Please use this identifier to cite or link to this item:
Refolding of D-hydantoinase with fed-batch dilution process
|關鍵字:||D-hydantoinase;乙內醯尿酶;refolding||出版社:||化學工程學系所||引用:|| Mitraki A., J. King(1989) Protein folding intermediates and inclusion body formation , Bio/technology, 7:690-697  Xie Y., Wetlaufer D B.(1996) Control of aggregation in protein refolding:The temperature-leap tactic ,Protein Science,5(3):517~523  S. Misawa , I. Kumagai (1999) Refolding of therapeutic proteins produced in Escherichia coli as inclusion bodies, Biopolymers (Peptide Science) 51:297~307  林藹寧。生技產品之回收與純化技術，生物技術(2002)  Batas B.,Schiraldic C., Chaudhuri(1999), Inclusion body purification and protein refolding using microfiltration and size exclusion chromatography, J. of Biotechnology,68:149~158  Neurath,H.,Greenstein, J. P. ,Putnam,F. W.,Erickson J. O.(1943) The Chemistry of protein denaturation ,Chem. Rev.,32:157~265  Campbell, M.K.,（1995）“Chapter 4: The three-dimensional structure of protein”, in Biochemistry, Campbell, M.K., Saunders College Publishing.  Machida S.（2000）Cycloamylose as an efficient artifical chaperone for protein refolding. FEBS Lett. 486：131-135  Yoshimoto M.,Kuboi R.(1999) Oxidative Refolding of Denatured/Reduced Lysozyme Utilizing the Chaperone-like Function of Liposomes and Immobilized Liposome Chromatography, Biotechnol. Prog.,15:480-487  Diane L. Hevehan, Eliana De Bernardez Clark(1997) Oxidative renaturation of lysozyme at high concentration , Biotechnology and Bioengineering , 54:221~230  Batas B.,Julian B. Chaudhuri(1996) Protein Refolding at High Concentration Using Size-Exclusion Chromatography, Biotechnology and Bioengineering ,50:16~23  De Bernardez Clark E(2001) Protein refolding for industrial processes, Current Opinion in Biotechnology,12:202~207  Carpenter JF,Kendrick BS,Chang BS,Manning MC,Randolph TE (1999) Inhibition of stress-induced aggregate of protein therapeutics. Method Enzymol.,309:236~255  Matsubara M, Nohara D, Kurimoto E, Kuroda Y, Sakai T.,(1993) "Loose folding" and "delayed oxidation" procedures successfully applied for refolding of fully reduced hen egg white lysozyme. Chem Pharm Bull (Tokyo). ,41(7):1207~1210.  Katoh,S.,Terashima M.,Kishida H.,Yagi H.(1997) Refolding efficiency of lysozyme in fed-batch system,J.Chem. Eng. Jpn., 30:964~966  Katoh S, Sezai Y, Yamaguchi T, Katoh Y, Yagi H, Nohara D(1999) Refolding of enzymes in a fed-batch operation, Process Biochemistry 35 :297~300  Katoh S, Katoh Y,Continuous refolding of lysozyme with fed-batch addition of denatured protein solution,Process Biochemistry 35(2000) 1119~1124 [18 Fischer B., Perry B.,Sumner, I. , Goodenough,P.(1992) A novel sequential procedure to enhance the renaturation of recombinant protein from Escherichia coli inclusion bodies,Protein engineering,5:593~596  Altenbuchner J., M. Siemann-Herzberg, and C. Syldatk. (2001). Hydantoinasees related enzymes as biocatalysts for the synthesis of unnatural chiral amino acids. Curr Opin Biotechnol. 12：559-563.  Cheon, Y. H., H. S. Kim, K. H. Han, J. Abendroth, K. Niefind, D. Schomburg, and J. Wang. (2002). Crystal structure of D-hydantoinase from Baccilus stearothermophilus: Insight into the stereochemistry of enantioselectivity. Biochemistry. 41：9410-9417  高肇鴻。1998。Bacillus circulans D-hydantoinase 基因的選殖、表現及酵素特性。碩士論文。分子生物研究所。中興大學。台中。||摘要:||
在利用大腸桿菌生產重組乙內醯尿酶時，其胞內會形成大量不具活性的包涵體(inclusion bodies)，由於在包涵體內含有高純度的蛋白，若能有效的恢復其活性，則應用於工業上具有相當大的經濟價值。故欲發展出一個最適的復性程序，將包涵體回復成具有活性的蛋白質。實驗中先以pH 12.5, 50mM Tris-bufferru將包涵體溶解，再藉由稀釋法使其復性，並探討直接稀釋法與餽料稀釋法對乙內醯尿酶復性的影響，另外探討添加劑是否對可溶蛋白回收率有所幫助。依據實驗結果，在低流速時復性效果較佳，但添加劑對於可溶蛋白回收率並無明顯的效果。
Recombinant D-hydantoinase was over-expressed in Escherichia coli in the form of inclusion bodies, which contained large amount of the targeted proteins are valuable for industrial application if these protein could be reforded. An optimal refolding procedure has been developed to reford active proteins from inclusion bodies. The inclusion bodies were solubilized in 50mM Tris-buffer at pH 12.5 and then were diluted to refold the denatured proteins. Refolding of D-hydantoinase by direct and fed-batch operations was investigated . Furthermore, the influence of the additives, CTAB and MnCl2, for soluble protein recovery was also studied. The results of the experiments indicate that refolding process with low velocity in the fed-batch operation resulted in higher recovery of soluble protein and it is not recommended to add additives for refolding procedure.
|Appears in Collections:||化學工程學系所|
Show full item record
TAIR Related Article
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.