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Applied Biochemistry and Biotechnology

, Volume 177, Issue 1, pp 253–266 | Cite as

Microbial Transglutaminase Separation by pH-Responsive Affinity Precipitation with Crocein Orange G as the Ligand

  • Zhaoyang Ding
  • Sipeng Li
  • Xuejun CaoEmail author
Article

Abstract

A microbial transglutaminase (MTG) was efficiently purified by using pH-responsive affinity precipitation with Crocein orange G (COG) as the affinity ligand. The docking method was used to identify the appropriate ligand. The molecular simulation results were compared with the label-free detection data analyzed by ForteBio’s Octet. A pH-responsive polymer, PMMDN, was polymerized and subsequently coupled with COG as the ligand. The isoelectric point (pI) and recovery of PMMDN and PMMDN–COG were 4.51, 99.8,% and 4.78, 98.1 %, respectively. The optimal adsorption conditions were found to be a ligand density of 60.0 μmol/g, pH 7.0, and 0.2 mol/L NaCl. The adsorption isotherm showed that the maximum adsorption capacity was 91.32 mg/g polymer and the dissociation constant was 0.021 mg/mL. Interaction information between PMMDN–COG and MTG in the whole process of affinity precipitation were obtained by Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM). PB (0.02 mol/L pH 7.0) in 20.0 % glycol was used to elute the binding MTG from PMMDN–COG. Under these conditions, electrophoretically pure MTG was obtained in only one step with elution recoveries of 98.96 % (protein) and 95.09 % (activity).

Keywords

Microbial transglutaminase Affinity precipitation pH responsive Docking Label-free detection 

Notes

Glossary

AM1-BCC

Semi-empirical (AM1) with bond charge correction (BCC)

ANTECHAMBER

Antechamber is a set of auxiliary programs for molecular mechanical studies

DMS

DMS is an open-source program written in C for computing the molecular surface of a molecule

GRID

Grid creates the grid files necessary for rapid score evaluation in DOCK

Mol2

The Mol2 file format is a popular method for specifying chemical structures, including atom types, positions, and bonding

SHOWBOX

Showbox is an interactive program for specifying the location and the size of the grids that will be calculated by the program grid

sphere_selector

Sphere_selector filters the output from sphgen selecting all spheres within a user-defined radius of a target molecule

SPHGEN

Sphgen generates sets of overlapping spheres to describe the shape of a molecule or molecular surface

sphgen_cpp

Sphgen_cpp has the same function as Sphgen

References

  1. 1.
    Ando, H., Adachi, M., Umeda, K., Matsuura, A., Nonaka, M., Uchio, R., Tanaka, H., & Motoki, M. (1989). Purification and characteristics of a novel transglutaminase derived from microorganisms. Agricultural and Biological Chemistry, 53, 2613–2617.CrossRefGoogle Scholar
  2. 2.
    Bourneow, C. and Benjakul, S. (2011) Purification and characterization of microbial transglutaminase from Enterobacter sp. C2361. Thai Journal of Agricultural Science, 496–504. Agricultural Science Society of Thailand.Google Scholar
  3. 3.
    Brozell, S., Mukherjee, S., Balius, T., Roe, D., Case, D., & Rizzo, R. (2012). Evaluation of DOCK 6 as a pose generation and database enrichment tool. J Computer Aided Molecular Design, 26, 749–773.CrossRefGoogle Scholar
  4. 4.
    Chang, R. (2000) Physical chemistry for the chemical and biological sciences.ed. University Science Books.Google Scholar
  5. 5.
    Costioli, M. D., Fisch, I., Garret-Flaudy, F., Hilbrig, F., & Freitag, R. (2003). DNA purification by triple-helix affinity precipitation. Biotechnology and Bioengineering, 81, 535–545.CrossRefGoogle Scholar
  6. 6.
    Cuatrecasas, P., & Anfinsen, C. B. (1971). Affinity chromatography. Annual Review of Biochemistry, 40, 259–278.CrossRefGoogle Scholar
  7. 7.
    Denizli, A., & Pişkin, E. (2001). Dye-ligand affinity systems. Journal of Biochemical Biophysical Methods, 49, 391–416.CrossRefGoogle Scholar
  8. 8.
    Ding, Z., & Cao, X. (2013). Affinity precipitation of cellulase using pH-response polymer with Cibacron Blue F3GA. Separation and Purification Technology, 102, 136–141.CrossRefGoogle Scholar
  9. 9.
    Ding, Z., Kang, L., & Cao, X. (2014). Application of docking methods for metal chelate affinity precipitation of endo-glucanase using pH-response polymer. Colloids and Surfaces B: Biointerfaces, 113, 412–420.CrossRefGoogle Scholar
  10. 10.
    Ding, Z., Zheng, K., & Cao, X. (2014). Lipase purification by affinity precipitation with a thermo-responsive polymer immobilized Cibacron Blue F3GA ligand. Biotechnology and Bioprocess Engineering, 19, 892–899.CrossRefGoogle Scholar
  11. 11.
    El-Hofi, M., Ismail, A., & Maher Nour, O. I. (2014). Isolation, purification and characterization of transglutaminase from rosemary (Rosmarinus officinalis l.) Leaves. Acta Scientairum Polonorum, Technologia Alimentaria, 13, 267–278.CrossRefGoogle Scholar
  12. 12.
    Ewing, T. J. A., & Kuntz, I. D. (1997). Critical evaluation of search algorithms for automated molecular docking and database screening. Journal of Computational Chemistry, 18, 1175–1189.CrossRefGoogle Scholar
  13. 13.
    Gerber, U., Jucknischke, U., Putzien, S., & Fuchsbauer, H.-L. (1994). A rapid and simple method for the purification of transglutaminase from Streptoverticillium mobaraense. The Biochemical Journal, 299, 825.CrossRefGoogle Scholar
  14. 14.
    Grossowicz, N., Wainfan, E., Borek, E., & Waelsch, H. (1950). The enzymatic formation of hydroxamic acids from glutamine and asparagine. The Journal of Biological Chemistry, 187, 111–125.Google Scholar
  15. 15.
    Gupta, M. N., Kaul, R., Guoqiang, D., Dissing, U., Mattiasson, B., & Scouten, W. H. (1996). Affinity precipitation of proteins. Journal of Molecular Recognition, 9, 356–359.CrossRefGoogle Scholar
  16. 16.
    Kumar, A., Khalil, A. A. M., Galaev, I. Y., & Mattiasson, B. (2003). Metal chelate affinity precipitation: purification of (his)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide) copolymers. Enzyme and Microbial Technology, 33, 113–117.CrossRefGoogle Scholar
  17. 17.
    Kuntz, I. D., Blaney, J. M., Oatley, S. J., Langridge, R., & Ferrin, T. E. (1982). A geometric approach to macromolecule-ligand interactions. Journal of Molecular Biology, 161, 269–288.CrossRefGoogle Scholar
  18. 18.
    Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.CrossRefGoogle Scholar
  19. 19.
    Lang, P. T., Brozell, S. R., Mukherjee, S., Pettersen, E. F., Meng, E. C., Thomas, V., Rizzo, R. C., Case, D. A., James, T. L., & Kuntz, I. D. (2009). DOCK 6: combining techniques to model RNA-small molecule complexes. RNA (New York, N.Y.), 15, 1219–1230.CrossRefGoogle Scholar
  20. 20.
    Liu, Y., Xiao, L., Joo, K.-I., Hu, B., Fang, J., & Wang, P. (2014). In situ modulation of dendritic cells by injectable thermosensitive hydrogels for cancer vaccines in mice. Biomacromolecules, 15, 3836–3845.CrossRefGoogle Scholar
  21. 21.
    Macedo, J. A., Sette, L. D., & Sato, H. H. (2011). Purification and characterization of a new transglutaminase from Streptomyces sp. Isolated in Brazilian soil. Journal of Food Biochemistry, 35, 1361–1372.CrossRefGoogle Scholar
  22. 22.
    Memmedova, T., Armutcu, C., Uzun, L., & Denizli, A. (2015). Polyglycidyl methacrylate based immunoaffinity cryogels for insulin adsorption. Materials Science and Engineering: C, 52, 178–185.CrossRefGoogle Scholar
  23. 23.
    Meng, E. C., Shoichet, B. K., & Kuntz, I. D. (1992). Automated docking with grid-based energy evaluation. Journal of Computational Chemistry, 13, 505–524.CrossRefGoogle Scholar
  24. 24.
    Motoki, M., & Seguro, K. (1998). Transglutaminase and its use for food processing. Trends if Food Science and Technology, 9, 204–210.CrossRefGoogle Scholar
  25. 25.
    Pencheva, T., Soumana, O. S., Pajeva, I., & Miteva, M. A. (2010). Post-docking virtual screening of diverse binding pockets: comparative study using DOCK, AMMOS, X-Score and FRED scoring functions. European Journal of Medicinal Chemistry, 45, 2622–2628.CrossRefGoogle Scholar
  26. 26.
    Pettersen, E. F., Goddard, T. D., Huang, C. C., Couch, G. S., Greenblatt, D. M., Meng, E. C., & Ferrin, T. E. (2004). UCSF Chimera—a visualization system for exploratory research and analysis. Journal of Computational Chemistry, 25, 1605–1612.CrossRefGoogle Scholar
  27. 27.
    Porath, J., Carlsson, J. A. N., Olsson, I., & Belfrage, G. (1975). Metal chelate affinity chromatography, a new approach to protein fractionation. Nature, 258, 598–599.CrossRefGoogle Scholar
  28. 28.
    Sheth, R. D., Madan, B., Chen, W., & Cramer, S. M. (2013). High-throughput screening for the development of a monoclonal antibody affinity precipitation step using ELP-z stimuli responsive biopolymers. Biotechnology and Bioengineering, 110, 2664–2676.CrossRefGoogle Scholar
  29. 29.
    Shi, Y. G., Qian, L., Zhang, N., Han, C. R., Liu, Y., Zhang, Y. F., & Ma, Y. Q. (2012). Study on separation and purification of the transglutaminase. Applied Mechanics and Materials, 121, 443–447.Google Scholar
  30. 30.
    Strop, P. (2014). Versatility of microbial transglutaminase. Bioconjugate Chemistry, 25, 855–862.CrossRefGoogle Scholar
  31. 31.
    Subramanian, S. and Ross, P. D. (1984) Dye-ligand affinity chromatography: the interaction of Cibacron Blue F3GA® with proteins and enzyme. Critical Reviews Biochemistry Molecular,16, 169-205.Google Scholar
  32. 32.
    Totrov, M., & Abagyan, R. (2008). Flexible ligand docking to multiple receptor conformations: a practical alternative. Current Opinion in Structural Biology, 18, 178–184.CrossRefGoogle Scholar
  33. 33.
    Vaidya, A. A., Lele, B. S., Deshmukh, M. V., & Kulkarni, M. G. (2001). Design and evaluation of new ligands for lysozyme recovery by affinity thermoprecipitation. Chemical Engineering Science, 56, 5681–5692.CrossRefGoogle Scholar
  34. 34.
    Wilhelm, B., Meinhardt, A., & Seitz, J. (1996). Transglutaminases: purification and activity assays. Journal of Chromatography B: Biomedical Sciences and Applications, 684, 163–177.CrossRefGoogle Scholar
  35. 35.
    Yan, B., & Cao, X. (2012). Preparation of aqueous two-phase systems composed of two pH-response polymers and liquid–liquid extraction of demeclocycline. Journal of Chromatography. A, 1245, 39–45.CrossRefGoogle Scholar
  36. 36.
    Yılmaz, M., Bayramoǧlu, G., & Arıca, M. Y. (2005). Separation and purification of lysozyme by Reactive Green 19 immobilized membrane affinity chromatography. Food Chemistry, 89, 11–18.CrossRefGoogle Scholar
  37. 37.
    Yokoyama, K., Nio, N., & Kikuchi, Y. (2004). Properties and applications of microbial transglutaminase. Appl Microbiol Biot, 64, 447–454.CrossRefGoogle Scholar
  38. 38.
    Zhu, Y., & Tramper, J. (2008). Novel applications for microbial transglutaminase beyond food processing. Trends in Biotechnology, 26, 559–565.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.State Key Laboratory of Bioreactor Engineering, Department of BioengineeringEast China University of Science and TechnologyShanghaiChina

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