V o l. 19 N o. 10 1 9 9 8 10 CH EM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES 1584 1588 (g ) (,,, 300071) (, ),,..,, 1. 0 g1,., 33. 1%.,,, O 632. 5, O 629. 7. (H b),., Bonaven tu ra 80, H b, H b 50 100 m g, 60% 80% [1 3 ]. A quanau tics,, [4, 5 ]..,, H b,,.,,. 1 1. 1 ( ) ; ( ) ; ( ) ; ( ) ; ( ) ; ( ) ; ( ) ; (, ). TH Z282 ; 180280 ; H itach i 20PR 2520. 1. 2 1. 2. 1 [ 6 ], 4, 5 000 rgm in 10 m in, : 1997211204. :,, 31,,.
N o. 10 : (g ) 1585, 2., 4 30 60 m in, 0. 4, 15 m in, 4, 7 000 rgm in 10 m in, H b. N 2 (0. 3 Λm ), 4. H b, H b. [H b ] = [Fe ] 64500 = 287. 9[Fe ] 56 4 1. 2. 2 1. 0 g 20 ml 0125 mo lgl N a IO 4, 30 3 h, N a IO 4. 25 ml 5 mo lgl N aoh, 70 30 m in,,. 30 20 ml 0125 mo lgl N a IO 4 6 h, N a IO 4. [ 7 ],. 011 g, 20 ml 013 mo lgl, 15 0104%, 70 2 h,, 0103 mo lgl N aoh ( ).,. 1. 2. 3 H b H b, 4,, F ig. 1 Sketch of the gas volumetr ic apparatus H b,, 1. M agnetic stirrer; 2. T emperature2constant H b, H b bath; 3. F lask; 4. Inlet; 5. F ine gas line;. 6. 5 ml P ipet; 7. L eveling bulb. 1. 2. 4 ( 1) [ 8 ], 011.,, K 3Fe (CN ) 6,,,., H b ( H b 1. 39 ml ). 2 2. 1 1010 g 200 g 20% N aoh, 8 h,, 50. ( 0125 mo lgl N a IO 4, 30 ), 1.,., 1,, O xidation tim e Table 1 Effect of alkaline treatmen t on cellulose ox idation Amount of aldehyde groupg(mmo l g - 1 ) gh O riginal cellulo se A lkaline treated cellulo se 2 1. 85 4. 08 6 4. 00 6. 75 12 5. 73 9. 50 24 7. 11 10101,,..
1586 V o l. 19 2. 2 Hb 2. 2. 1 H b H b 4, H b, H b.,,, H b ( 2),.,,, H b. Table 2 The amoun t of immobilized Hb (Hb concen tration: 11010 ggl ) on simply ox idized cellulose Amount of aldehyde groupg(mmo l g - 1 ) 1. 85 4. 00 5. 73 7. 11 Amount of immobilized H bg(m g g - 1 ) 11. 2 25. 3 36. 7 42. 6 2. 2. 2 H b 3, H b. 1 2,, H b. 3,,,,, H b 223. 5 m g g.,, H b 784. 5 m g g. Table 3 The amoun t of Hb immobilized on cellulose vs the treatmen t procedure of cellulose 3 (Hb concen tration: 11010 ggl ) Amount of aldehyde group Status of Amount of immobilized H b T reatm ent p rocedure g(mmo l g - 1 ) oxycellulo se g(m g g - 1 ) 1. O xidation 2. 4 W h ite co tton 102. 3 2. A lkaline treatm ent; oxidation 4. 6 W h ite co tton 151. 5 3. O xidation; alkaline treatm ent 0 Gel 223. 5 4. P re2oxidation; alkaline treatm ent; 1. 76 Gel 784. 5 O xidation 3 Reaction conditions fo r oxidation: 0125 mo lgl N a IO 4, 30, 4 h; p re2oxidation: 0125 mo lgl N a IO 4, 30, 3 h; alkaline treatm ent: 5 mo lgl N aoh, 70, 30 m in. 2. 2. 3 H b 3,,. M eara [9, 10 ], Β C O, Cann izzaro,., H b.,.,, H b., H b,,, H b ( 4). 1 0125 mo lgl N a IO 4, 30, 3 h. Table 4 Effect of pre-ox idation on Hb immobilization 3, P re2oxidation tim egh 015 1 2 3 4 6 12 Yield of final p roduct (% ) 57. 7 44. 8 35. 3 32. 2 31. 5 25. 5 0 Amount of immobilized H b (m g g - 1 ) 479. 7 751. 5 1 00019 1 141. 9 1 084. 2 657. 2 0 3 T he second oxidation: 30, 6 h; H b concentration: 11010 ggl. 2. 2. 4 H b,, H b,, H b. 0125 mo lgl N a IO 4, 30, 3 h,, 0125 mo lgl N a IO 4 30 1 5,,, H b.,,, 6 h.
N o. 10 : (g ) 1587 Table 5 Effect of the second ox idation on Hb immobilization (Hb concen tration: 116. 0 ggl ) Second oxidation tim egh 2 4 6 8 Yield of oxycellulo se (% ) 36. 8 34. 2 28. 6 2010 Amount of aldehyde groupg(mmo l g - 1 ) 2. 09 2. 22 4. 03 4. 03 Amount of immobilized H bg(m g g - 1 ) 652. 2 853. 8 1 146. 9 1 20010 2. 3 3. 0 g (0125 mo lgl N a IO 4, 30, 3 h; 5 mo lgl N aoh, 70, 30 m in; 0125 mo lgl N a IO 4, 30, 6 h), 40 ml H b (133. 9 ggl ), 4,, 1015 ml, H b 1 025. 2 g., H b. K 3Fe (CN ) 6 ( 2), 33. 1%., H b, H b F ig. 2 Oxygen release of oxygen carr ier based on Hb linked per iodate oxy2, H b, cellulose. 1 Celia Bonaventura, Jo seph Bonaventura, Irving R. Hooper et al.. A pp l. B iochem. B io techno l., 1984, 9: 65 2 Jo seph Bonaventura, Celia Bonaventura. U. S. Patent, 4 343 715, 1982 3 Jo seph Bonaventura, Celia Bonaventura. U. S. Patent, 4 602 987, 1986 4 D e Castro E. S., Benedict C., Caro llo V. et a l.. ECS Sympo sium, 1989 5 D e Castro E. S.. Chem tech., 1990, 20 (11) : 682 6 D e V enuto F., Zuck T. F., Zegna A. İ et a l.. J. L ab. C lin. M ed., 1977, 89 (3) : 509 7 M aute R. L., Ow ens M. L.. A nal. Chem., 1965, 28: 1 312 8 D ian Chen, M artell A. E.. Ino rg. Chem., 1987, 26: 1 026 9 R ichards G. N., Seph ton H. H.. J. Chem. Soc., 1957: 4 492 10 M eara D. O., R ichards G. N.. J. Chem. Soc., 1958: 4 504 Oxygen Carr ier Based on Imm obilized Hem oglobin (g ) Hem oglobin Cova len tly Bond to Per ioda te Oxycellulose KON G D e2l ing 3, J IA Yong2H u i, YU Yao2T ing, TON G M ing2rong (Institu te f or M olecu lar B iology, B ioactive M aterials R esearch L aboratory, N ankai U niversity, T ianj in, 300071) W ON G T ze2fei (D ep artm ent of B iochem istry, H ong K ong U niversity of S cience and T echnology, H ong K ong ) Abstract Cellu lo se can be easily ox idized to oxycellu lo se w ith periodic acid, w h ich can serve
1588 V o l. 19 as a carrier fo r the immob ilization of hemoglob in (H b ) acting like a b ion ic gill, to ex tract oxygen from sea w ater, w h ich m ay p rovide a sou rce of oxygen fo r hum an life suppo rt in an undersea environm en ṫ T he ob jective of the p resen t paper is to study the po ssib ility of u sing modified cellu lo se derivatives as a carrier to immob ilize a h igh amoun t of H b. D ifferen t param eters influencing immob ilization w ere investigated. It w as found that on2 ly by the un ique activation p rocedu re, that co tton w as first ox idized w ith N a IO 4, then treat2 ed w ith 5 mo lgl N aoh and again ox idized w ith N a IO 4. T he carrier thu s ob tained w as ab le to immob ilize a very h igh amoun t of H b, i. e. 1. 0 g g cellu lo se. W hen cellu lo se w as ox idized on ly once w ith N a IO 4, although the aldehyde group con ten t w as rather h igh, the link ing amoun t of H b w as very low. T h is is p robab ly due to the m icrocrystalline structu re in cellu2 lo se. A lkaline degradation of periodate oxycellu lo se led to destruction of the m icrocrystalline structu re. T he alkaline degradation and Cann izzaro reaction also resu lted in a comp lete lo ss of aldehyde group. In o rder to ob tain m ax im um amoun t of immob ilized H b, a second ox ida2 tion w ith N a IO 4 w as necessary to p roduce aldehyde group s. A fter ox idation w ith po tassium ferricyan ide, the oxygen carrier show ed an oxygen release efficacy of 33. 1%. Keywords O xygen carrier, O xycellu lo se, Immob ilized hemoglob in, A rtificial gill (Ed. : W, L ) ( IS I) (C itation rate) (impact facto r) (imm ediacy index), ; ;,,,,,,,, IS I SC I,,, SC I SC I, 1997 SC I 6, 31,,,,,,,,,,,,,,,,,,, ( 1596 )