( CIP) /. :, ISBN X Q52-53 CIP (2003 ) ( 021 ) /

Transcription

1

2 ( CIP) /. :, ISBN X Q52-53 CIP (2003 ) ( 021 ) / ISBN X/ Q 4 : 50.00

3 1961 ( RN A) 5-5 -,,,, :, ;, ;, FDA,,, 18, 36,,, ;,,,,,,,,,

4 1 ( 3) 2 (11) 3 ATP (16) 4 (26) 5 (31) 6 (36) 7 (37) 8 -(41) 9 ATP (44) 10 (48) 11 (53) 12 (58) 13 (61) 14 (63) 15 D- (69) 16 (78) 17 C T P (83) 18 ATP (86) 1 (93) 2 (97) 3 ( 100) 4 ( 103) 5 D293 DN A 5 - ( 108) 6 ( 114) 7 ( 116) ( 120) 9 ( 124) 10 ( 126) 11 D903 ( 130) 1

5 13 ATP ( 133) ( 139) 14 ( 142) 15 ATP ( 147) ( 151) 17 ATP ( 157) 18 ( 161) 19 ( 167) ( 172) 21 ( 178) 22 ( 184) ( 192) 24 PN Pase ( 196) 25 ( 199) 26 ATP CO2 ( 202) ( 207) ( 212) 29 D- ( 219) 30 ( 224) FU R ( 228) 32 D- ( 233) 33 ( 238) 34 ( 243) ( 249) 36 QZN0209 ( 254) 2

6

7 ,,,,, : ( 1) ; (2 ) ; ( 3) ; ( 4) ; ( 5) ; ( 6) ; ( 7) ; ( 8 ) [ 1 ], L- ; 6- ; ; L- ; L-; - [ 2 ], (1a) [ 3 ] (1b) [ 4 ],, (),,,, ATP, (1c) [ 5 ],,,,, 1,,,,,, 3

8 1 * ( ) ( ) * () 1. (1 ) a.,,,,,,, ( 2 ), 3 [ 6 ] 4 [ 7 ], , 24, [ 8 ],, ( ), [ 9 ],,,,, 4

9 3 * IM: L- 1. D L b., 5,,,,,,,,,, [ 10 ] 6,, 5 6 5

10 7,,,,,,, (8 ) [ 12 ],, 7 2. (1 ) 8 Coulet [ 1 3 ] 0.4mm (9 ), [ 14 ] ( Streptom yces S P.) (10 ), Vieth [ 15 ] ( 11 ), Coulet [ 16 ] (12 )Pastove [ 17 ] - (13 ),, 14,,,

11 (2 ), (15) [ 18, 1 9 ],,,

12 ) 1. (1 ), ( 16 ) ( Reg/ Rel ), Rel [ 20 ],, Rel Reg/ Rel,,,,, 0.2cm ( Pseudomonas ovalis),, [ 21 ],,,, [ 22, 23 ], B rev ibacterium ammoniagenes, NADP [ 24 ] ( 17) 18 [ 19 ],,,,,,, 19, - [ 2 ] (2 ),, (0.30.5cm, 25cm), [ 1 6 ] 2/ 3,1/ 3, 72100r/ min 8

13 , [ 2 0 ] Gluconobacter subox y d ans), ( ),,, Michaelis-Menten, ( PFR ) ( CS T R), [ 21 ] : x S0 - Km ln( 1 - x) = ke t V PFR: x S0 - Km ln(1 - x) = ke 1 q CS T R: x S0 + Km x 1 - x = ke1 q x, S0, Km, k, E, t, V, q, PFR, PFR,, PFR CST R, PFR CS TR, S0 Km,, ; S0 Km,, x = 0.99, S0 = 0.1 Km,, CS TR PFR 21, x x S0/ Km, S0/ Km, x S0/ Km, S0/ Km = 0.1, x , PFR 30%, CST R 50%,,,,, PFR, 9

14 CS T R P FR PFR CST R,,,,,,,,, 1.,1980,44(2 ) : ,1982, ( 2) :14 3 Porter M C. Biotechnol Bioeng Symp Series,1972,3 :115 4 Shrier A I. ibid ,1981,26 :924 6 Chibata I et al. P roc IV IFS Fermentation Technology Today, 1974,383 7 Davis J C. Chem Eng August, 1974, 19 :52 8.,1982,33: ,1975, Scot t C D et al. Biotechnol Bioeng,1976,18 : Poulsen L L et al. Method in Enzymology, 1976, 448 : F ukushima S. Immobilized Enzyme Technology 225, Plenum P ress, Brillouet J M et al. Biotechnol Bioeng, 1976, 18: Suga K et al. J Ferment Technol,1978,56: Vieth W R et al. Biotechnol Bioeng Symp Series, 1972, 3: Brillouet J M et al. Biotechnol Bioeng, 1977, 19: Pastore M et al. Methods in Enzymologp,1974,44: a). 10, 1976,81 b).,1978, ,1978,43 20 F ukushima S et al. J Chem Eng Japan,1979,12 : a). 41,1976,596 b). 11, 1977, Murata K et al. Eur J Appl Microbiol Biotechnol,1978,6 :23 23 Ibid, 1981, 11 :72 24 Murata K et al. Edzyme & Microbiol Technol, ,1979, Wang S S. Biochemical Engineering,1981, ( ) 10

15 ,, 1964, ,, 100 % ( 3 000V/ ml ) 50 % 24/, 45%, 50 ( 60), , 60% 1 1,, 11

16 : St reptomyces S P. : : Snam P ro ge tti CPC Intl. Mil es C linton DE A E Novo Bacillu s co agulan s ICI Art hrobact er SP. Gis t Brocades Ac tinoplan es missourenisis (42% GFCS ) 90%,,,, (90% H FCS ) 90% H FCS 42% H FCS 12, ( 55% H FCS ) 2 2 H FGS : High Fru ctose Corn Syrup, % ( ) 25 % 12

17 ,,, , , ,, , , , 1/ 2,, ( Stevioside) -L- -L-,, 68, (6 ) (7 ) (8 ) -,,,,,, ( ),, ph, ( - ), ( lactose intolerance), ,,, (, 4 5),,, 50 ph23,, ( ) 1977, 1 000, 4 1 3( A), 13

18 , - Phe-105 Met-106 3,,, 3( A),,,, p H 78, -,, 3 (B ),,, 3%, - 55, 50,, , RNA RN A 5-5 -,,, 30,, 1/ 6, 1/ 4 14

19 ,,,,,, 3.,,,, 4., 20,,,, 5.,

20 A T P, ATP ( ),,,, ATP,, ATP ATP [ 15 ] A T P, A T P,, ATP, : 1. ( ), ( ) ADP ADP 2.,, ADP, ADP 3.,, ADP 1 ATP 21 ( p H6.8, 25), 1.4 ( p H4 10, 30) 2 ( ph 7, 30) 12 ( p H6.2, 22 ) 1.5 ( p H9, 100), ( ) 3 ( 25, ) ( ) 7 ( 0, ) ( ) 15 (0, ) ( ) 25 ( 30, ) 16

21 ATP ATP 1 A T P, Mura ta K ATP -6- N ADP ATP ADP ATP, Langer R S [ 1 ] A T P,,, AMP [ 6 ] S, ATP ATP 1 S + 10AM P + 10PPi Mg + + AMP, : AMP + ATP 2ADP ADP + AcP AcP =, A T P + Ac Ac = ( Bacillu s brevis ) S,, 40, ATP, S Whitesides G M [ 7 ], 9 000IU 4 000IU, 1 900IU 2 000IU, AM P, 2molATP( 1kg),, V max, Murate K [ 8 ], ATP ( GS H ), ATP (1 )GSH ADP, A T P, [ 9 ] GS H,,,, GS H 37 SV0.1, 8 GSH,,, 17

22 - Murata K [ 10, 11 ] ATP, ATP, ( T40, Mw ), ATP(2 ),, GSH, 2 A TP, Yamazaki Y [ 1 3 ], ATP -6- A T P,, ATP,,, ATP,,,,,,,, ( Chromatophores ) ADP Pi ( ), ADP Grade VL [ 1 2 ] ( Rho- 18

23 dop seudomonas cap sulata -, ATP, Smeds A L [ 13 ] ( R hodospirillum rubrum)4 000 T 500 ATP, ATP, Pace G W [ 14 ] ADP ATP,, AMP ATPY ang H S [ 15 ], 40%, (1 ) Hill ; (2 ) ATP ; (3 ) NAD- P H, Hill, Ochiai H [ 16 ], ( P VA-117 ), A T P 22 / /, ATP 50% Sawa Y [ 17 ], ( Mastigocladus laminos us ) ADP, A T P,, ATP, Pi ADP, ATP, ATP Arkles B [ 18 ], C8, 27,0 ATP,, Kierstan M [ 19 ] ADP ATP,,,, ADP ATP [ 3 ] ATP, 1 2 CO2, 2 A T P, A T P ATP CDP-,, U DP- UDP-, GS H S- -L- ( S-AM ) [ 11, 20, 21 ], N ADP 1. ; : - ( NM P ) ( NTP ) ; NAD 2. A T P, ; : GSH 3. ; : S- ATP 19

24 CDP- CDP-, 3 A T P CDP- 3 A TP CD P- ATP, CM P CDP-,,,, ATP, Ado Y [ 21 ] ( Saccharomyces cerevisiae) ( ), 35 SV0.25, 8, CMP CDP- 50% 60% (4) ATP, A T P 1 400, AMP A T P, ATP ( 2 ) ATP [ 22 ],,, 34, SV0.1, 10, 42 % 20

25 CDP- 2. A T P A T P Ado Y,, ATP 5 5 A TP 4, 166mmol/ L 43mmol/ L 5 -AMP 4 mmol/ L MgSO4 1 mol/ L ( ph8 ) 35 SV0.2, AMP A T P 70 %,10 50 % ATP CDP-ATP 2 21

26 ATP A TP CDP- A MP 43mmol/ L CMP 20mmol/ L A TP 30.1 CDP % 50 % ADP/ A TP ( mol/ m ol ) 2 3 A T P mmol/ L mm ol/ L ( h) 5 5 A T P ( / h ) A TP = mmol/ L mmol/ L A T P A TP ( mm ol/ L ) ( h), Asada M [ 23 ], AMP,,,, AMP + ATP AMP + ADP AMP + ATP 2ADP + 2ADP + 2Pi 2ATP + 2C2 H 5 OH + 2CO2 (), ATP, ( residence) 18 h, 80 % A T P 3. N ADP ( ) N ADP,,, Muratak K [ 24 ] A T P N ADP,, ATP ( Bacterium ammoniagenes);,, ATP 6 N ADP, 3 N ADP N AD A T P ( p H ),, ATP 22

27 A TP A TP P : S: 3 NADP (m ol/ L h - 1 ml - 1 ) A T P * S. cerevisiae S. cerevisiae B. a mm oniag en es B. ammoniag en es 0 S. cerevisiae B. ammoniag en es : 5 ml 10mL, 30, : 0.5 mol/ L 10 mm ol/ L MgCl mol/ L ( p H7.0 ) 1.0 mmol/ L A MP 10mmol/ L N AD 10 mmol/ L * : 2 ml B. ammonia gen es 3mL S. cerevsiae 10 ml, S. cerevisiae B. ammoniagenes NAD P 23

28 A TP NA DP : 30, SV0.3 : 0.5mol/ L 5 mmol/ L MgCl mol/ L ( p H7. 0 ) 10 mmol/ L N AD1. 0mmol/ L AM P 1.0mmol/ L 15 mm ol/ L 7 8, N ADP,, NADP 4. ATP, ATP N ADP Muratak K, GSH, L- L- L-, N ADP, GS H, (mol/ Lh - 1 ml - 1 ), 0.082, GSH A T P, GS H, A T P,, :, A T P, ATP 1 Langer R S et al. AIChE J.1976,22, ,1981,26,916 3.,1979,43,273 4 Murata K et al. Eur J A ppl Microbiol Biotechnol,1979, 7,45 5 Murata K et al. Biotechnol Bioeng.,1979,21,887 6 Skinner K J. C& EN,1975, 53( 33 ) :22 7 Whitesides G M et al. Methods in Enzymology, ed by Mos bach K Vol , Academic P ress New York,

29 Murata K et al. Eur J A ppl Microbiol Biotechnol,1980, 10,11 9 Yanari S et al. J Biol Chem,1953,201, Murata K et al. J Appl Biochem,1979,1, Murata K et al. Biochimie,1980,62, Garde V L et al. Eur J Appl Microbiol Biotechnol, 1982,13, Smeds A L et al. Enzyme Microb Technol, 1983, 5,33 14 Pace G W et al. Biotechnol Bioeng,1976,18, Yang H S et al. Biotechnol Bioeng, 1976, 18, Ochiai H et al. Agric Biol Chem, 1978,42, Sawa Y et al. A gric Biol Chem,1980,44, A rkles B et al. J Biol Chem,1975,250, Kier stan M et al. Biotechnol Biogen,1977,19, ,1978,56, Ado Y et al. J Solid- Phase Biochem,1979, 4,43 22., 1983, (4) :1 23 Asada M et al. Agric Biol Chem,1978,42, 1533;1979,43, Murata K et al. Enzyme Microb Technol, 1981, 3,

30 ,,, P. Ehrlich, ( ),,,,,,,, ( 5-F U) 5- FU, ,,,,, 5-F U,, ( ) 5 - ( 5 -DFRU) 5-, 5-, 5 -DFRU 5- F U,, 5 -DF UR, 6- ( 6-M P ), 5- FU 5-F U,,,, 1. 5-, 1950,,,, (5-FC), 5- FU, 5- FC,,,, 5- FC, 5-F U, 5- FC,, 26

31 ,,, ,,,,,,,,,,,2 -, B ,, - AMP -AM P AM P, A T P RNA, 10, ( ), mol/ L,,, : , mol/ L, 50% ( EC50 ) 10-4 mol/ L Streptomyces citricolor,,, -6 -, ATP GTP,, 6-MP,, ,,, 3. ( Acyclovir ACV ),, DNA ( HSV) 70,, H SV (1),, 27

32 DN A DU TP D T MP DNA H SV VZ V E BV H SV VZ V CM V EBV H SV VZ V E BV H SV VZ V H SV VZ V E BV VZ V H SV H SV H SV VZ V CM V EBV H SV,,,,,,, H SV, ;, ;, H SV ACV H SV, ACV,, ACV, ACV, ( IDU) ( T F T ) ( Ara-A), H SV B ( HBV), ( / ), ACV ACV HSV,, H SV DN A, DN A, ASV (10 3 ), ACV,, BVDU, H SV, BVD U- ACV 420,,, ACV 6- (Desciclovir )6-Cl-ACV ( TK - TK - 4. ACV ), ACV,,, ACV, D H PG, H SV( TK - 28 )( CMV ), DH PG

33 nor CGM P HSV D H PG 10, D H PG nor-cgm P CMV, ACV,,, ( Cyclaradine) Ara-A,, H SV H SV,, Ara-A ( S )-H P MP A, HSV - 1, ( VZV )0.4g/ ml, CMV0.3g/ ml, H SV - 1 ( TK - ), ( 100g/ ml ) ( R )-, ( S )- 1/ 251/ 100 ( ) ( S )- H PM PA ( S )-D H PA-3-0- ( S )-H PMP A 1UD 2 -, BVDU ( ) CED U, 2 - F IAC FMAU FIA U, H SV, CMV HBV ( EBV), HSV,,, ( PFA ),, HSV CMV, DN A 5. RNA RNA,,, RN A,,, RN A (A ), AICA-,, m RNA, ( Selenazofurin) ( Thiazofurin) 6. ( HIV ) ( AZT ),, AZT,, DN A, HIV AZT 20,,, 2, 3 -( ddc)azt, DN A Sanger DNA ddc,, DNA 29

34 ddc,, AZT HIV,,, dd T, HIV, dda ddc 1/ 10, 2, 3 - T,,,,?,, DN A,,,,,,,,,,,

35 ,, 20,,,,, 1 1 ( ) ( 1 7- ) ( 1 3- ) ( B 4 ) ,,,,,,,, R NA D NA,, 6-, IM P AM P, G GM P, R NA D NA, 5-, du M P d T MP, DN A, 5- F U,,,,,,,,,,,,, 31

36 AM P,,,,,, S- L - (, ) B 12 B 12,, ( ) ( ), DN A,,, DN A, D NA 5-, DN A,, d T MP, DN A,,,, ( ), DNA, ( ddi) ( ddc) cam P 8--cA MP, A T P,, GM P, DN A d T, DN A, DN A d T M P, d T M P, DN A,,,,,,,,, 32

37 A,,, U DP-, CDP- A T P C TP U TP G TP ( NAD ) ( NAD P ) F AD ( ) 5 - ( ),,,,,,,,,,,,, B 2,,,,, A,,,,,, (, ), ( Poly I : C) ( Poly AU ) Isis, 2105, 20, mr NA, m RN A, T, RN A, ( ) 33

38 ,, RN A, A G C U,, ATP cam P A, 2 3 -, 1989 FDA ( A1 A2 ), A1 ; A2,, A T P,, 3050, ATP, 6-6-Cl-, 5 -DFU R F UDR UDP- U T P :5 -DFU R,,,,, UDP-,, C T P CDP ,, CDP- ( ),, 8,,, ( ) (1 7- ) ( 1 3- ) 20 30, 40,, 60, 20,,,, : FDA AZT ddc ddi ; :,,, 20, mrn A,, mrn A,,, 34,

39 ddc 60 DNA,, Sanger DN A,,,,,,,,,,,

40 ,,,, 1992 ( )( DNA) ( RN A) ( ) ( ADP ), 20 DN A DNA, ; DN A C,,,,, DNA RNA,,, DNA 1981 DN A, PCA 1, 3- DNA RNA,,, ph48,,,,,,,,, E, E ;,, A B C,,,,,

41 ,,, ;, 20, 1978, 1992 ( RN A)( DNA), DNA ( dam P ) ( dt MP ) ( dgmp ) ( dcmp ) RN A ( AM P ) ( UMP) ( GM P )( CMP ), ( ), DNA,, , DN A, DN A RNA, DNA, RNA( trn A) RNA ( mrna ) RNA ( rrn A) trna ,,, rrn A, mrna mrn A 10 6,, DNA, rrn A,, (40% )RN A(60% ) rrn A 37

42 ( ATP), (camp ), ( NAD) ( NADP ) A ( CoA),, 1., DNA RNA,,, 1/ 100,,, Rovesti M. Galli, 0.5%,,,,, 2. DNA RNA,,, DNA 1981 DN A DN A,, PCA, DNA RNA,,, ph48,,,, 4. Gilchrest 2880 DOPA 2, nm ; nm,,,,, 5.,, DNA E, E, ADP,, 38

43 DNA,, RNA,, 1 1 RNA ( LD 50 ) 5 900mg/ kg 6 000mg/ kg mg/ kg 2 000mg/ kg 7 500mg/ kg,,, 200mg/ kgday 30,,,,,,, 6 6, RN A 6, RN A %, 0.5%, min,, 12,,,, ( 2025 ), : :,, :,,, :,,,,,,,, 2. DNA 36% DNA,,,,, 39

44 DNA 1%, 5% DN A,,, 10%, 30min, , 0.8gDNA C,,,, 15, 5., RN A,,,,,,, 1. ( DNA RN A) 2. ( K N a ),,,, 3., DN A, AB C E, RN A DN A RN A,

45 - -( Carrageenan) 3, 6-3, 6-,, , 6-,, 3,6-, - - -,, ; -,, ; -,, - -D--4-3,6- - -D- C1, 3 C1, 4, , ,, -,,, ; ; -, +,,,,,,,, -,, g/ cm 2 (1.5 % ), 85% -, ( 30 ), -,,,,,,,, 1990, % ( ), 30%, 10% ( ) 3 000, 1 000,, 25% 30%, -,, FA O/ W H O( ),, - 41

46 , -,, 1.5%,,,,,,, - -, -, 0.05 %,, -,, 0.83% % 0.33 % 60mL, 8520min, 7,,, 0.67 % % 1.33 % 2.,,, -, 70, 25,,,,,,, 3. -,,,,, 0.03 % 0.5% 4.,, 2/ 10 6,,, ;,, ;, % 5%,,,,,,, 42

47 2. -,,, -, ,,, -,,,,, 20% 35 % - -, 0.5% 3.0%,,, 0.1% 3.0%, -,,,, NaCl,,, L- L-, - - ATP, L-, - -, 2 760g/ cm

48 A T P :, A T P, 300L, 2.5t ATP/ a : ATP ( ATP) Adenosine triphosphoric acid, : 1929 Coh mann Fis ke, ATP, ( ADP ) ( AMP ), A T P ATP, ; AMP ; 20 60, AMP ATP ; 20 70,, [ 1 ] 20 80, ATP [ 2, 3 ],,,, 300L, 2.5 ATP 44 1, ADP ATP

49 , : C6 H 12 O5 + 2ADP + 2Pi 2C2 H5 O H + 2CO2 + 2ATP + 2H 2 O Pi AMP ATP, AM P, AM P ADP,,, AM P A T P 14, 1 ATP,,,,,, ATP ATP, ATP A TP 1979 [ 2 ], AMP AMP ATP 70%, 50% Asada [ 3 ], 18h, 80 % AM P ATP [ 4 ],, AMP A T P, [ 5, 6 ] 6%, 2mm3mm,,, AMP ATP 100mL 40mmol/ L 150mmol/ L 8 mmol/ L 250mmol/ L p H ml, 25g, 35 2h, 96 % 10, A T P 90% L, 2/ 3,,, 0.4m 3 / h, , 20mmol/ L, 10, ATP 90% ,,, 45

50 A T P 3 3 A TP 3 ATP : (1 ) 5t ATP 20m 3 300L 25t (2 ), 250t :,, 80 % 90%, ( AMP ), ATP (3 ), ATP, 4,,,,,,,,,,,,, ATP 14 14,,,,, 300L, 2.5t ATP, ATP, 46

51 S,,, 1.,1979, (6) :75 2 Y. Ado etal. J. Solid- Phase Biochem,1979,4( 1) :43 3 M. Asada etal. Agric Biol Chem,1979,43, ,1983, ( 4) :1 5.,1991,17 (4) :421 6., 1993, 25( 1) :

52 ( Ribavirin ) 1- -D -1, 2, ( 1- -D-ribofuranosy1-1, 2, 4-triazole-3-carboxamide), :,, ( )1, 2, 4--3 ( TCA),,,,,, T CA, 28d, : ( 1 ) 2040, ( 2), ( 3 ),,, (4 ), T CA -1-, p H59, 050,,,, -1- ( R-1- P ),, : ( 1), (4060), ;, ( 2 ),, ( 3) ( 4 ), (5 ), 48

53 [ 1 ] 17, ( Geotirchum can didum) O UT 4027( Stemp hy lium botr yosum var. botr ytis) A H U 9130, 26.82%,, % 12, ( Micromonospora pu r purea) IFO 13150, 57.29%,, 27.16% ; ( streptom yces g riseus) IFO 12875,, %,, 44.01%,, 7.23% ; ( S. olivaceus) IFO 12805,, 40.23%,, 40.14%, 24.20% ; ( S. phaeochromogenes)i FO 12898,, 31.24%,, 45.55%,, 23.34% [ 2 ] ( Candid a tropicalis )A TCC ( Saccharom y ces cerevisiae) AT CC 2601, R-1-P,6010h 1 %,,, 34, ( Enterobater aerogenes) AJ [ 3 ] ( B revibacterium acet y licum ) AT CC 39311, ATCC 954, A T-6-7 [ 4, 5, 6 ] AJ 11125, 61 % 62 % ; 81%,, 56% ( Bacillus meg aterium) AJ 3284 [ 4 ], 36% ( Er winia carotovora) AJ 2992 [ 4 ] 91% ( NPase) ( 1),, 1 E. C ( Pyrimidi ne- nu cleosid e phosphorylase ; PyN Pase) E. C ( U ridin e phosphorylase; U Pase) E. C ( T hy midine phos phorylase, TPase) E. C ( Purin e-nu cl eosid e pho sphorylase ; P N Pase) N Pase PyNase PNPase PyN- PaseUPase T Pase;, () 49

54 ) ( ),, 1. PyNPase PN Pase [ 3, 4 ] 2. PN Pase [ 4 ] 3. PyNPase PNPase 1. PN Pase (1 ) ATCC [ 8 ] 2 10L, KO H ph7.2, ATCC 39311, (F ernbach 500mL 30 20h ), 10L 3020h, 12h 20g/ L, 500g, 10mmol/ L,,, TCA 11, mmol/ L, ph K OH 7.2, 70,,, TCA 40mmol/ L,, 40g, 24h 68 % 70 %,, 500mmol/ L, 24h 95g/ L, 65 %, 2 68% 80 % 2 ( g/ L ) ( g/ L ) 20 K 2 H PO 1 14 K H 2 PO MnC l 2 4 H 2 O ZnC l FeCl 3 6 H 2 O 0.01 MgCl 2 6 H 2 O 0.25 CuC l 2 2 H 2 O CaCl 2 2 H 2 O CoCl 2 2 H 2 O Na MoO 1 2 H 2 O P2000 * 2 50 MgSO 4 7 H 2 O 0.75 B B 6 H Cl B d * Dow Chemical, Midla nd Michiga n, U. S. A. (2 ) ATCC 954 [ 6 ] ATCC 954 P NPase, 300 mmol/ L T CA ; 300mmol/ L K3 PO4 ( ph7.0 ), 65, 96 h, 229mmol/ L 100mmol/ L TCA,60, 24 h 74mmol/ L, 74%,,, 68.6 %,, 50

55 , 8.6%, 11 %, 0.90% 2. AT-6-7 [ 7 ] PyNPase PN Pase TCA mmol/ L ( 200mmol/ L), mmol/ L ( 300mmol/ L ), mg/ ml( 230mg/ ml), 6065, ph68, 2436h, ( H + ),, -,,,,, : 1.,, ( 3) 3 ( % ) , ( 4) 4 ( % ) IM P AM P GM P CMP U MP R-1-P , 10 88% (5) 4.,, 62.89%, 51

56 65.44% 5 ( % ) , , H ideyuki Shirae, et al. Agric Biol Chem, 1988; 52( 5) : ,.,1991; (119) : USP H ideyuki Shirae, et al. Agric Biol Chem, 1988; 52( 7) : USP

57 * : : 5-, 6 ( thymidine, 1)2 - (2 -deoxythymidine, d T ), 1-( D- )-5- [ 1- ( 2-deoxy- -D-ribofuranosyl)-5-methyluracil, ( AZT ), AZT, 1 [ 1 ], 1 1 DNA 4, 2 - ( du) Mannich, 4-, Raney 1 [ 2 ] [ 3 ],, DNA DNA,, 1 DN A 1, 2 - ( da) 2 - ( dg) 2 - ( dc ), da 2 - ( di),, [ 4 ], 1 Naylor [ 5 ] ( B revibacterium helvolum) NCIMB , 7d, g/ L 1 25,,, 2 - ( thymine, Thy ), 1 [ 4, 69 ], 5- ( 5-MU) [ ], 5-MU 2 -, 1 [ 15 ] DNA (2 - ) 1 [ 4, 6 ] 53

58 5065) MU : (1 ) : P N Pase( U Pase, PyN Pase) 2 - ( ) + 2- ) + ; (2 ) : -1- ( ( -1- ) + Thy T Pase( P ynpase) 1 (5-MU) + ( 1) ( PNPase, E. C ) ( PyNPase, E. C )( UPase, E. C ) ; ( 2) PyN Pase ( T Pase, E. C ) 1.1 ( G)dG (1 ), PN Pase,,, ( 1), T hy (2 ) 5-MU 1 [ 6, 1 0 ] 1.2 ( A ) (I) da di A da I di, ( 1 ), P NPase, ( 1 ) ( 2 ), ( 1) (2 ) T hy, P NPas - e PyNPase T Pase, 1 [ 79, ] da PNPase, ( 2) [ 4, 6 ] 1.3 ( C) ( U) dc du C dc U du, ( 1 ), UPase(PyNPase), (2 )T Pase, 5-MU 1 [ 4, 5, 7, 10 ] 1.4 DN A ( 2 -) DNA 1, da dg dc, da didg da di dc , 1 [ 4, 6 ] ( E rwinia carotovora) AJ-2992 G T hy 5-MU [ 10 ] 4 10min, 50mmol/ L Tris-H Cl ( ph8.5 ), 50mmol/ L T ris- HCl ( p H8.5 ) 20mmol/ L K2 H PO4 100mmol/ L G 100mmol/ L Thy ml 50mg, h, 3 24h, 5-MU mmol/ L 2.2 ( Bacillus stearothermop hi lus) J TS 859 I di T hy 5-MU 1 [ 7 ] 54

59 2.2.1 ( g/ L)I 5.36 T hy 2.52 K H2 PO4 g,10min), 4.08 K2 H PO ( ph7.0 )10mL 50mL, 130mg, 65 3h, 5-MU1.57g/ L( 6.1mmol/ L) 0.25, 6.6mmol/ L 92 % mg, 20mmol/ L ( ph7.0) 0.2mL, ( mmol/ L)dI 25 Thy ( p H7.0) 0.8mL, 60 2h, 4.62 mmol/ L mmol/ L68 % 2.3 ( B acil lus stearothermop hilus ) J T S859 A da Thy 5-MU 1 [ 8, 9 ] 2.3.1, ( mmol/ L) A5 Thy mg 2U 0.26U, 1 ml ( p H7.0 ), h3.7mmol/ L 5-MU, 74% 58 % da A,, 2.9mmol/ L 1, 2.4 ( Bacillus stearothermophilus) J TS859 P NPase PyN Pase I T hy 5-MU [ 1 2 ], 10mL DEAE-Toyopearl 650 M 10mL, 20mmol/ L ( p H7.0) min,, 20mmol/ L ( ph7.0 ) U 1 700mL 135mm140mm, ( mmol/ L)I75Thy75 20, 60 1L/ h, 5-MU 24 mmol/ L, 17d, 2.5 ( E. col i) BMT-1D/ 1A da dg dc DN A Thy 1 [ 6 ] T Pase P NPase ( 1mL ) p H7.0 90mmol/ L Thy 30mmol/ L 2-0.1% ( ) E. coli BMT-1D/ 1A, E. coli BMT-1D/ 1A Thy da dg dc ( h) ( mmol/ L ) ( ) ( % ) dc dg da DN A ( S picaria v iolaces) BM-105D, ( 1mL ) 10 mmol/ L ( ph7.0 ) Thy 2-0.1% ( ) E, coli BMT-1D/ 1A,

60 E. coli BMT-1D/ 1A Thy DNA 1 T hy ( 1 ) ( mmol/ L ) ( mmol/ L ) ( h) 1 ( mmol/ L ) ( h) 31( 9.1) ( 18.2 ) ( 36.4 ) da dg dc di DNA Thy 1 [ 4 ] g/ dl ( mmol/ L)dA(dG di )20 Thy ( ph7.0 )10mL, 6024h 31 da(dg di) Thy 1, da dg di Thy I 1 ( mg/ d L ) da dg di A T CC - 954( Bre vib act erium acetylic um ) A TC C ( Cellulomon as fla vige na ) A T CC - 373( Coryn eb act erium x erosis ) A T CC ( Mycobac teriu m lac ticolu m ) FE RM - P7048 ( Sarcin a alb a) I FO ( St aphylococcu s citreus ) mmol/ L dc da, 34 dc Thy 1, dc Thy 1 1( mg/ dl ) FE RM - P9468 (Bacillus sph aericu s ) 248 A T CC - 373( Coryn eb act erium x erosis ) 87 EFRM - P9736 ( Esch erichia c oli) 152 A T CC ( E nterobact er clo acae) 130 I FO ( St aphylococcu s citreus ) mmol/ L ( ph7.0 ) 50mmol/ L Thy DN A [ ( mg/ dl) : 1 282, da 97, di 124, dc 121, dg 157 ] 10mL, IFO FERM - P g/ dl 608h mg/ dl, 1, I A G C U, T hy 5-MU, 1,, 5-MU 1 56

61 Thy DNA,, da dg dc du 1, 1 DNA ( ), 1,,. ( ),1991 ; ( 119 ) :85 2 Reese CB. J Chem Soc, Chem Commum,1983 : (16) :877 3 Vorbruggen H, K rolikiewicz K, Bennua B. Chem Ser,1981;114 : ( CA 1989;111 :152191) 5 Eur Pat A ppl 1989: ( CA 1989; ) 6 Zinchenko AI, Barai VN, Bokut SBB et al. Biotechnol Lett, 1990; 12 : (CA 1989 ;112; ) 8 H ori N, Uehara K, Mikami Y. Biosci Biotech Biochem,1992 ;56 : ( CA 1993 ;118:232397t) 10 Ishii M, Shirae H, Yokozaki K. Agric Biol Chem, 1989; 53 : Hori N, Watanabe M, Yamazaki Y et al. Agric Biol Chem,1980;53: Hori N, Watanabe M, Sunagawa K et al. J Biotechnol, 1991; 17 : Hori N, Uehara K, Midami Y. Agric Biol Chem, 1991;55: ,,.,1993 ;23 :1 15 Robins MJ, Wilson S. J Am Chem Soc,1981 ;103:

62 (DN A RN A),,,,,,,,, ; ; ( de nove, ),, Van Bur Kulkarni,,,, Sonoda, 14 C RNA, 1 70%, 15 % 4,, 47%, 40 %, 8% ( Salvage), 1 4 C DN A, 4, 10%,, DN A 14 C RNA, 4, 30 %,, AM P,, 70%, ( 3 7.1% ) 8.1% 24 57%,, ( DNA RNA),,,, 58

63 de Novo ),, (,, ),,, ( ),, [ 8 ],, ATP,,, DNA RN A,,,,,, DN A,,,,,, ( ),,,, ( ),,,,,, ( T P N),, : OG-V1( = ) ( ), T PN OG-V1, DN A RNA OG-V1 T PN,, RNA DN A, ( ) RN A, W-3 (D H A, E PA ), W-6 ( - ) H,,,, 59

64 OG-VI,,,, RN A, GI,,, m RNA, mrn A, OG-V1,,,,,, DNA,,, 6,,, (),,, de novo, Salvage,, de novo, de novo Salvage,, M TX, MTX,,,, M TX galvage, MTX, galvage,, 5- F Ura F Ura, F Ura,, Salvage, denovo,,, de novo,,, de nove Salvage,

65 ,, ( ),,, DNA RN A,,,, 5 -,,, : ( Acyclovir ),, : FDA ( HIV) ( Zidovudine, AZT ) ( didanosine, ddi ) ( Zalcitabine, ddc) ( Stavudine, d4 T ) ( Lamivudine, 3 TC) ( V alciclovir) ; ; ( Vidarabine) ; ( Ribavirin ),,,,,,,, -, DNA RN A,,,,,,,,,, ( ),,,,, 61

66 ICN, 85%, DN A, 1977, FDA,, 400,,,,,, FDA 5 ( H IV ) AZT ddi ddc d4t 3 TC 5 -, DNA 3 -, DN A DN A RNA HIV AZT d4 T DNA,,,, DN A, da dg dc dt, RNA, 5-, dt, d4 T 2. ddl Eastwood ddu, dda, ddi,,,,,

67 :, N-, : N- HIV (zidovudine, AZT )( didanosine, ddi) (zalcitabine, ddc) ( stavudine, d4 T ) (lamivudine, 3 TC), ( vidarabine, Ara-A) ;, 50%,,,,, -, : ( GR) ( IR), GR,, 3 45% 48% GR, 85%, 66% [ 1 ] ( 1 ), N- ( 2 ), 1 1 1, 1 ( ), ( )-1-, : Rib (d Rib) - [1 ] + Pi Rib (d Rib) -1-+ [ 1] Rib (d Rib) -1-+ [ 2] Rib(dRib) - [ 2] + Pi Rib: D-, drib: 2 - -D-, : 1 ( P NPase, EC ) ( PyN Pase, EC )GR ( IR ) ( GPase, EC ) ( A Pase, ) ; ( UR ) ( U Pase, EC ) ( dt ) ( dt Pase, EC ) [ 2 ] PyN- Pase, UR, PyN Pase 63

68 , 1.1 1, 1 : P NPase ( AR ), IR GR, 3,, ( % ) A J1442( A T CC954) T Q952 GRTCA A J2992 T CA 91 4 GRT A J11125 Ara- UA Ara- A 76 6 du ( ) T d T 55 7 A T CC373 di( da dg) T d T 54 8 I FO12083 d U, TF M U T FT 48 9 T CA:, T :, A ra- U:, A :, du :, TF M U :, TF T : ( triflu ridin e) 1.2 P NPase, : IR T CA, U tagawa [ 10 ] AJ11125 PN Pase, TCA, 4020 %,, IR Shirae [ 4 ],, 91% 1.3 1,,,, Ara-A, 60 [ 6 ] 60 70, % [ 11 ] , 1,, D-, 1-1-,

69 , (, chemico-enzymatic method s), AZT AZT 5- dt, 1 [ 5, 7 ], AZT, AZT (1) 1 A ZT Ara-A [ 6 ], UR Ara-U, Ara-A (2) 2 Ara- A 65

70 3) ddi [ 12 ] Eastwood ddu, ddi ( 3 ddi 1.5,,,, : Ara-U A Ara-A, A Ara-A, 40%,, 2 2 (EC ) 1, 2: ( ) : drib- [ 1] + [2 ] d Rib- [ 2] + [1 ] () : a) drib- [1 ] + [ 2] drib- [2 ] + [ 1] b) drib- + drib- + c) d Rib- + drib , Smar [ 3 ],, 2 dt, (Dribal), ,,

71 2 ddc N N 6 - HI V HI V TF MU 2- [ 14 C] - T FT 17 dd T ddc HI V , [ 19 ] N 6 - [ 20 ] 2, 2, 3 - [ 17 ], [ 18 ] , dt (dd T) dc(ddc )2 dt (dd T) dc(ddc), [ 2 3 ] 2, 2, dt (ddt )dc 3 3.1,, Vozaki [ 2 ] Amano PS ( Pseudomonas sp ) 5- FUd R 5- T F MUdR 3 -O- Ozaki [ 22 ] , R = CH 3 ( CH 2 ) n, 2-67

72 ( ADA ase) ADAase, ADAase dda ddi [ 23 ], ddi,,,,,, 1,,., 1997;22( 23) :1 2,,., :,1982 : Shirae H, Yokozeki K, Uchiyama M et al. A gric Biol Chem,1988 ;52 : Shirae h, Yokozeki K, Kubota K. Agric Biol Chem, 1988 ;52 : Ishii N, Shirae H, Yokozeki K et al. Agric Biol Chem,1989 ;52 : ,.,1985 ;43:2 7,,.,1996 ;22(6 ) : : (CA 1989:111 :152191m) 9 Ishii N, Kobayashi K, Shiragami H et al. Appl Environ Microbiol, 1989;55: U tagawa T, Morisawa H, Yamanaka S et al. Agric Biol Chem,1986;50: H uavd CN, H utchinsom DW. Biocatalysis, 1992;6 (1 ) : :49493 (CA 1993;118 :232397t) 13 Smar M, Short SA, Wolfenden R. Biochemistry,1991 ;30 (12 ) : PCT Int A ppl 1990:06312( CA 1991 :114:99958a) 15 Haertle T, Carrera CJ, Wasson DB et al. J Biol Chem, 1988 ;263: Heidelberger C, Par sons DG, Remy DC. J Med Chem,1964 ;7: 1 17 Carson DA, Wasson DB. Biochem Biophys Res Commun,1988; 155 : Van Draanen NA, F reeman GA, Short SA et al. J Med Chem,1996 ;39 (2) : Betbeder D, H utchinson DW. Nucleosides N ucleotides. 1990;9 (4 ) : Betbeder D, H utchinson DW, Richards A et al. A ntiviral Chem Chemother,1990;1 (1 ) : Nozaki K, Uemura A, Yamashita J et al. Tetrahedron Let t,1990 :31 (12) : Ozaki S, Yamashita K, Konishi T et al. Nucleosides Nucleotides, 1995; 14( 35) : US 199 : (CA 1992 ;116: 19722x)

73 D- : D-, B2,,, D-, D-, D- : D- D-, 1909 ( Levene)( Jacobs) [ 1 ] D- RN A 20 50,, B2 70 ( Sasajima) [ 2 ] ( TKT, EC ) D- D-,, 80,, B2, D- 1 D- D- TKT ( PPP ) PP P 1 D-TKT D ( RPE, EC ), P PP TKT, D- -5-, D-, [ 3 ] , ( Simonart) ( Godin) [ 4 ] D- 10, D- D- -5- [ 5 ] 1966, ( Saito)( Sugiyama) [ 6 ], 50g/ L 0.97g/ L D-,,, D-,,,, D- -5- [ 7 ] 20 70, ( Sasajima) [ 8, 9, 1 0 ] 69

74 ,, D-, L-,, shi5, shi7, Gluc34, D-, 6 100g/ L 35 g/ L D-, TKT RPE TKT D-, RPE D-, D- D-, TKT D-, ( Sasajima ) [ 11 ], TKT, D- ( GDH ),, D-65g/ L, ; 72.3g/ L GDH, GDH, D- D- : ( D- ) ( PPP ) 2 70

75 D- ; ; ; ; 6- ; 5- ; 5- ; D-, D- D-D- D-, D- 1992, Miyagawa [ 1 3 ], D-D-,, D-D-I FO15138, 160g/ L D- 39g/ L 62g/ L [ 14 ] AT CC 31093,, ( 90g/ L ), RE5 RX13, g/ L118.8g/ L, GDH, ( GAK ) D- [ 3 ] D- ( g/ L ) D- ( g/ L ) ( h ) A T CC Yone da & Sasa jima( 1969 ) A T CC Yone da & Sasa jima( 1969 ) A T CC Sasa jima & Yon eda( 1971 ) A T CC Sasa jima & Yon eda( 1974 ) A T CC Sasa jima & Yon eda( 1974 ) A T CC Sasa jima e t al.( 1975 ) A T CC Sasa jima e t al.( 1975 ) B. s p E MP A sai et a l.( 1978 ) A T CC Kint a ka et al.( 1986) A T CC Kishimoto e t al.( 1990 ) IF O Miya gawa et a l.( 1992 ) A T CC / 100 * Dewulf e t al.( 1996 ) ATCC (RX13) (1996 ) [ 14 ] A T CC Dewulf e t al.( 1997 ) 9704 D / 50 * * (1999 ) [ 16 ] TJ ( 1999 ) [ 17] * 100g/ L D- 100g/ L D- * * b100g/ L 50g/ L D- 71

76 D-, 3 TKT D-TKT,, D-, 3.1 TKT D-,, D- D- [ 10 ], ( PE P ), P EP D- glc ( -, EC ) [ 15 ] 3.2 TKT, D-( D-, D ) D-, D-, D-, L-, D- (D-, D- ) (,, ) D- TKT, D- -5 [ 18 ] TKT ATCC D- D-, D-, L-, D- D-, TKT ATCC 6051,, ATCC D- PP P D-, D- -1, 6- [ 19 ] 3.3 TKT,,, SP01 SP10 TKT (), [ 2 0 ] TKT SDS, [ 15 ] 4 D , L-, L-, L- (0.1g/ L), 1.7% 10 % 37, p H 72

77 8.0, [ 15 ] 2 ( g/ L ) D , D- D- D- D- D-, D-,, [ 15 ] ( Sasajma) [ 15 ],,,, Kishimoto [ 21 ],,, D- 20 %, ( ) 3, De Wulf [ 22 ], D- (60g/ L) D-, ( Sasajima) Yoneda [ 2 3 ], (, ), D- 40 % De Wulf [ 2 4 ] 2g/ L, ATCC ( 2, 3- )90%, D-40 %,, ph p H,, A P T A P T D- PPP, D- -5-, D- 3 3 ( g/ L ) D- 150, 200 ( N H 4 ) 2 SO MnS O Ca CO TKT, D-, D-,, / ml D- [ 9 ] 73

78 D P PP,,, ( D-), ( 2, 3- ), D- [ 22 ] AT CC D-, ph 6.0p H,, p H, D-, D-, [ 22 ], ph, D-, D-,, ( ) [ 3 ] D-, De Wulf [ 19 ] (D-, D-, L- ) D-,, D-50%, D-Asai [ 25 ], De Wulf [ 3 ], AT CC ,,, [ 26 ], -,, 40.5 g/ L 5g/ L, D- 65g/ L 90.6 g/ L Asai [ 2 5 ] D-, 36.6 D-,, D-,, 3 3 D- [22 ] 37, :72h, : A T CC D- RNA D-, D- [ 27 ] [ 28, 29 ],, D- [ 30, 31 ] 74

79 Saito ) ( Sugiyama) [ 6 ], ( ), D-, D- ( Sasajima)Yoneda [ 9 ] TKT,,, ( Amberlite I R 120, Amberlite IRA 400 ), D- Hough [ 32, 33 ] D- [ 34 ],, 732, D-315, HD-1,, 20% 30%, 6,, D-, D-,, [ 35, 36 ] D-, [ 37 ], [ 3 7 ] [ 38 ] D-,,,,,,,,, D- 1 Levene P. A. & Jacobs W. A. Uber. Inosinsaure. Ber.,1909,42: Sasajima, K. D-Ribose. In : Microbial Production of Nucleic Acid-Related Substa nces, ed. K. Ogata, S. Kincshita, T. Tsunoda & K. Aida. Kodansha Ltd, Tokyo and John Wiley & Sons, New York. London, Sydney, Toronto.,1976,pp De Wulf P. & Vandamme E. J. Production of D-ribose by fermentation. Appl. MIcro. Biotechnol., 1997,48 : Simonart P. & Godin P. P roduction de pentoses, d acide 2-ceto gluconique et d acide glucuronique par Penicillium brevi-compactum. Ghim.,1995,60 : Suzuki T. et al. Bacterial accumulation of ribose and ribose-5-phosphate. J. Gen. Appl. Microbiol., 1963, Saito N. & Sugiyama S. D-Ribose forma tion by Pseudomonas reptilivora. Agric. Biol. chem., 1996, 30 : Kinoshita S. et al. P roces s of producing D-ribose-5-phosphate and D- ribose through fer menta tion., 1967, US patent Sasajima K. et al. Carbohydrate metabolism mutants of a Bacillus species. Part. Isolation of mutants and their inosine formation. Agric. Biol. Chem.,1970,34: Sasajima K. & Yoneda M. Carbohydra te metabolism mutants of a Bacillus spectes. Part. D-Ribose accumulation by pentose phosphuate pathway mutants. Agric. Biol. Chem., 1971, 35 : Sasajima K. & Yoneda M. D-Sedohe pt ulose 7-phosph ate: D-glyce raldehyde 3-phospha te glycolalde-hydet ransfer ase and D- ribulose 5-phosph ate 3-epimerase m utan ts of a Bacillus s pecies. Agric. Biol. Chem.,1974,38 : Sasajima K. et al. Method for the production of D-ribose. 1976, US patent

80 Yokota A. et al. Reactivation of inactivated D-glucose dehydrohenase of a Bacillus species by pyridine and adenine nucleotides. Agric. Biol. Chem.,1979,43 : Miyagawa K. et al. Method of producing D-ribose. 1992, European pa tent A1 14,, Sasajima K. Yoneda M Production of D-ribose by microorganisms. In : Vandamme EJ (ed) Biotechnology of Vi5tamins, Pigments, and Growth Factor s. Elsevier, New York. 1989,pp ,2000,26(1 ) : ,1999,28( 1) :12 18 Sasajima K. & Kumada T. Change in the regulation of enzyme synthesis under catabolite repression repression in Bacillus subtilis pleiotropic mutant lacking transketolase. Agric. Biol. Chem.,1981,45 : De Wulf P. et al. D-glucose does not catabolite repress a transketolase-deficient D-ribose producing Bacillus subtilis mutant strain. J. Ind. Microbiol.,1996,17 : Sasajima K & Ku mad a T. Deficie ncy of f lagellation in a Bacillus subtilis pleio tropic mu tan t lacking trans ketolase. Agric. Biol. Chem.,1983, 47 : Kishimoto K. et al. Production of D-fibose. 1990, US Pa tent De Wulf P. et al. Optimization of D- ribose production wit h a t ransk etolase-affected Bacillus sub tilis mutant strain in glucose and gluconic acid-based media. J. Appl. Microbiol.,1997,83: Sasajima K. et al. Method of producing D-ribose.1975, US patent De Wulf P. & Vandamme E. J. Specific organic acids enhance the D-ribose productivity of a transketolase-defective Bacillus subtilis strain. J. Chem. T echnol. Biotechnol., 1997, 70 : Asai T. et al. Kinetic Study on the production of D-ribose by Bacillus sp. J. Ferment. Technol., 1978,56 : ,,,, Ekenstein W. A. et al. Over D-ribose. Chem. Weekblad.,1913, 10 : Berger L. & Lee J. Arylamine-N-glycisides. Part. Arylamine- N-pentosides and complex salt forma tion studies. J. Org. Chem.,1944,11 : Berger L. et al. Arylamine- N-glycosides. Part. Hydrolysis of arylamine- N-pentosides and the preparation of crystalline D-ribose. J. Org. Chem.,1944,11 : Jeanlotz R. W. et al. The chemistry of D- ribose. Adv. Carbohydr. Chem. Biochem., 1951, 6: Overend W. G. & Stacey M. Chemistry of ribose and deoxyribose. In : E. Chargaff & J. N. Davidson (ed.) The Nucleic Acids, Vol., Academic P ress, New York, 1955, pp H ough L. et al. Application of paper partition chromatography to the separation of the s ugars and their mehylated derivatives on a column of powdered cellulose. Nature, Lond., 1984,162: Hough L. et al. Quantitative analysis of mixtures of sugars by the method of partition chromatography. Part. The separa tion of the sugar s and their methylated derivatives on columns of cellulose. J. Chem. Soc.,1949: , Zimmer H. G. The oxidative pentose phos pha te pathway in the heart-regulation, physiological significance and clinical implica tions. Basic Res. Cardiol.,1992,87 : Zimmer H. G. et al. Ribose intervention in the cardiac pentose phosphate pathway is not species specific. Science., 194,223:

81 Gros s M. et al. Ribose administ ration during exercise : effects on subst rates and products of energy metabolism in healthy subjects and a patient with myo-adenylate deaminase deficieny. Klin. Wochenschr.,1991,69: Chan M. F., Hsiao C. N. A versatile and stereospecific synthesis of a dihydroxyethylene dipeptide isostere of renin inhibitors from D-ribose. Tetrahedron. Let t., 1992,33 :

82 :, ATP, ATP, ATP, : : Q528.2 D- ( D-Ribose), 1909 ( Levene) ( Jacobs ) 20 50,80,, [ 1, 2 ] D-B2,,,, ;,, [ 3 ] 1-6- ( G-6- P ) G-6-P, 6- ( G-6- PD H G- 6- PD) : ( 1) N ADP H ( GSSG) (GSH ) ( 2) -5- ( R-5-P ) : R-5- P R-5-P 5- ( PRPP ), U T P ( ) 2, G-6-PD, PRPP,,, 78

83 ( ), [ 4 ],, G-6- PD 6-PGD(6-, ) ; G-6- PD G-6- PD,, PRPP,, ATP [ 5 ] :, ATP, ATP,,, ATP PRP P, ATP, ATP, PRPP AMP IMP PRPP : ( 1 ) G-6- PD, ; ( 2 ), PRPP,, 1 [ 4] ( U/ mg ) G-6- PD H 6-P- P < < < < < < < , R-5- P PRPP,, ATP, ( 0.5mmol/ L ), min, ( AN, ATP + ADP + AM P ) 10 ; ( 25mg/ kg) ATP, ( 100mg/ kg ), A N, PRP P, ( 200mg/ kg ) A T P ;, 15min, 250mg/ kg 20min, 79

84 A T P ; 2h, ( 0.1 mmol/ L) ( 15mmol/ L), ; 15 min, 200mg/ ( kgh) 72h, 12 h ATP ; 15min, 200mg/ ( kgh) 24h,, ATP A N ;, 200mg/ ( kgh ) ATP, ATP, 15min ATP 40 %,, ATP, ADP, 3d,, A T P 12h, 60h [ 3 ], ATP,, 24h, 0.9 % NaCl ATP 24h,, A T P [ 6 ],,,, - -, ATP 24h, 3d, ATP, [ 7 ] ( MAD) [ 8 ] MAD,, 29%,, g, McArdle s (, V ) 60 W 100W, [ 9 ], h, 2 [ 10 ] 5,, ( Verapamil), - ( Motoprolol), [ 1 1, 1 2 ] 80

85 ATP, 5h,, ATP, ( ) ( ) 5h ATP PRPP (12h ), ATP [ 13 ],,, ATP, 1 min [ 14 ] 5d, ATP, [ 1 5 ] D-,, 1 De Wulf P, Vandamme EJ. P roduction of D-ribose by fermentation. Appl Micro Biotechnol, 1997,48: ,. D-.,2000,30( 1) : Zimmer H-G. The oxidative pentose phosphate pathway in the heart : regulation, physiological significance, and clinical implications. Basic Res Cardiol, 1992, 87 : Zimmer H-G, Ibel H, Suchner U, et al. Ribose intervention in the cardiac pentose phosphate pathway is not s pecies specific. Science, 1984,233: Reimer KA, Hill ML, Jennings RB. Prolonged depletion of AT P and of the adenine nucleotide pool due t o delayed resynt hesis of adenine nucleotides following r eve rsible myoca rdial ischemic injury in dogs. J Mol Cell Cardiol, 1981,13: Zimmer H-G. Normalization of depressed heart function in rats by ribose. Science, 1983, 220 : Zimmer H -G, Martius PA, Maschner G. Myocardial infarction in rats : effects of metabolic and pharmacologic interventions. Basic Res Cardiol, 1989,84: Patten B M. Beneficial effect of D-ribose in patien t wit h myoadenyla re d eaminase deficiency. La ncet, 1982,1: Wagner D R, Zollner N. McA rdle s disease: Successful symptomatic therapy by high does oral administration of ribose. Klin Wschr, 1991,62:92 10 Perlmut ter N S, wilson R A, Angello D A, et al. Ribose facilitates thallium-201 redistribution in patients with coronary artery disease. J N ucl Med, 1991,32: Mauser M, Hoffmeister H M, Nienaber C, et al. Influence of ribose, adenosine, andaicaron the rate of myocardial adenosine tripnosphate synthesis during reperfusion after coronary artery occlusion in the dog. Circ Res, 1985,56 : Zimmer H -G, Zierhut W, Marschner G. Combination of ribose with calcium antagonist and -blocker treatment in closed-chest rats. J Mol Cell Cardiol, 1987,19 : Zimmer H -G, Schneider A. Nucleotide precursors modify the effects of isoproterenol. Studies on heart function and cardiac adenine nucleotide content in intact rats. Cire Res, 1991,69 : Lee RGL, Springer C, Kasulis P, et al. Nuclear magnetic resonance assessment of adenosine triphosphate ( A TP ) dynamics in ischemic mouse livers perfused with adenine and ribose. Invest Radiol, 1987, 81

86 McAnulty J F, Southard J H, Belzer F O. Improved maintenance of adenosine triphosphate in five-day perfused kidneys with adenine and ribose. Transplant P roc, 1987,19:

87 C T P : A T P, -,, CT P,,,, GTP U T P damp dgm P dc T P d T T P : CT P ATP - 5 -Cytidine triphosphate( CT P ), : CT P,,, 1 C T P ATP() ATP,, ; ( ) ( Synthetase), CC CO CN CS, ATP, : A + B + ATP + H2 O A - B + ADP + P( ADP ) ATP,, A T P:, 83

88 ADP ATP, ( 1) ATP,, 1 A T P CT P -,, ATP, ATP 3 000,,, ATP ATP, ( ATP C T P GTP U T P datp dct P dgtp dt T P ), S- UDP- C T P, A T P ATP, ; Asada ; ATP,, CT P,, 88%, 7 h 2 -,, CT P, 20 80, 84

89 , 1 008kg/ cm 2 = kg/ cm 2, 3, CM P ( ), CT P 40mmol/ L CM P, 2h, C T P,, , CN A C T P : CMP CT P C T P 3 :,, ;,, CT P, CT P CDP ( ) CM P, ;,, 3 C T P, CM P C T P, C T P,,, GTPU T P damp dgm P dc T P dt T P

90 A T P,,, ( ) ( synthetase), CC CO CN CS, ATP : A + B + ATP + H2 O, : A - B + ADP + Pi ATP,, A T P :, ADP, ATP, ATP,,, ATP 5 -( 5 -AM P ) A T P, ATP, ATP : 86 1 A TP

91 , ; ATP, ATP, ATP ADP,, 80% --, 10, ATP, ATP 90%,, ATP 1985, ( C. am monia genes), ATP,, 2, 3 2 A TP 3 A TP 4,,,, ATP 87

92 , ATP, ATP 22 43mg/ ml (1 ) GM P, GM P, ATP, GMP (2 ) IMP, A T P,,, 5 IM P 88

93 IMP, ATP,,, 60% 80 %, C T P,, UTP ATP,, 7 A T P CDP

94

95 ( CDP- )Kennedy [ 1 ],,,,, ( 1 )1970 CDP- [ 2 ], 5 -CM P CDP-,,,, CDP-, 1 CDP-,, - [ 3, 4, 5 ], CDP-ATP, : 5 -CMP ( 89% ) ( 70% ) : (18B ) 93

96 [ 6, 7 ], 1mm, , 720 ( EDTA- 720, ph7 ), 24, H2 O2 (2.4 30% H 2 O2, 720 ), H 2 O2, ( , p H2.5 ) 80r/ min 3, ( r/ min ),,,, 12% NaO H ph 7, 2., 40 ( 4 ), 90, ph2.53.0,, 34, 60 ( 12 ),,, 0.1mm, ph mol/ L, ph8.5 1% 6, 1, 3. CDP-,, 2.550, (2), 6B, 34 24,,, SV = 0.1, 34,, 24, : 400mol/ L, 5 -CMP 20mol/ L, 100mol/ L, MgSO4 7H2 O 50mol/ L, ph mol/ L 4. 1% 2, (2mm ), 10 : 0.05mol/ L ph9.3 30, 8, Whatman 4, 10, : ph9.3, mol/ L, 10, 90, 2 80,, 5 0.1mol/ L H Cl 1, OD : = CDP-OD2 80 (5 -CM P + + CDP- ) OD % = 100% 94

97 : 72, 7237% = 26.6, 180 (), 18 = 18/ % = 68% :, A B C ( mm ) ( mm ) ( mm 2 ) ( g) ( g/ mm 2 ) A :, B: C :, : (/ 2 ) = () ( 2 ) 2. : 10% 5 % 2.5% 1% 0.5 % , , :, (50 / ) 200, 34, 14,,, 280 CDP-,,, 1%, 6,, : ; 13, 14,15,,, 14,,, 280 CDP-,, : 13, 14, 15 15,, : = 13 ( ) 3. CDP- SV 0.1, 34, 2 10, 5 -CM P CDP-44 % (3 ) 95

98 CDP- 4.,,,,,,,,,, CDP-,,, CDP- 44%, 41.8%, 18 %, CDP-, 1 Kennedy E P et al : J Biol Chem, 1956,222 :193 2 Tochikura T et al: J Ferment T echnol, 1970, 48 :763, Murata K et al : Eur J A ppl Microbiol Biotechnol, 1981,11 :72 4 :,1979, 21:426 5 Ado Y et al : J Solid-phase Biochem, 1979,4 :43 6 U S 2,920,000, Whitesides G M et al : Methods in Enzymology, 1976,44 :246 8 :26,915,

99 ( CDP- ), CDP- [ 1 4 ],, [ 5 ],,,,, 40 / 5,, 1. ph9.2, 0.05mol/ L 2. () 3. : 5 -CM P ( ) ( Fluka AG, Buchs SG, Switzerland ) CDP- ( ) 1. DY-1 ( ) 2. SU V-1 ( ) G () mm75 mm 100mm75mm , p H 9.0,, 769, , 40 60,, 0.6%, 97

100 ,, 2,, p H9.2, 0.05mol/ L,,, 300, 5, 5. SU V-1,,, 5 ph1.0, 2,, 751G 280nm OD 1,, 1 2 2,, 3 CDP- 1 : 1. ( CD P- ) ; 2. ( CMP CD P- ), ; 4. D H CDP-

101 (g) C O.D 280nm * O.D 280 = C * O.D 280 n m 2 O.D CDP- CD P- (g) CDP- (g ) CDP- (g ) ( % ) ,,,,, SU V-1,,,, (1 ) [ 1 ], CDP-O.D280 nm O.D2 80 nm, CDP- 1 :1977, ( 4) : 15 2 : 1976, 3(3 ) : :1982, ( 6) ; 63 4 Tochikura T et al: J Ferm Technol 1970, 48 : Ado Y et al : J Solid-P hasa Biochem 1979, (4) ;

102 ( CDP- ), CDP-, CDP-,, 70% 80 %,, 2530, 30 %, [ 13 ], Dowex 12, DH846,,,, CDP , D H / 260 = 2.1,,,, 45% 1. CDP- [1 ] 2. CDP- ph 3.5,, p H 7.0, ph , 769, 1/ mol/ L0. 006m ol/ L0. 01m ol/ L HC l ph , 769, p H 3.0, p H 3.5,, ph 9.0, p H , 769, 1/ 2 D H mol/ L 0.02 mol/ L H Cl, 20 % 95 % 20 % 95 % 100

103 CDP ( ), ph 9.0 (, ph ), ph , 769 ph , Cl -, (95% H2 O = ), D H846 [ O H - ], CDP mol/ L H Cl, 0.02mol/ L HCl(NaCl), D H846 2mol/ L H Cl 2 mol/ L N aoh CDP- : ( ) ( ) * 82 % 90 % % 90 % D H % 95 % 85 % 35 % * * - 45 % 52 % * * * [1 ] 1 D H846 CDP- 2 DH 846 ( O H - ) CDP- : 1.8cm : 32cm : SV 0.2 : 500mL, D H846 CDP- CDP-, 101

104 ), 1 : 1976, 8(3 )199 2 :1979, ( 5), 40 3 : J Ferm Technol 1970, 48,

105 ( ) :,, p H 58%, KM = mol/ L, : ( CDP- ), 1963, CDP- ( EC ), CT P ( ) CDP- C T P + CDP- + Borkenhagen, L.F.Kennedy, E.P g [ 1, 2 ] [ 3 ], CDP-, CT P K CT M P, (), ; CT P, ;, ;, () 1. p H mol/ L -, DLL-3 (2 800r/ min,0,15min) ;,

106 CDP- [ 4 ] : 1mL 5min,, :0.6 % ( W/ W ),,, 20L 5min ( - ph9.65, 0.05mol/ L, 40 ), SU V-1,, 5mLpH1.0,, 280 nm OD CDP- 3. [ 5 ] CDP-% = CDP- CT P 100 % 4. CDP- OD28 0n m = 100 % OD2 80 nm ml : CT PMgCl2 10mol, 28mol, p H6.5 Tris 50mol, 1g, : 50mL 5 ml,,326h ( ) 1. ph CDP- p H 5.5, 6.5, 7.5, 8.0, ph 6.5, CDP- 1 ph CD P- 2 CD P- 2. CDP- 28mol/ ml, CDP-,, CDP- (2 ) 104

107 CT P CDP- CT P 50mol/ ml, CDP- ( 3a), CT P CDP-, CT P 5mol/ ml, CDP-, 50.2 % (3b )CT P, 10mol/ ml 3a C TP CD P- 3b C TP CDP- 4. Mg 2 + CDP- 4, Mg mol/ ml, CDP- ; 10mol/ ml, CDP-10mol/ ml 5. CDP , 6, CDP- (5 ) Mg 2 + CDP- 5 CDP- 6. CDP- 6 6, CDP-,, CDP-, 105

108 ) CDP K CTP M KM,,,,,,, Lineweaver-Burk, K CT M P mol/ L(7 ) 6 CDP- 7 1/ V 1/ [ S] Linewea ver-burk K C M T P mol/ L,, CT P () Borkenhagen, L.F. [ 1 ],,,, 100%, 8%,,,,, 1.,, CDP- 58% : CT P 5mol, 28mol, MgCl2 ph6.5t ris 50mol, 1.2g; 32, 6 10mol, 10mol, 2. C T P 5 - ( ATP, ),, 106

109 CDP- ATP, 1 Borkenhagen, L.F. et al. The Enzymatic Synthesis of Cytidine Diphosphate Choline, J. B iol. Chem., 1957,227(2 ) : Kennedy, E. P.. P hosphorylcholine-cytidyl Transferase, Methods in Enz ymology, 1962, ( 5 ) : ,,1983 : ,16 (2 ) 1985 : ,, 1977, (4 ) 1977 : A TP , ( 11 ) :

110 D D NA 5-5 -dam P 5 -dgmp 5 -dcm P 5 -dt MP 5 -, [ 1 ] Volkin [ 2 ] DN A 5 - Dowex-1, Dowex-2, Amberlite- IRA ,, ,,, D293 (50 ), 1. (1 ) DN A (2 ) D293 D255 JK (3 ) : 5 -dam P 5 -dgmp 5 -dcmp 5 -dtm P 2. (1 ) 5 - : [ 3 ], ph mol/ L : 40V/ cm, 5min SU V-1, 5 -, ph2.0 5mL, 2min;, 751G OD2 60 nm (2 ) : 5 ml, 30mL S HZ-82 2h, 5 -, 0.4mol/ L -0.1mol/ L 50mL, 1 2 h, OD26 0n m, 108

111 = = (OD260 nm - OD2 60 nm )30 [ OD260 nm/ g ] ( g) (OD260 nm - OD2 60 nm )30 [ OD260 nm/ ml ] 5mL OD2 60 nm 50 [OD2 60 ( g) nm/ g ] OD2 60 nm 50 [OD2 60 nm/ ml ] 5mL (3 ) D293 : D293 30mL 301.5cm,,,, ( 1), ( SV) = = ( ml/ h) ( ml) OD2 60 nm ( ml) ( ml) (4 ) 5 - : % = OD2 60 nm ( ml) OD2 60 nm ( ml) 100 [ OD260 nm/ ml ] 1., ( 2), ( OD 260nm / g ) ( OD 260nm / g ) D D J K , 1h 2 h,, 2h, D293 D255, D293 D255,, D

112 D (1 ) ph : p H, (2 ), 2 2 ph D293 ph ( OD 260nm / ml ) ( OD 260nm / ml ) p H89, p H8.5,, p H,, ph8.5 (2 ) : D293, p H 8.5, (2 )( 0.4mol/ L 0.1mol/ L ), 3 3 D293 ( OD 260nm / ml ) ( OD 260nm / ml ) , ; 5 -, p H3.5, p H ;,, Cl -, -( ) (3 ) :, D293,, 4 4 ( mol/ L ) ( O D 260nm / ml )

113 ,, 0.8mol/ L,, mol/ L (4 ) : (3 ) ( h - 1 ), SV 0.5 1, 2 000OD260 nm/ ml ; SV = 2, 1486OD260 nm/ ml ; SV, SV = 1, 2 000OD260 nm/ ml 3. D293 (1 ), 12 1 (2 ) 3 2 (3 ) :, 83%, damp dgmp dcmp dt MP 40% 111

114 5-1. OD250/ OD2 60 OD2 80/ OD26 0 OD2 90/ OD260, 2., ( 4) 4 5 -, 5-112

115 .,, 1984,119,, 2 Volkin E et al. J Am Chem Soc 1951, 73: ,16(2 ) :

116 ,,,, 732,,, 769,,, 60%,, 40 %, ( H D0011 H D0016 ),,, H D0011, H D0016, 85% 90%, 60% 710g/ L : p H8.5, 0.05 mol/ L, 300V 5, TGL16, : ( r/ min ) ( min ) 3, , , , ,000 10, 4 000r/ min r/ min,, r/ min 15,, 4, DLL3, 3 000r/ min 30,,,,, 114,,

117 P AM,,,,, 10 % 1 200mL ( 9g/ L), p H , HD0011 H D0016 H + ( 150mL ), ph3.0, ph ,, p H HD , H D , mol/ L NaO H 30, 0.1mol/ L N ao H,,,, AR, 60, 6.7g, 62%,

118 ( ), [ 1 ],, ph,,,,,, () 1. ph mol/ L ; () : ( ) ; ( ) () 2. DY-1 ( ) ; SU V-1 ( ) ; 751G () ; TGL-16 ( ) ; 10L 50L ; 25 mm75 mm 100mm75mm 116

119 ) 1. 1mL, p H mol/ L,, TGL r/ min 3min,, g, 50mL,, 50mL p H mol/ L ( 60),, 0.6 %, 60, 4mL 25 mm75mm, 20min,, 3. (2cm ),, 5 10L, ph mol/ L,,,, 300V, 5min,, 4. SU V-1 ( 1),,, 5mL p H3.0, 2min,, 751G mm O.D. 1 () 1,,, 117

120 ) 1, 2,,,, 1 C( mg) O.D nm O.D = C * 2 () 2,, 0.6% 3% 2 O.D nm

121 ) 3 3 (g ) (g) (g) ( % ) (),, ;,, 1 E., J.N., :.,1963:295,

122 5 - ( ) : -, 0.05mol/ L, 0.022mol/ L,0.012mol/ L MgSO4 7 H2 O, 0.2mol/ L, 0.2mol/ L ( ph 7.0),354, 5-42 %, 50mL, 5,5-37% : 5-5 -( ATP), ATP [ 1 ],1 mol 2mol CO2, 2molATP, ATP,, 5 - ( AM P ), ATP -A T P 1 1.1, ; -, ; AMP ( 94.6% ), ; CP : 1g -, 23mL, 1, 60, 45, 35 16g,, 0.3mol/ L KCl 2, 23mm, 0.3mol/ L KCl, ATP : ATP : ( 1 ), ph3.6, 0.05mol/ L -, 240V, 5, [ 2 ] A T P ( % ) = 120 OD260 (A T P ) OD260 ( ATP + ADP + AMP + AR) 1 A TP

123 : 50mL 16g ( ), 20mL ( 0.05mol/ L, 0.022mol/ L AMP, 0.008mol/ L MgSO4 7 H2 O, 0.2mol/ L KCl, p H7.5, 0.2mol/ L K H2 PO4 -K2 H PO4 ), 4,,35,, ATP ATP 1g,, 2.5% ( w/ w), ( 1.2.3), ATP, 1, 16g, ATP (35% ) 1 ( A mount of cells ent ra pped )/ g ( Physiological sa lin e)/ ml ( Conve rsion time)/ h Condi ti on of e ntrap ment (Conv er sion ra t e of A TP )/ % good good b ad b ad ph : ( ), p H ( 0.2mol/ L K H2 PO4 K2 H PO4 p H6.0,6.5,7.0,7.5,8.0 ), ph ATP, ph7.0 ( 42% ), 2 2 ph A TP 3 KCl A TP KCl ATP : (1.2.3 ), KCl ( KCl, 0, 0.05, 0.1, 0.2, 0.3, 0.5mol/ L ), 4h, ATP, KCl 0.2mol/ L, 42 % (3 ) Mg 2 + : (1.2.3 ), MgSO4 121

124 H2 O, 4 ATP, MgSO4 7H 2 O mol/ L, 42% (4 ) 4 MgS 4 7 H 2 O 5 AM P A T P AM P : AMP AMP 0.011, 0.022, 0.033, 0.044, 0.055mol/ L,, ( ), 4 ATP, AM P 0.022mol/ L, (5 ) : 0.05, 0.15, 0.25, 0.35mol/ L, 4, 0.05mol/ L, ATP, 42% (6 ) 6 A T P : 32, 35, 38, 41 4, 35, ATP (7) :, 0.05, 0.1, 0.2, 0.4, 0.6mol/ L ( p H7.0)4, : 20mL ( 0.05mol/ L ; 0.022mol/ L AMP; mol/ L MgSO4 7H 2 O; 0.2 mol/ L KCl; ph7.0, 0.2mol/ L ), 35, 9, 4 ATP 42% 122

125 ( C 3 - PO 3 ) A TP 9 ATP :, 50 ml, 5, ATP 37% (10 ) 10 A T P 1,.,1983, ( 11 ) :2935 2,.,1985, ( 2) :

126 ( ) ( A CDP- ATP S - ),,,,,,, CDP- 5 -, 30% 40% 60%, 1.,,, ,,, 769,,, 60 %,, 40%, ( HD0016 ), 85%,, 62% 72 % 1/ 2 2. CDP- CDP-,, 25, 35%, D H846,,,, 10,, 60% , (DN A), 717,, D H846, 124

127 90%,, 4. ( RN A), CR AR 717 UR,,

128 (, ) DEAE- DEAE-Sephadex A-50,,, 30.4U/ g, 70% ;,, 37 10, 95%, 135d ( Alkaline phospha tase, E. C , AK P ),,,,,,,, ( AM P ) 94.6%, ; DEAE-Sephadex A-50 ;, ; 1.,,, p H4.9,, ph7.5, , [ 1 ] 37.05U/ ml 2. [2 ] mol/ L, p H7.6, Tris- HCl DEAE-Sephadex A-50, 80mL, 2h, 1% 10min, 4. 1mol/ L NaCl-10-3 mol/ L MgSO mol/ L ZnSO4, 4, 40d 5., 20,, 15min 1mL, 1mL,, U/ g 126

129 (1 ) : Pi ( 2) : 20mm200mm, 50mL, 37, 37 (1%, 10-2 mol/ L MgSO4, p H9.5 ), 3mL/ min,, (3 ) : [ 2 ] ( Pi) ( ),, = ( g/ L) % 3110 :327, 31, 10 5, 30.4U/ g, U, 1 ( Table 1 ) Table 1 The activity recovery of immobilized alkaline phosphatase F ree e nzyme( ml ) T ot al activity of free en zyme( U ) T ot al activity of immobilized e nzyme( U ) T h e recov ery of th e e nzy me ac tivity( % ) ,,, 80mL, 70 % 6,,, 1. 6, 1.0, 2.0, 3.0, 4.0, 5.0mL/ min, 1 ( Fig.1 ), 3.0mL/ min ( SV3 ), 127

130 Fig T he effec t of flow rat e on th e conv er - sion ra te( K) of desoxynucleoside Fig. 2 T he effect of t emp era ture ( T ) on the conve rsion r a te ( K ) of desoxynucle o- side 2. 20, 30, 40, 45, 50, 60, 3.2mL/ min, 6, 2 ( Fig.2 ), 45,, ph p H 5.5, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0 ( 6 ), 3 ( Fig.3) ph710, 100%,, ph, p H, ph9.5 Fig. 3 T he effec t of ph on th e conver sion ra t e( K ) of desoxynu cleosid e Fig. 4 T h e effect of Mg 2 + concent ra tion( C Mg 2 + ) on th e conv er sion ra t e( K ) of d esoxynu cleosid e 4. Mg 2 + Mg 2 +, 6, 4 ( Fig.4), Mg mmol/ L,, 6.0mmol/ L, 100% Mg 2 +,,, 10mmol/ L Mg

131 1% 37, p H9.5, Mg 2 + = 10mmol/ L, 3mL/ min,, 10d, 95% (5 ( Fig.5) ) [ 3], 135d Fig. 5 T he st ability of imm obilize d alkalin e phospha t ase 1..,1983, (1 ) : ,,192. : 3, ,,377. :