1、F ) Z王佃亮*(t bt?v 3“ 100083)K1 与其他来源的溶菌酶相比, 人溶菌酶具有独特的优越性和多种多样的药理作用效果, 在临床上具有多种重要应用价值b但天然人溶菌酶来源极其困难, 利用重组DNA 技术进行生产是解决这一难题的有效途径b迄今人们已利用化学合成或从人类细胞组织中制备 cDNA等途径获取人溶菌酶基因, 并在大肠杆菌a酵母菌和真菌表达系统中进行了表达, 最高水平为40mgPL,低于人乳中溶菌酶含量( 50 250mgPL) ,虽然距离工业化生产仍有一定距离b但重组人溶菌酶发展前景看好b1oM 人溶菌酶 重组人溶菌酶 DNA重组技术 药理作用 临床应用l : 2003-
2、06-18 : 2003-07-22*0Q: wangdianl sina. com ) ( human lysozyme) Ba o,130 F,s0 14600,4= ;o 1 , B) F ) vs, ) )dM , ) bWM o2b“ ) , “5v? 3 s jb1 F ) 7?A1111 ) ) “ - “5 ) 1 b ), Bs, 8 = 3 fTb ) 5 , 8 =B, 8 ? M , “5 H,1 1 ) , O OT 3,4bN, ) 1 b ) ) b ) )1 b ) 3, O 9b ) 4, 5b112 ) ,i9 b ,9 ;7M iCa2+aNi+aMn2+
3、 YlCu2+b212 8 T Murakami 15*5 S Vr ) y,Vr d8 Ti, 3b Yoshimura 16YV ) y B ) |t,V7 P S )s ) ,Vr 515mgPLbKI_ 17| )y X H 8 ( Pichia pastoris)sVr8pPIC9K,yF pPIC9K-hLY,Bgl ML, dE| 8 % =,YVG418 , X HJ?Vr,KVr ! b90147, 1Vr 5 4b213 )T Andrew 18 ) w ( Aspergillus niger) 3F ) H,9 “d 3F ) V ?b ) cDNA PCR( recur
4、sive PCR),$ X HVr8pIGF,“ w , Y 6( PVP) ! !,K 40mgPLb7M !Hq/,F ) V50mgPLbShigeru 3 ) y ( Acremonium chrysogenum)Vr, y 4Vr8,1 uYa 0 ) yW(V1) , pNLH2Vr K,V 40mgPml, ) y F a|t-t yVrbHideyuki 199,YVDNAF/ , ) FBavl t,9F ) f| ) )rTb60 S 3 23 V1 ) Vr8 Vr 1 uY% ) ( mgPL) (KvpNLH1 ,a |t, 1113 2312pNLH2 ,a |t)
5、a -t, 3612 4613pNLH3 ,a |t)a -t) t, 1917 2515pNLH4 , ) |t, 1110 15133 i5# % ) Bs0 v,% ) ?rVr, 7 L= f X, y V ?+Z bB,yVrq( 0a9 , u. ) # J . J ?, 1995, ( 5): 80 82 5 = +, + W *. ) ) B. 3,1999, 39( 1) : 55 59 6 k, 1 . ) Z.S 30,1999, 20( 6) : 319 320 7 L :, . ) Z.S ? “5, 1994,21( 9) : 709 711 8 Hisham R
6、I, Tetsuji M , Takayoshi A, et al. Genetic evidence thatantibacterial activity of lysozyme is independent of its catalyticfunction. FEBS Letters, 2001, (506) : 27 32 9 Pepys M B, Hawkins P N, Booth D R, et al. Human lysozyme genemutations cause hereditary systemic amyloidosis. Nature, 1993, 362:553
7、557 10 Caballero M, Ruiz R, Marquez M, et al. Development of aM icroparticle-Enhanced Nephelometric Immunoassay forQuantitation of Human Lysozyme in Pleural Effusion and Plasma.Journal of Clinical Laboratory Analysis, 1999, ( 13) : 301 307 11=, gk *, . b ) 4 |# .? , 1996, (1) : 37 4012 + W *, +,=,.V
8、v )c84 | ) . 3, 1996, 12(9 ) : 266 26813 i, + W (,= +,. ) yv)Vr. 3, 1997, 13( 1) : 102 10414. F# ?.SD rs,2002, 13( 6) : 243 24515 Murakami M, Jigami Y, Tanaka H, et al. Expression of synthetichuman lysozyme gene in Escherichia coli. Agricultural BiologicalChemistry, 1986, ( 50) : 713 72316 Yoshimura
9、 K, Toibana A, Nakahama K. Human lysozyme: Sequenceof a cDNA, and expression and secretion by Saccharomycescerevisae. Biochemistry and Biophysics Research Communication,1988, ( 105) : 794 80117I_,q, ,. ) y X H# 8 Vr. +Dv, 2001, 22 ( 22) : 2068 207218 Andrew S, Ludmilla A M, Wim N, et al. Expression,
10、 purification,and characterization of the recombinant calcium-binding equinelysozyme secreted by the filamentous fungi Aspergillus niger :Comparisions with the production of hen and human lysozymes.Protein Expression and Purification, 1999, ( 16) : 171 18019 Hideyuki A, Hisham R I, Takeshi K, et al.
11、 Bactericidal action oflysozymes attached with various sizes of hydrophobic peptides to theC-terminal using genetic modification. FEBS Letters, 1997, 415: 114 11820k, ,. ) Z. 3,2002, 32( 4) : 55 5821 . ) # J . J , 1999, 3: 212262 S 3 23 Research Progress in Recombinant Human LysozymeWang Dianliang(
12、Department of Biotechnology, Beijing University of Aeronautics and Astronautics Beijing 100083)Abstract Compares with other types of lysozymes, human lysozyme possesses many particular advantages andpharmacological effects, therefore, it will be widely used in clinical medicine field. However, for r
13、easons that thebiomaterials from which authentic human lysozyme is fabricated are quite short, the production of recombinant humanlysozyme by means of genetically-engineered techniques will become an effective method to solve this problem. Humanlysozyme gene was so far made from chemical synthesis p
14、athway, and human lysozyme cDNA from human cell or tissueas well. All these genes were transferred into E . coli, yeast and filamentous fungi respectively, and a maximum humanlysozyme expression of 40mgPL was obtained when A. chrysogenum, a kind of filamentous fungi, was used as host. Theexpression
15、level was quite low while human milk contains 50mgPL 250mgPL. Although human lysozyme was by nowimpossible to produce on large-scale, it definitely have a prospective future.Key words Human lyszyme Recombinant human lysozyme Rrecombinant DNA technique Pharmacologicaleffect Clinical application2002 M
16、5/ Y6()()5/ Y6()5/ Y6() SE S/ ,SE/ ?Z9( 8639) iS S/ 8 , S/ # H?V TS =aS= ,S/ ?Z v S =Yb ,v16 7, 112:, |ISSN1002-0470aCN11-2770PN;1 ,v167, 96:, |ISSN1006-6748aCN11-3683PNb = #Ya 3C ja15/a?a a/ 5,1 S a S/5 avn =3#v S b 1+ = , yb/Y0 B +w 7 / T, S = n7, W5 b l Xa aZE/ , a La V b N15100(c) , N25100(c) , Z T:,: g2143Q5/ Y6I ( 100045) , “ :,: g g s ),k|: 0200004609014463033, : S/ D( h 5/ Y6 n) ,I :( 010) 68511133-1272, 68514060, Fax: ( 010) 68511839, Email: hitech istic. ac. cn639 :F ) Z
