 Shiomi N 2008 1 Sucrose sucrose 1 fructosyltransferase 1 SST SFT FFT SST 59 Pranznik W et al 1990 L scher M et al 2000 Ghazi L et al 2007
46. 1 R 4 3 IV1 90 1 13 3 10 D B gt 300 D B
47. 2 2 5 2 2 4 2 6 40 C 4 31 The Residual Ratio of Sucrose in Cured Tobacco Leaves during Storage ISucl Suc ow 120 100 80 60 40 20 0 0 10 20 R 0 8076 30 40 50 60 Enzyme Activity nkat g D B 2 6 nkag D B
48. 30 Nicotiana tabacum amp Nicotiana rustica 2 4 7
49. Pilon Smits E A H et al 1995 Sprenger N et al 1997 MOS q1 4 7 1 U JE 2 Stedman R L 1968 Siddiqui I R and Rosa N 1983 vob Troje Z et al 1997 2 Abbott L R and Matheson N K 1972 Curing 3 1
50. 17 4 3 2 5
51. 3 2 4 Instrument Agilent 1200 HPLC 6410 triple quadrupole MSD Agilent Technology HPLC conditions 41 Column Develosil RP Aqueous Column 150 X 1 5 mm I D Nomura Chemical Co Ltd Gradient elution Mobile phase A 5 mM ammonium acetate Mobile phase B Acetonitrile A100 0 15 min A85 15 30 min linear gradient A85 30 40 min A0 20 min post run Flow rate 0 1 mL min Column temperature 30 C Injection volume 5 uL Electrospray ionization conditions e Negative ion mode Capillary voltage 4000 V Drying gas temperature 350 C Gas flow 11 L min e Nebulizer pressure 35 psi Multiple reaction monitoring MRM i MS MS 3 1 Product ion chromatography PIC MRM Injection volume 2 10 uL e Collision energy 5 35 V Scan time 5
52. IVI TIV2 IV3 IV2 IV3 4 2 RIC ZL h ZZ Z LY VBR E Fe WE EN E IV1 LG 2 4 4 2 4 5 100 mM IV1 PIC MRM 1 76 3 MIC CHR FOS DP4
53. 300 000 590 000 cDNA 3
54. pH 5 0 55 C 3 26 Glc production oc Fru production 100 80 60 40 Relative velocity 20 0 2 3 pH 40 C O Mcllvaine pH 100 0 08 M 40 C 40 27 e Glc productio
55. 15 mM Mcllvame buffer pH 5 4 Aik FBR IB ZO m OX KIRE 0 1 g mL 2 1 EP VERO FR AEA 24 80 mM 40 C 2 2 A B A HPLC RID chromatogram nRIU 1250001 f Sucrose 1 100000 Ak Kess Glucose 50000 Fructose 120 min 25000 e in min 0 Urn AN a 5 6 7 8 9 10 11 12 13 14 mm B Fructose Glucose Sucrose 20 20 90 15 15 80 10 10 S S e y 0 137x 1 957 lt y 0143x 2 099 70 R2 0 996 e R2 0 995 y 0 129 x 86 038 R2 0 977 0 0 60 0
56. Leffingwell J C 1999 Baker R R et al 2004a b Reinskje T et al 2006 eS Kee Glucose Sucrose OH H o SS O HO HO OH OH Maltose Fructose 1 2 1 1 6 1 3 50 40 8 A e
57. mvert Invert Lee H S and Sturm A 1996 Ishimoto M and Nakamura A 1997 Belcarz A et al 2002 Nakamura Nakamura M et al 1988 Greiner cDNA
58. 60 4 4 1 P 3 SST B D fructofuranosidase E C 3 2 1 26 Invert sugar
59. HILIC H ix C30 C30 44 100 DAHA AAV S SEDS AYRE Ca 0 Ee GE OA THe C30 MS MS
60. c 94 tia CURSE DIC R I BORER n B CER gt FXO BRR a CHO FYI VRBO mv HORST Bean As lt 78 DIBA c b 2 7 Curing oc B 4 E SUR QO Bi pane rH IT e
61. 1 1 4 1 1 3 1 1 6 16 SEN EES EE ek LI Ke Sh oN bel H 1 1 1 1 1 2 Y 1 3 1 6 2
62. Sheen S J and Calvert J 1969 Weston T J 1968 Wahlberg L et al 1977 Curing Air curing Flue curing Curins aging PS Terrill T 1974 Tso T C 1999
63. pH Km pH WARE Mcllvaine RE T 4 2 9 MRM 3 2 4 PIC Ht LC PIC MRM 3 2 4 HPLC conditions Column Develosil RP Aqueous Column 150x1 5 mm I D Nomura Chemical Co Ltd e Gradient elution 5 mM ammonium acetate isocratic elution Flow rate 0 1 mL min Column temperature 30 C Injection volume 2 uL 67 PIC conditions Collision energy 10 V S
64. aging decarboxylase Sun curing PPO PAL a Sun J et al 2010 1 1 5 Weeks W W 1999
65. PIC MRM A PIC B 30 PIC O D m z 503 MRM 78 4 4 3 4 4 4 F pH AAR IVI 60 C 4 6 IV2 TIV3 EE 55 C 4 7 pH IV1 pH 5 0 pH W pH 6 0 pH 4 8 IV2 IV3 EN pH 3 0 4 0 TIV2 IV1 amp Bi pH IV3 4 9 IV1 IV2 IV3 pH 4
66. 108
67. Curing 91 Z 5 5 1 Fe ZE RE CERM EER TI UAV AEDS EE TEP O AZ u 2 SHIT SEIN re 4 Curing Threshing Aging 10 12
68. D D 1 BAL I Yea Oy eka RY I LCMS HPLC 18 2 MQ cm MilliQ Millipore Co EEN DPRIERR 22 C 60 RH 48 7 C 22 C 30 C 40 C 4 12 3 2 3 20 o MiniBlender Melitta 40 Japan Ltd 1 2mm 1 000 0 001 g 0 mL 40 mL 50 30 200 rpm 5
69. B 2 4 CNP pH Be CNP oc qa 60 97 1 1 nmol CNP 1nkat 5 2 6 oe 10mM pH 5 5 1 0 mL 100 yL 2 45 C 2 20 mM 4 o D Glc a pNP 00 pL
70. o oc 2 1 2 1 1 2
71. 3 1 JF 3 2 3 2 1 3 2 2 3 2 3 3 2 4 3 3 3 3 1 3 3 2 LC ESI MS MS 3 4 3 5 GE 3 6 3 7 3 8 4 4 1 P 4 2 26 29 31 33 36 40 40 40 40 41 44 44 45 48 48 52 54 59 62 63 4 2 8 4 2 9 4 2 10 P 2 11
72. Frankenburg W G 1950B Frankenburg W G 1950B 12 Prabhu S and Chakraborty M 1986 Dixon Dixon L F et al 1936 Barret Barrett R E 1957 aging aging
73. 4 3 4 3 1 Bee 4 4 4 4 1 4 4 2 4 4 3 4 4 4 4 4 5 4 4 6 pH 4 5 4 6 63 63 64 64 65 65 65 66 67 68 68 69 69 73 76 76 76 79 79 79 85 87 89 5 1 P 5 2 5 2 1 5 2 2 GL ar 5 2 3 5 2 4 5 2 5 o 5 2 6 wc 5 2 7 BB 5 3 5 4 ei d I 6 Si DI 93 95 95 96 97 97 97 98 98 99 103 104 110 121 s N SE 1 1 AAO A se 1 1 1
74. IV1 OH OH o HO OH O O OH OH Sucrose H2O Hydrolysis Fructose Sucrose Transfructosylation Kestose Glucose 1 kestose amp Isomers 4 12 IV1 1 4 13 1 1 87 A glucose 1 kestose v nmol st mL 0 100 200 300 Sucrose mmol L foe a B The ratio of 1 kestose formation rate to glucose formation rate 0 100 200 300 Sucrose mmol L 4 13 IVI A
75. 3 6 4 Sample 1 2 Sample PIC PIC MS MS 3 2 PIC 54 3 7 Sample 2 Sample 3 Sample 2
76. Feu N tabacum BY 2 cells 70 000 Nakamura M et al 1988 cDNA 61 000 Greiner S et al 1995 IV3 70 000 IV1 IV2 4 8 IVI Dithiothreitol SDS PAGE Blue native PAGE BN PAGE 4 4 CBB PAS
77. 1 nkat 2 2 4 2 1 2 2 3 1 0 M F kit Glucose FE kit Glucose Fructose F kit 2 2 2 3 1 0 M 832 pL 10 kDa Amicon Ultra 4 mL Agilent EIEE ax Agilent 1200 HPLC system G1362A refractive index detector HPLC RID 0 1 10 mgmL
78. 4 11 nkat mL 1 6 1 4 Le 1 0 0 8 0 6 0 4 0 2 0 0 80 C Mclivaine f 50 C 35 C 60 C 65 C W 6 gt 80 C 1000 2000 Heating Time s 4 10 IV1 50 C 55 C 60 C 65 C 70 C HUX pH 5 0 300 1500 4 2 8 85 Le A 1 4 12 50 C 1 0 35 C Z 0 8 60 C X 0 6 65 C 0 4 370 C w 80 C 0 0 0 1000 2000 Heating Time s 1 6 B 1 4 1 2 50 C 1 0 55C 0 8 60 C 0 6 65 C 0 4 IFC 2 80 C 0 0 0 1000 2000 Heating Time s 4 11 IV2 IV3 A IV2 B IV3 50 C 55 C 60 C 65 C 70 C 80 C Mcllvaine S pH 5 0 300 1500 4 2 8 86 4 5 IV1 1 RA 12
79. A B c FCV Flue cured Virginia BLY Burley ORI Oriental leaf USA BRA JPN HAZE TUR Lag GRE 101 C o glucosidase FCV BLY ORI nkat g DB D B glucosidase BRA l FCV JPN USA BRA i 7eo BLY JPN E 4310 USA Gz 96 0 GRE 7 5 ORI TUR 3 7 0 100 200 300 400 500 nkat g DB 5 1 O oc D B V FCV Flue cured Virginia BLY Burley F ORI Oriental leaf USA XEHE BRA JPN TUR GRE 102 5 4 ou o B
80. Greiner S et al 1995 62 Nakanishi K and Yokotsuka K 1990 Curing Threshing Aging 3 4 2 4 2 1
81. 1 oe anomer B anomer 1995 o 1995 c co 1 4 wc B B glucosidase B D glucoside glucohydrolase EC 3 2 1 21 B
82. ES HPLC 18 2 MQ cm MilliQ Millipore Co Bedford MA USA 2 2 2 Nara Machinery Co Ltd 0 5 1 0 mm 2 0 gs 100 mL 15mM Mcllvaine 4 8 mM 10 2 mM pH 5 4 30 Whatmann 60 12 000 xg 10 02 um Whatmann GE Healthcare UK Ltd 60 mL 30 kDa Amicon Ulra x4
83. 2 G r SEI TEA pH pH 5 0 HEI 50 00 105
84. 2 3 pH5 0 40 C 2 3 34 3 3 1 IT IT el TE e OS O G U EK TT S Wa O DM Leffingwell J C 1999 Weeks W W 1999 Baker R R et al 2004 Reinskje T et al 2006
85. 5 2 3 5 1 pH w REDS 55 C 40 C 5 2 4 2 2 2 3 5 2 5 w wc e 2 4 6 6 B N3 GS B CNP o amylase
86. IV2 IV3 4 2 IV1 Substrate Activity nkat mL Relative activity Fru Gle Sucrose 1 21 90 1 35 100 Raffinose 0 98 73 N D Stachyose 1 03 76 N D 1 Kesotse 0 97 72 trace Melezitose N D ND Maltose N D Trehalose trace a 100 mM substrates were used in each test The rate of glucose formation using sucrose as a substrate is defined as 100 Fructose and glucose were quantified using F kit 2 N D means less than 0 03 nkat mL trace means less than 0 07 nkat mL Substrates Structure Sucrose Glc o1 lt 2B Fru Raffnose Gal a1 6 Glc a1 lt gt 2B Fru Stachyose Gal a1 6 Gal a1 6 Glc a1 2B Fru 1 Kestose Gle a1 lt gt 28 Fru B1 lt gt 26 Fru Melezitose Glce a1 3 Fru 62 lt 1a Gle Maltose Gle al 4 Glce Trehalose Gle al lt gt la Gle 77 abundance abundance 1 estose A PIC nistose Standard chemicals me AA EE B PIC Enzymatic reaction product 30 min e LA 1 5 min Ge os MRM W 10 min NV rin m z 503 323 4 AN 30 min D MRM m z 503 221 2 4 6 8 10 12 4 6 18 26 Retention time min 4 5 100mM IVI
87. 4 B_D 5 2 2 Nara Machinery Co Ltd 0 5 1 0 mm 4 C 2 0 g 100 mL McIlvaine 15 mM 4 8 mM 10 2 mM pH 5 4 30 Whatmann 60 12 000 xg 10 0 2 um Whatmann GE Healthcare UK Ltd oc 0 02 e 1 mL 30 mL 30 kD
88. 30 10 kDa Amicon Ultra 15 mLX2 20 mL P RU 5 mL MiliQ 10 mL 0 2 um PVDF HAWG EOS HEE HPLC RID 2 2 2 4 Product ion chromatography PIC _ CE 1 mL Oasis MCX Cartridge 6 cm7150 mg Waters Co 2 mL 0 2 um PVDF
89. h Huy CHEERS Q sepharose Fast Flow Concanavalin A sepharose 4B 4 1 V Al GFC 4 2 GFC 4 2 A GFC IV1 IV2 4 2 B IV1 IV2 GFC
90. W 2 Troller J A and Christian J H B 1981 ay 0 9 ay 0 8 g 0 7 10 12 0 6 1 2 He CH O 1 1 5
91. P amylase 1 4 c D glucan glucanohydrolase EC 3 2 1 2 exo m Ka TEV MARIA wam LC HLL CHAK yfi L et F AREER SE E Glucoamylase 1 4 a D glucan glucohydolase EC 3 2 1 3 exo 93 exo 4 1 4 c D glucan maltotetraohydrolase EC 3 2 1 60 6 1 4 c D glucan maltohexaohydrolase EC 3 2 1 98 Robyt J F and Ackerman R J 1971 Momma M 2000 lsoamylase glycogen a 1 6 glucanohydrolase EC 3 2 1 68 wc o glucosidase o D glucoside glucohydrolase EC 3 2 1 20 a 1 4 w 1 4 7 EC 3 2 1 3
92. 80 120 1 FOS DP3 1 0 50 ug mL 0 05 5 0ug mL 3 3 2 1 2 Air curing HUGS REI Curing Leffinsgwell J C 1999 48 A All monitored ions of MRM B m z 503 gt 323 C m z 665 485 D m z 827 gt 647 abundance
93. 5 7 Peedin G F 1999 Abubakar Y et al 2000 Air curing 30 40 Palmer G K and Pearce R C 1999 Sun air curing PEMA 20 40C 60 70 RH Gilchrist S N 1999 1 1 4 Curing 1930 1950 Frankenburg Frankenburg W G 1946A Curing
94. 1 B 1 88 46 i IV1 IV2 IV3 3 1IV1 1IV2 2 B pH pH
95. 3 TO lEs i IV1 IV2 IV3 Crude extract IV1 414 fold IV2 63 fold IV3 28 fold Crud extract GFC 4 1 eye AO BS HHI 4 2 CO GFC 4 2 7 LU RS IV1 1IV2 70 wi e Invertase Activity UV absorbance wu 087 39uedIOSdV AN Co e e CIE AWANOY SSBJIGAUI uru 08 lt soueqlosqV AN 5 en en Q Se 6S Fae 80 90 100 Volume mL E 70 60 Elution 50 TueYU AJIAHOV 3SejJ9AU 60 70 80 90 100 Elution Volume mL 50 40 wu 087
96. Waters Carbohydrate Column 250x4 6 mm I D 4 um 35 C BHA 75 1 0 mLmin 20 uL HPLC RID 22 1 0 g 50 40 mL 30 200 rpm 30 02 um PVDF Whatmann GE Healthcare UK Ltd HPLC RID Wet basis W B 2 2 5 1 0 g 100 C Tsukasa Co Ltd Tokyo 1
97. 1 M pH 10 4 C 30 kDa Amicon Ultra micro centrifugal tube F kit Glucose F kit Glucose Fructose BR F Roche Applied Science Switzerland 66 1 1 Agilent 1200 HPLC 6410 Triple Quadrupole MSD LC MS MS Agilent Technology
98. 4 3 7 A 2 Sample 3 3 7 B Sample 3 3 8 Sample 2 Sample 3 Sample 2 3 8 A 1 DP3 DP4 DP5 1 2 4
99. IVI GFC IVI IV2 IV1 IV2 73 o wn INI 590k 13 TV2 300k IV3 70k 21 A 11 O Molecular mass markers 1Vs 10 0 0 1 0 2 0 3 0 4 0 5 0 6 Kav 4 3 Kav Ve Vo Vc Vo Ve elution volume Vc column volume Vo void volume Void volume Vo was determined with blue dextrin Mr relative molecular mass Molecular mass markers Thyroglobulin 669k Ferritin 440k Aldolase 158k Conalbumin 75k Ovalbumin 44k 74 Pre stained A B C Standard
100. Suclow Suclaw 40 C 4 17 32 2 5 pH pH 5 0 50C
101. a A 40 52 ppm D B ppm D B 1600 1400 1200 1000 800 600 400 200 300 250 200 150 100 50 3 5 A Fructooligosaccharides m FOS DPS FOS DP4 E FOS DP3 CTL before storage TE 22 C 30 C 40 C after 12 weeks storage at each temperature B Maltooligosaccharides CIL before storage PC mMOSDP7 MOSDP6 MOSDPS mMOS DP4 B n 22 C 30 C 40 C after 12 weeks storage at each temperature A B 53 3 7
102. 1990 45 403 410 2007 MHA 45 177 185 1998 Nippon Nogeikagaku Kaishi 69 1050 1054 1995 Abbott I R and Matheson N K Starch depletion in germinating wheat wrinkled seeded peas and senescing tobacco leaves Phytochemistry 11 1261 1272 1972 Abubakar Y Young J H Johnson W H and Weeks W W Changes in moisture and chemical composition of flue cured tobacco during curing Tob Sci 44 51 58 2000 Agopian R G D Purgatto E Cordenunsi B R and Lajolo F M Synthesis of fructooligosaccharides in banana Prata ana its relation to invertase activity and sucrose accumulation J Agric Food Chem 57 10765 10771 2009 Bahr U Pfenninger A and Karas M High sensitivity analysis of neutral underivatized oligosaccharides by nanoelectrospray mass spectrometry Anal Chem 69 4530 4535 1997 110 Baker R R Smoke chemistry p 398 439 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Baker R R Pereira da Silva J R and Smith G The effect of tobacco in
103. Bahr U et al 1997 Liu Y et al 2005 Penna N et al 2009 SIN M H 45 MOS DP3 lt MOS DP7 m z 989 MS MS
104. Sample 2 1 TT Sample 2 Sample 3 3 8 B 3 7 B 55 EUKI O EA SW 4 4 SH 4 EO d WOT Old ULLAN HY Bt 0 0p Ee MLA LUTE A BDO 0p EY FH 1 OH 3IdmeS Vu Zi nk Pepues 4 4 SA Old kaler Ellen O ER Z UU Z UU Z UU 008 00L 009 00 00L 009 00 00t 00 00 00t 00E 007 001 ID A 0 6 OTHE CC t OTE Lr9 0 CE O 6LI 0 8r CEOS TETE T LTS 99 q y uw own uonus1o OV SE oF SC OC S oO sS
105. amp 107 1 5 CRRA OL OREO w w B
106. as F eASOIINS MOUM I atdhge i V 3SOIOQS Ota 3 Z i 3 q Z z aldweg Vv piepue1s Pa 3soJSruIAsolomrd cd 3sonsIN dQ Ssols 3 I cd ouepunqe SAHEISS AJISUS1UI 56 A x10 Sample 2 with sucrose untreated 70 60 Fructose 50 Glucose 4 A 40 Sucrose 30 20 10 0 0 2 4 6 8 storage period week B x103 Sample 3 with sucrose heat treated Fructose Glucose Sucrose srorage period week 3 7 1 A Sample 2 B Sample 3 57 A 900 800 700 _ 600 500 400 300 200 100 Sample 2 with sucrose untreated storage period week Sample 3 with sucrose heat treated 0 2 4 6 8 storage period week 3 8 E LEI e rH OHS oR AE 2 DP 3 5 A Sample 2 B Sample 3 5
107. 0 1 Triton X 100 300 mM NaCl pH 6 0 20 mM Bis Tris 200 mL mL min 30 kDa Amicon Ultra Buffer C 20 mM Bis Tris 500 mM NaCl 1mM MnCl 1mM CaCb pH 6 0 50 mL 64 4 2 5 4 2 4 50 mL Concanavalin A Sepharose 4B Fast Flow Column GE Healthcare UK Ltd 5 mL 2 Buffer C 50 mL Buffer C 500 mM NaCl 1 M ov D 20 mM Bis Tris WYK 100 mL MOTRA LE 1 mL min OM WIRE 30 kDa Amicon Ultra Buffer D 20 mM Bis Tris 100 mM NaCl p
108. tt tt 2 ad a i f tf id a t amp tf ot ft a amp tt ud i o ce nh ao F m z 827 665 Retention Time min 3 3 Kelte A B FOS DP3 O FOS FP4 D FOS DP5 E MOS DP4 F MOS DPS G MOS DP6 DP7 49 Tu 473 Tse Ot 0 00 O 0 000S 00001 ooost 00007 Z 000 Sz S Li 0000 000 lt c 0000 89666 0 z4 Ladd SOW Tw 4 3 1028 SO 0 To 8666 0 7a Sd SO SEA BE EK HAEL Dr Eco 9OURPUNQW Bi Cy DISA nt O YY BAIA b ELI Tum Ar TNs Tuom 4 AH Twm A Ts Or OZ 00 OF Or OZ 00 OF ct o lt 0 c 0 0 0 SS p F 000c 8666 0 000c 8666 0 8666 0 000c A 00001 Weder ooost 00001 Z 000ST Z 00007 amp 8 00007 8 OS 000SI 0000c 0005 000s 00007 0000 0000r 9dd SOW Sda SOW tdd SOW mom Ke mom Ar A 0 OE OT OT OO r et OE ST 0 ST OE 7 0 0 d 7 S 00001 oer 8666 0 ost 00007 gt 007 amp 0000 amp osz 8 8 0000F oe a 0000c om 00009 tda SOA dd SO 50 Joyreu asoueder ye paseyoind alam g pue v adures Sa1SIB8TO AIIeIOISUTUTOO woy Mo uye 319A 19npoid ay Jo JOTI MO sam Daeadar aaA 19BXa ooogqo1 Jo UoHeIedaid Surpni ur sts euv popou pue penau
109. 15 50 mM 400 uL 50 mM 400 pL 98 Glc B pNP 500 pL Glc B pNP pH 9 5 Rm OMERE yor pNP 405 nm 1 1 nmol 4 pNP Inkat 5 3 5 2 8 5 1 A
110. 3 2 MS 162 amu FAB Garozzo D TH etal 1990 Collision Energy 5 30 V 180 amu 18 30 60 90 amu Garozzo D et al 1990 Bahr U et al 1997 180 amu 162 amu 180 amu KAY SHELL hAY SH
111. 12 w v Sigma Aldrich Co 3 NEAL lt FLE TOR W Buffer A 0 445 um NALGENE Nalge Nunc International Co 50 mL 30 kDa Millipore Co WETA 0 1 Triton X 20mM Bis Tris pH 6 0 Buffer B 1L 50 mL Buffer B 50 mL 4 2 4 50 mL Buffer B Q sepharose fast flow Column volume 100 mL GE Healthcare UK Ltd 300 mL Buffer B
112. IV3 IV3 QGFC IV3 89 70 000 IV3 Nakamura 70 000 Nakamura M et al 1988 pH 4 0 55 000 70 000 Wiz GFC TIV3 DS V3 BE IV1 IV2 IV3
113. 30 50 mM 400 uL 50 mM 400 pL Glc o pNP 500 pL Glc a pNP pNP 405 nm 1 1 nmol 4 pNP 1nkat 5 2 7 B 10mM pH 5 5 1 0 mL 100 pL 2 45 C 2 20 mM 4 o D Gilc B pNP 500 uL
114. FOS B 3 Ritsema T and Smeekens S 2003 Biggs D R and Hancock K R 2001 Banguela A and Hern ndez L 2006 36 15 Hendry G A F 1993 B2 1 B2 6 B2 1 B2 6 Hendry G A F 1993 Ritsema T and Smeekens S 2003
115. and Hunter D E Selected fructooligosaccharide I kestose nystose and 1 fructofuranosylnystose composition of foods and feeds J Agric Food Chem 45 3076 3082 1997 Clarke M B Bezabeh D Z and Howard C T Determination of carbohydrates in tobacco products by liquid chromatography Mass Spectrometry Mass Spectrometry A comparison with ion chromatography and application to product discrimination J Agric Food Chem 54 1975 1981 2006 Dixon L F Darkis F R Wolf F A Hall J A Jones E P and Gross P M Flue cured tobacco natural aging of flue cured cigarette tobaccos Ind Eng Chem 28 180 189 1936 Ende W V D and Laere A V Purification and properties of an invertase with surose sucrose fructosyltransferase SST activity from the roots of Cichorium intybus L New Phytol 12 31 37 1993 Fotopoulos V Plant invertases structure function and regulation of a diverse enzyme family J Biol Res 4 127 137 2005 Frankenburg W G Chemical Changes in the Harvested Tobacco Leaf I Chemical and Enzymatic Conversations during the Curing Process Advances in Enzymolozy Volume VI 6 309 387 1946 Frankenburg W G Chemical Changes in the Harvested Tobacco Leaf II Chemical and Enzymatic Conversations during Fermentation and Aging Advances in Enzymolozy Volume X 6 325 434 1950 French D Isolation and identification of planteose from tobacco seeds J Am C
116. 250 000 SA 150 000 75 000 50 000 4 4 IV1 A 3 ug CBB 4 B 15 ug CBB C 3ug PAS 75 4 4 IV1 4 4 1 IV1 4 2 HEE EH pH 100 IV1 1 70 IV1
117. 30 60 90 120 150 0 30 60 90 120 150 0 30 60 90 120 150 min min min 2 2 A HPLC RID B 25 2 2 2 2 3 2 pH 2 3 pH 2 4 pH 5 0
118. 4 5 EEIT OR Sturm A 1999 Pyridoxal Pressey R 1968 Lopez M E et al 1988 IV1 Pyridoxal DYS 4 3 79 nkat mL nkat mL 2 0 1 5 1 0 0 5 0 0 0 20 0 15 A 10 20 30 40 50 60 70 80 90 Temperature C B 10 20 30 40 50 60 70 80 90 Temperature C 4 6 IVI 25 C 70 C A B 1 A L nkat mL U 10 20 30 40 50 60 70 80 90 Temperature C 2 B A nkat mL an 10 20 30 40 50 60 70 80 90 Temperature C
119. l pH 4 5 5 0 DIRENIK H mj SLL HILL BERG Ae 10 12 0 60 0 65 2 2 2 2 1 D D 12 PED IRV COC SE CDV YC uh 17 20 2 EH UE HPLC
120. mM Pyridoxal 18 3 1 0 31 3 0 7 IV1 Pyridoxal IVI 84 4 4 6 McIlvaine LEAK Ge H TIV1 50 C 55 C 60 C 65 C 70 C 80 C pH 5 0 300 1800 4 2 8 4 10 IV1 55 C 1800 70 60 C IV2 IV3 IV2 IV3 IV1
121. soueqlosqV AN t U U y i lt CH keng TWU AJIAHOV SpLI9AU 50 60 70 80 90 100 40 GFC 4 2 A 1 load B re load ie A B 2 C ARN LC Reng B gt lt 71 DAD uo SUIpeol 21 Ad poyund sem TAI uongen genin s pn oui Puooss r d asoonys jowu Jo uonguttor a Surz g eo urKzu Jo JUNOW AJL soon 8 Jo uonguro oy SUIZATB1BO SB Data 8 lt T Fre SCH ZLI EAI 007 xepiedng 9 6 SIZ gr 61E TAI 007 xepsedng Fit CO Ort 0 0 LS IAI p 00 lt xepsedng rc II LIL 0I 0 I esoreydeg y no Ce oz STE ETL LITT esoreydag O ge cc gei el gege gt JUULIL E02IEYD payeanoy ere SCC SLITI pexa apni Piot mralord men 3u e qu uonpgourmd PISA a noe HITOadS U3 01d s AJIAHSY da uongourmdq SR E Uy k CO liese EI I r2Z 72 4 3 2 GFC IV1 Mr 590 k IV2 IV3 300k 70k 4 3 50 000 70 000 Fotopoulos V 2005
122. 00 1000 ms 42 auy uonu 1 1 LY ABJamra voten FD uonezu u od Jo m p Gd 9 T O ZI oor LTS DI 686 OFT L SOI TIls6 oor CS Ol 686 OFT 9 SON 9 8 0 L oor 99 DI LTS OFT SON cH oor I9I DI 99 OFT F SOI E0 FIE oor Leo Of LTS 091 04 ctl oor S8F SC 99 091 t 04 9 os ETE DC GI OFT SOA umu Ly sw opt matt zw urppoad A T9 Zrmm norosmoax LA onmamdgr dq pmodmo E X Cy COSIVSIN ARRO AS de I E 43 3 3 3 3 1 MSD
123. 30 o B go o C 20 e D e E 10 0 0 2 4 6 8 weeks 1 3 40C 60 6 8 15 1 3 ER N Maillard
124. 4 7 1V2 1V3 25 C 70 C A IV2 B IV3 1 6 1 4 1 2 1 0 0 8 0 6 0 4 Acetate buffer e McIlvaine buffer 0 2 Phosphate buffer 0 0 nkat mL 0 16 0 14 0 12 0 10 0 08 0 06 Zon Acetate buffer 0 02 O Mcllvaine buffer gt Phosphate buffer 0 00 4 8 IV1 pH pH 3 5 5 0 TIMERE pH 5 0 6 5 McIlvaine 87 pH 6 5 8 0 TIZ U Bo g EX amp AO A B 1 82 Acetate buffer Mcllvaine buffer Phosphate buffer A nkat mL Acetate buffer Mcllvaine buffer Phosphate buffer B nkat mL 4 9 IV2 IV3 pH pH 3 5 5 0 pH 5 0 6 5 Mcllvaine pH 6 5 8 0 A IV2 B IV3 83 4 3 IV1 Metal ions Glucose 1 Kestose Concentrations formation formation Inhibition Rate Inhibition Rate 200 mM MnCb 30 0 1 0 61 7 1 8 10 mM CuCls 60 6 0 4 53 8 1 0 10 mM ZnCl 29 4 1 2 34 7 0 2 10 uM HsCb 73 7 3 1 40 8 0 3 10
125. 8 3 8 LC ESLMS MS Sucrose sucrose fructosyltransferase 1 SST EC 3 2 1 99 Henry R J and Darbyshire B 1980 Pranznik W et al 1990
126. 912 Mauri P Minoggio M Simonetti P Gardana C and Pietta P Analysis of saccharides in beer samples by flow injection with electrospray mass spectrometry Rapid Commun Mass Spectrom 16 743 748 2002 Mizuno T and Kinpyo T Studies on the carbohydrates on tobacco Part I Free sugars and the constructive sugars of glycosides in green leaves of Japanese bright yellow tobacco J Agr Chem Soc Japan 31 297 299 1957 Momma M Cloning and sequencing of the maltohexaose producing amylase gene of 115 Klebsiella pneumonia Biosci Biotechnol Biochem 64 428 431 2000 Moreno F J Corzo Marti nez M Del Castillo M D and Villamiel M Changes in antioxidant activity of dehydrated onion and garlic during storage Food Res Int 39 891 897 2006 Nakamura M Hagimori M and Matsumoto T Purification and characterization of acid invertase from cultured tobacco cells Agric Biol Chem 52 3157 3158 1988 Nakanishi K and Yokotsuka K Characterization of thermostable invertase from wine grapes J Ferment Bioeng 69 16 22 1990 Negi P S and Roy S K Effect of drying conditions on quality of green leaves during long term storage Food Res Int 34 283 287 2001 Oakley E T Enzymatic determination of starch in fresh green lyophilized green and cured tobacco J Agric Food Chem 31 902 905 1983 Obenland D M Simmen U Boller T and Wiemken A Purif
127. CPMAS NMR J Agric Food Chem 46 1423 1427 1998 Hendry G A F Evolutionary origins and natural functions of fructans a 113 climatological biogeographic and mechanistic appraisal New Phytol 123 3 14 1993 Henry R J and Darbyshire B Sucrose sucrose fructosyltransferase and fructan fructan fructosyltransferase from allium cepa Phytochemistry 19 1017 1020 1980 Hodge J E Dehydrated foods chemistry of browning reactions in model systems J Agr Food Chem 1 928 943 1953 Hogarth A J C L Hunter D E Jacobs W A Garleb K A and Wolf B W Ion chromatographic determination of three fructooligosaccharide oligomers in prepared and preserved foods J Agric Food Chem 48 5326 5330 2000 Homma S Terasawa N Kubo T Yoneyama Ishii N Aida K and Fujimaki M Changes in chemical properties of melanoidin by oxidation and reduction Biosci Biotech Biochem 61 533 535 1997 Ishimoto M and Nakamura A Purification and properties of B fructofuranosidase from Clostridium perfringens Biosci Biotech Biochem 61 599 603 1997 Jay G D Culp D J and Jahnke M R Silver staining of extensively glycosylated proteins on sodium dodecyl sulfate polyacrylamide gels enhancement by carbohydrate binding dyes Anal Biochem 185 324 330 1990 Lee H S and Sturm A Purification and characterization of neutral and alkaline invertase from carrot Plant Physio
128. EL MOS DP4 7 FOS DP3 5 lt Colliision Energy 3 1 46 ALLS AGE LSD uonezuiou od Jo 99189q d SOW SEK SOA ECEIZA MN nk lee RON EAKL PTPN uor cAneSON SW SIA ISd 7 ELA zu zu zu zu 0001 006 008 00L 009 00S 000 006 008 00L 009 00S 006 008 00L 009 00S 00 ODE 009 OOS OOF OOF ONT OOT T SS IT lt PEOS TIPE S99 Z L738 6 09 Lda SOW 9d SOW cd SOW taq SON oO zm zm zm 008 00 009 00S 00L 009 00S OOF 00 oos OOF 00 00 00 2 Los OTHE cLt9 TSS 68LI A Ei TU eos Eze ZEN STSOH rd Sod ed SO 47 3 4 3 3 MRM
129. H 6 0 10 mL 4 2 6 4 2 5 30 kDa Amicon Ultra 500 pL Acta design FPLC Frac 950 fraction collector GE Healthcare UK Ltd Buffer D Superdex 200 HR16 60 gel filtration column GE Healthcare UK Ltd Buffer D 0 5 mL min Void volume 42 mL 7 2 0 mL 5 0 mL 4 2 7 N tabacum 4 C 330 g 700 mL Buffer A 65 Miracloth Merck Ltd
130. a Amicon Ultra E A IRE x2 2 3 6 mL 0 1 g 1 mL 30 mL 30 kDa Amicon Ultra E AIRS IRE x2 mM pH 5 2 3 5 mM BERBER pH 5 5 6 mL cu 96 B 0 1 sg 1 mL
131. can time 500 ms Scan range m z 100 700 MRM conditions RT Fragmentor Precursor Ion CE ProductIon Dwell Time min V m z V m z ms 0 10 140 503 10 221 200 0 10 140 503 20 323 200 10 25 140 665 20 485 200 RT retention time CE collision energy 4 2 10 Bradford Bradford M M 1976 BSA 4 2 11 PAGE 7 5 Blue Native PAGE BN PAGE Schagger H and Jagow G V 1991 CBB Periodic acid Schiff PAS Zacharius R M et al 1969 Jay G D et al 1990 68 43 4 3 1 Homma S et al 1997
132. e H and Wu F Establishment and application of enzymological models for predicting quality of flue cured tobacco J Food Agric Environ 8 326 331 2010 vob Troje Z Fr be Z and Perovi Analysis of selected alkaloids and sugars in tobacco extract J Chromatogr A 775 101 107 1997 Tabata S and Dohi Y An assay for oligo 1 4 1 4 glucantransferase activity in the glycogen debranching enzyme system by using HPLC with a pulsed amperometric detector Carbohydr Res 230 179 183 1992 Tang K Liang L Cai Y and Mou S Determination of sugars and alditols in tobacco with high performance anion exchange chromatography J Sep Sci 30 2160 2166 2007 Terrill T Influence of harvesting variables Rec Adv Tob Sci 62 85 92 1974 Tomasik P Palasinski M and Wiejak S The thermal decomposition of carbohydrates Part I The decomposition of mono di and oligo saccharides Adv Carbohydr Chem Biochem 47 203 278 1989 Tomas Barberan F A and Espin J C Phenolic compounds and related enzymes as determinants of quality in fruits and vegetables J Sci Food Agric 81 853 876 119 2001 Troller J A and Christian J H B 1981 Trufelli H and Cappiello A Study on the maltooligosaccharide composition mucilage samples collected along the northern Adriatic coast Carbohyd
133. gredients on smoke chemistry Part I Flavourings and additives Food Chem Toxicol 42S S3 37 2004a Baker R R Pereira da Silva J R and Smith G The effect of tobacco ingredients on smoke chemistry Part II Casing ingredients Food Chem Toxicol 42S S39 S52 2004b Banguela A and Hern ndez L Fructans from natural sources to transgenic plants Biotechnol Apl 23 202 210 2006 Barrett R E Tobacco Aging Enzymes of Bright and Burley Tobaccos J Agric Food Chem 5 220 224 1957 Belcarz A Ginalska G Lobarzewski J and Panel C The novel non glycosylated invertase from Candida utilis the properties and the conditions of production and purification Biochim Biophys Acta 1594 40 53 2002 Biggs D R and Hancock K R Fructan 2000 Trends Plant Sci 6 8 9 2001 Bourne E J Pridham J B and Worth H G J Pectic substances in cured and uncured tobacco Phytochemistry 6 423 431 1967 Bradford M M A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principal of protein dye binding Anal Biochem 72 248 254 1976 Cairns A J and Ashton J E The interpretation of in vitro measurements of fructosyl transferase activity an analysis of patterns of fructosyl transfer by fungal 111 invertase New Phytol 118 23 34 1991 Campbell J M Bauer L L Fahey G C Jr Hogarth A J C L Wolf B W
134. hem 112 Soc 77 1024 1025 1955 Friedmann M Food browning and its prevention an overview J Agric Food Chem 44 631 653 1996 Gager F L Nedlock J W and Martin W J Tobacco additives and cigarette smoke I Transfer of D glucose sucrose and their degradation products to the smoke Carbohydr Res 17 327 333 1971a Gager F L Nedlock J W and Martin W J Tobacco additives and cigarette smoke II Organic gas phase products from D glucose and sucrose Carbohydr Res 17 327 333 1971b Garozzo D Giuffrida M Impallomeni G Ballistreri A and Montaudo G Determination of linkage position and identification of the reducing end in linear oligosaccharides by negative ion fast atom bombardment mass spectrometry Anal Chem 62 279 286 1990 Ghazi I Fernandez Arrojo L Garcia Arellano H Ferrer M Ballesteros A and Plou F J Purification and kinetic characterization of a fructosyltransferase form Aspergillus aculeatus J Biotechnol 128 204 211 2007 Gilchrist S N Production practices Oriental tobacco p 154 163 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Greiner S Weil M Krausgrill S and Rausch T A tobacco cDNA coding for cell wall invertase Plant Physiol 108 825 826 1995 Hall R A and Wooten J B Quantitative analysis of cellulose in tobacco by Be
135. ication and characterization of three invertases from barley Hordeum vulgare L leaves Plant Physiol 101 1331 1339 1993 Palmer G K and Pearce R C Production practices Light air cured tobacco p 143 153 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Peedin G F Production practices Flue cured tobacco p 104 142 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Penna N Capellacci S Ricci F Giorgi M Penna A Famiglini G Pierini E Kazmaier T Roth S Zapp J Harding M and Kuhn R Quantitative analysis 116 of malto oligosaccharides by MALDI TOF mass spectrometry capillary electrophoresis and anion exchange chromatography Fresenius J Anal Chem 361 473 478 1998 Pilon Smits E A H Ebskamp M J M Paul M J Jenken M J W Weisbeek P J and Smeekens S C M Improved performance of transgenic fructan accumulating tobacco under drought stress Plant physiol 107 125 130 1995 Prabhu S and Chakraborty M Development of aroma bearing compounds and their precursors in flue cured tobacco during curing and post curing operations Tob Res 12 175 185 1986 Pranznik W Beck R H F and Spies T Isolation and characterization of sucrose sucrose 1 D fructosyltransferase tubers of helianthus tubero
136. il ave 7 29 1 58 sd 0 28 0 04 RSD 3 8 2 4 BLY Brazil ave 2 89 0 30 sd 0 05 0 00 RSD 1 7 0 5 ORI Greece ave 5 18 0 60 sd 0 41 0 00 RSD 7 8 0 0 X 2 5 CAR LZ K 2 C Michaelis Menten 3 Km 2 5 7 5 Vmax Vmax i Michaelis Menten R 2 5 2 1 100 mM 30 100 mM 1 1 nmol 1 nkat
137. k E sz L CH RETO WEEP CR 5 2 5 2 1 8 9 12 2 2 2 5 Dry basis D B D 4 2 4 2 12 3 95 4 Fu Z o D
138. l 112 1513 1533 1996 Leffingwell J C Leaf chemistry basic chemical constituents of tobacco leaf and differences among tobacco types p 265 284 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 114 Linbo S Torri L and Piergiovanni L Light induced changes in an aqueous beta carotene system stored under halogen and fluorescent lamps affected by two oxygen partial pressures J Agric Food Chem 55 5238 5245 2007 Liu Y Urgaonkar S Verkade J G and Armstrong D W Separation and characterization of underivatized oligosaccharides using liquid chromatography and liquid chromatography electrospray ionization mass spectrometry J Chromatogr A 1079 146 152 2005 Lopez M E Vattuone M A and Sampietro A R Partial purification and properties of invertase carica papaya fruits Phytochemistry 27 3077 3081 1988 Liischer M Hochstrasser U Boller T and Wiemken A Isolation of sucrose sucrose 1 fructosyltransferase 1 SST from barley Hordeum vulgare New Phytol 145 225 232 2000 Maharaj V and Sankat C K Quality changes in dehydrated dasheen leaves effects of blanching pre treatments and drying conditions Food Res Int 29 563 568 1996 Maillard L C Action des acides amin s sur les sucres formation des m lanoidines par vote m thodique C R Hebd S ances Acad Sci 154 66 68 1
139. ltotetraose OH HO O n 3 DPS Maltopentaose HO OH n 4 DP6 Maltohexaose OH O n 5 DP7 Maltoheptaose O HO OH OH O O HO OH OH 3 1 39 3 2 3 2 1 Green leaf cut filler 2 1 2 2 2
140. n o Fru production 100 80 60 40 Relative velocity 20 0 40 45 50 55 60 65 70 75 Temperature C 2 4 pH 5 0 100 0 08 M Mclivaine Z Sr pH 5 0 40 28 2 3 3 2 5 3 2 2 2 2 2 3 Kinetic curves v nmol ml s CH CO 0 6 0 4 0 2 0 0 0 50 100 150 200 S mM 2 5 S v plot
141. nship between leaf chemistry and organoleptic properties of tobacco smoke p 304 312 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Weston T J Biochemical characteristics of tobacco leaves during flue curing Phytochemistry 7 921 930 1968 Young J R and Jeffrey R N Changes in certain water soluble nitrogenous constituents of burley tobacco during storage Plant Plysiol 18 433 438 1943 Zacharius R M Zell T E Morrison J H and Woodlock J J Glycoprotein staining following electrophoresis on acrylamide gels Anal Biochem 30 148 152 1969 121 aer DIE Be CAB
142. r Res 344 120 126 2009 Tso T C Seed to smoke p 1 31 In Tobacco Production Chemistry and Technology Davis D L Nielsen M T Eds Blackwell Science Oxford U K 1999 Vanderhaegen B Neven H Varachtert H and Derdelinckx G The chemistry of beer aging a critical review Food Chem 95 357 381 2006 Vandermaarel M J E C Vanderveen B Uitdehaag J C M Leemhuis H and Dijkhuizen L Properties and applications of starch converting enzymes of the a amylase family J Biotechnol 94 137 155 2002 Vergauwen R Van Laere A and Van den Ende W Properties of Fructan Fructan 1 Fructosyltransferases from Chicory and Globe Thistle Two Asteracean Plants Storing Greatly Different Types of Inulin Plant Physiol 133 391 401 2003 Vickert H B and Abrahams M D The metabolism of the organic acids of tobacco leaves effect of culture of excised leaves in solutions of d isocitrate and acetate J Biol Chem 180 37 45 1949 Wahlberg I Kerstin K Austin D J Junker D Roeraade J Enzell C R and Johnson W H Effects of flue curing and ageing on the volatile neutral and acidic constituents of Virginia tobacco Phytochemistry 16 1217 1231 1977 Wang J Sporns P and Low N H Analysis of food oligosaccharides using MALDI MS quantification of fructooligosaccharides J Agric Food Chem 47 1549 1557 1999 120 Weeks W W Leaf chemistry relatio
143. sus L Agric Biol Chem 54 2429 2431 1990 Pressey R Inhibition of invertases by pyridoxal and its analogues Biochim Biophys Acta 159 414 416 1968 Pucher G W and Vickery H B The metabolism of the organic acids of tobacco leaves effect of culture of excised leaves in solutions of organic acid salts J Biol Chem 178 557 575 1949 Reinskje T Antoon O and Amsterdam J G C Sugars as tobacco ingredient Effects on mainstream smoke composition Food Chem Toxicol 44 1789 1798 2006 Robyt J F and Ackerman R J Isolation purification and characterization of a maltotetraose producing amylase from Pseudomonas stuzeri Arch Biochem Biophys 145 105 114 1971 Ritsema T and Smeekens S Fructans beneficial for plants and humans Curr Opin 117 Plant Biol 6 223 230 2003 Schagger H and Jagow G V Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form Anal Biochem 199 223 231 1991 Seipert R R Barboza M Ninonuevo M R LoGascio R G Mills D A Freeman S L German J B and Lebrilla C B Analysis and quantitation of fructooligosaccharides using matrix assusted laser desorption ionization fourier transform ion cyclotron resonance mass spectrometry Anal Chem 80 159 165 2008 Sheen S J and Calvert J Studies on polyphenol content activities and isozymes of polyphenol oxidase and pero
144. ut paquosap popou seq 14812A ATD ye palen pA3 sem wdd 8eraAY SIAL OOBdOQ1 UI SJUNOUIE IY 0 DJ13ATOO IIM 1BTXa UL Sa1ATU Jo SJUNOUVY I TonezrraA od Jo 2a18aq 51 99 6S 9 96 oE re 8t asl 9 0 96 0 cCOF LS 0F LSEF DES FOE 8g 0 wdd as Leg SS Sh See FI IT c9 aN POL 68 L udd s3eisAV dG SON CN EL 68 vs ot Le lt 6 OS 610 LUV z 0 SEKR SE LUOF ITOF E wdd as Frot cogs 9 8 67 ESTE 9 aN LO S II9 wdd s3ersAY 9dSON EL Ge T6 9 gI VII r9 S I asl I 0 STEF 910 73 0 or 0 SE 0 ETOF r0 0 wdd as pes SCC ELI 99 WK 80 aN SL E lg wdd s3ersAY Sdq SON S I LY es og rE TI 9 II L EI asl 910 0I0 IO I cris 60 0F 8L 0F r9 0 wdd as Ga I ZL FSI 6L II 799 IPL CN tL 9 ot udd s3eisAV tdq SON Ce DNA OS LTOF DES mdd qs op Iert CN CN SII 2 aN aN aN wdd s3ersAV Sqdq SO COI oE 9 CC OS 67 0 8S 0 690 L0 0 mdd qs Lt 60 61 aN aN 00 01 917 aN aoe aoe udd s3ersAV td SO os ge ET L 9 69 oE asl TS EF LS 91 ST S LVEF 8 96 0 udd qs 68 0L 8Z ON ON FR sss aN Ie Tr SEI gdd 38esAV d SOA dsIdureS vy atduge Weder ueder om 235 ZIG ven pg VSN shbormsrp Zumos apnodmo pH mo Jeg UaaID JUHO amg BIUISILA p mm2O 2n 4 JeaT By Take KOs fl ee CO
145. xidase during air curing in three tobacco types Plant Physiol 44 199 204 1969 Shiomi N Food biochemical study on fructans and related synthesis enzymes J Appl Glycosci 55 25 33 2008 Siddiqui I R and Rosa N Low molecular weight carbohydrates of tobacco Tob Sci 27 130 134 1983 Silvan J M Lagemaat J V D Olano A and Castillo M D D Analysis and biological properties of amino acid derivates formed by Maillard reaction in foods J Pharm Biomed Anal 4 1543 1551 2006 Smaniotto A Bertazzo B Comai S and Traldi P The role of peptides and proteins in melanoidin formation J Mass Spectrom 44 410 418 2009 Sprenger N Bortlik K Brandt A Boller T and Wiemken A Purification cloning and functional expression of sucrose fructan 6 fructosyltransferase a key enzyme of fructan synthesis in barley Proc Natl Acad Sci USA 92 11652 11656 1995 118 Sprenger N Schellenbaum L Dun K V Boller T and Wiemken A Fructan synthesis in transgenic tobacco and chicory plants expressing barley sucrose fructan 6 fructosyltransferase FEBS Lett 400 355 358 1997 Stedman R L The chemical composition of tobacco and tobacco smoke Chem Rev 68 153 207 1968 Sturm A Invertases Primary structures functions and roles in plant development and sucrose partitioning Plant Physiol 121 1 7 1999 Sun J Yan T Tu S He J Si H Xi
Download Pdf Manuals
Related Search