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木瓜蛋白酶的原位固定化及理化性质研究(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2014年06期
Page:
99-104
Column:
研究论文
publishdate:
2014-12-09

Article Info:/Info

Title:
Study on in situ immobilization and characterization of papain
Article ID:
1000-2006(2014)06-0099-06
Author(s):
SU Erzheng1 LI Mingliang2 WEI Dongzhi2
1. Enzyme and Fermentation Technology Laboratory, College of Light Industry Science and Engineering, Nanjing Forestry University, Nanjing 210037,China;
2. State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
Keywords:
papain situ immoblization monoclonal antibody Fab fragment Fc fragment
Classification number :
TQ925
DOI:
10.3969/j.issn.1000-2006.2014.06.019
Document Code:
A
Abstract:
Amino carriers could immobilize the papain in situ in the PEG phase, which gave high papain immobilization yield(95.9%)and activity recovery(51.3%), and thus solved the difficulty in separation of the target protein and polymer after aqueous two-phase purification. The optimum pH of the immobilized papain was around 7.0, and the optimum temperature was 60-70 ℃. Compared with free enzyme, the immobilized papain was fairly stable in a wide pH(3-9)range, and the thermal stability was also improved. The best way to store immobilized papain was in wet at 4 ℃ and airtight container. The LH-HA immobilized papain could catalyze the hydrolysis of monoclonal antibody IgG to produce Fab and Fc fragments. The immobilized papain prepared in this work could be used in the production of antibody fragment industrially.

References

[1] Mitchel R E J, Chaiken I M, Smith E L. The complete amino acid sequence of papain [J]. Journal of Biological Chemistry, 1970, 245(14): 3485-3492.
[2] Kamphuis I G, Kalk K H, Swarte M B A, et al. Structure of papain refined at 1.65 Å resolution [J]. Journal of Molecular Biology, 1984, 179(2): 233-256.
[3] Kang C K, Warner W D. Tenderization of meat with papaya latex proteases [J]. Journal of Food Science, 1974, 39(4): 812-818.
[4] Starley I F, Mohammed P, Schneider G, et al. The treatment of paediatric burns using topical papaya [J]. Burns, 1999, 25(7): 636-639.
[5] Morcelle S R, Barberis S, Priolo N, et al. Comparative behaviour of proteinases from the latex of Carica papaya and Funastrum clausum as catalysts for the synthesis of Z-Ala-Phe-OMe [J]. Journal of Molecular Catalysis B: Enzymatic, 2006, 41(3): 117-124.
[6] 唐思远, 林东强, 童红飞, 等.人免疫球蛋白G酶解制备Fab和Fc片段及其分离纯化 [J].化工学报, 2013, 64(5): 1750-1756. Tang S Y, Lin D Q, Tong H F, et al. Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion[J]. CIESC Journal,2013, 64(5): 1750-1756.
[7] Adamczyk M, Gebler J C, Wu J. Papain digestion of different mouse IgG subclasses as studied by electrospray mass spectrometry [J]. Journal of Immunological Methods, 2000, 237(1-2): 95-104.
[8] Azarkan M, El Moussaoui A, van Wuytswinkel D, et al. Fractionation and purification of the enzymes stored in the latex of Carica papaya [J]. Journal of Chromatography B, 2003, 790(1-2): 229-238.
[9] Dekeyser P M, De Smedt S, Demeester J, et al. Fractionation and purification of the thiol proteinases from papaya latex [J]. Journal of Chromatography B: Biomedical Sciences and Applications, 1994, 656(1): 203-208.
[10] Kimmel J R, Smith E L. Crystalline papain. I. preparation, specificity, and activation [J]. Journal of Biological Chemistry, 1954, 207(2): 515-531.
[11] Li M L, Su E Z, You P Y, et al. Purification and in situ immobilization of papain with aqueous two-phase system[J]. PLoS ONE, 2010, 5(12): e15168.
[12] Dekeyser P M, De Smedt S, Demeester J, et al. Fractionation and purification of the thiol proteinases from papaya latex [J]. Journal of Chromatography B: Biomedical Sciences and Applications, 1994, 656(1): 203-208.
[13] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding [J]. Analytic Biochemistry, 1976, 72(1-2):248-254.

Last Update: 2014-12-31