Erratum to “Expression, Purification and Crystallization of Thermostable Mutant of Cutinase Est1 from Thermobifida alba.” [Advances in Bioscience and Biotechnology 9 (2018), 215-223]


The original online version of this article (Kitadokoro, K., Matsui, S., Osokoshi, R., Nakata, K. and Kamitani, S. (2018) Expression, Purification and Crystallization of Thermostable Mutant of Cutinase Est1 from Thermobifida alba. Advances in Bioscience and Biotechnology, 9, 215-223. unfortunately contains some mistakes. The author wishes to correct the errors.



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Kitadokoro, K. , Matsui, S. , Osokoshi, R. , Nakata, K. , Thumarat, U. , Kawai, F. and Kamitani, S. (2018) Erratum to “Expression, Purification and Crystallization of Thermostable Mutant of Cutinase Est1 from Thermobifida alba.” [Advances in Bioscience and Biotechnology 9 (2018), 215-223]. Advances in Bioscience and Biotechnology, 9, 378-379. doi: 10.4236/abb.2018.98025.

2.1. Protein Expression and Purification

Est1(A68V/T253P) was used throughout this study (Table 1) [6]. Briefly, 20 mL of an overnight culture of E.coli cells Rosetta-gami B (DE3) transformed with pQE80L-est1(A68V/T253P) was inoculated to 400 mL of LB medium with 50 µg∙ml−1 ampicillin.

Determination of Specific Activity

Enzymatic activities were determined by using p-nitrophenylbutyrate (pNPB) ester substrates as previously described [6]. The reaction was performed at 310 K in 1 mL of the mixture containing 50 mM Tris-HCl buffer (pH 8.0), 1 mM pNPB and 0.001 mM enzyme. The reaction mixture without the enzyme was used as the control. Reactions were started by the addition of pNPB.


We are grateful to all members of beamline BL44XU at SPring-8 for their help in collecting data. Use of the synchrotron beamline BL44XU at SPring-8 was obtained through the Cooperative Research Program of the Institute for Protein Research, Osaka University.

Table 1. Macromolecule production information [6].

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Pio, T.F. and Macedo, G.A. (2009) Cutinases: Properties and Industrial Applications. Advances in Applied Microbiology, 66, 77-95.
[2] Hu, X., Thumarat, U., Zhang, X., Tang, M. and Kawai, F. (2010) Diversity of Polyester-Degrading Bacteria in Compost and Molecular Analysis of a Thermoactive Esterase from Thermobifida alba AHK119. Applied Microbiology and Biotechnology, 87, 771-779.
[3] Dresler, K., Heuvel, J., Muller, R.J. and Deckwerl, W.D. (2006) Production of a Recombinant Polyester-Cleaving Hydrolase from Thermobifida fusca in Escherichia coli. Bioprocess and Biosystems Engineering, 29, 169-183.
[4] Acero, E.H., Ribitsch, D., Steinkellner, G., Gruber, K., Greimel, K., Eiteljoerg, I., Trotscha, E., Wei, R., Zimmermann, W., Zinee, M., Cavco-Paulo, A., Freddi, G., Schwab, H. and Guebitz, G. (2011) Enzymatic Surface Hydrolysis of PET: Effect of Structural Diversity on Kinetic Properties of Cutinases from Thermobifida. Macromolecules, 44, 4632-4640.
[5] Thumarat, U., Nakamura, R., Kawabata, T., Suzuki, H. and Kawai, F. (2012) Biochemical and Genetic Analysis of a Cutinase-Type Polyesterase from a Thermophilic Thermobifida alba AHK119. Applied Microbiology and Biotechnology, 95, 419-430.
[6] Thumarat, U., Kawabata, T., Nakajima, M., Nakajima, H., Sugiyama, A., Yazaki, K., Tada, T., Waku, T., Tanaka, N. and Kawai, F. (2015) Comparison of Genetic Structures and Biochemical Properties of Tandem Cutinase-Type Polyesterases from Thermobifida alba AHK119. Journal of Bioscience and Bioengineering, 120, 491-497.
[7] Kitadokoro, K., Thumarat, U., Nakamura, R., Nishimura, K., Karatani, H., Suzuki, H. and Kawai, F. (2012) Crystal Structure of Cutinase Est119 from Thermobida alba AHK119 that Can Degrade Modified Polyethylene Terephthalate at 1.76 A Resolution. Polymer Degradation and Stability, 97, 771-775.
[8] Roth, C., Wei, R., Oeser, T., Then, J., Follner, C., Zimmermann, W. and Strater, N. (2014) Structural and Functional Studies on a Thermostable Polyethylene Terephthalate Degrading Hydrolase from Thermobifida fusca. Applied Microbiology and Biotechnology, 98, 7815-7823.
[9] Ribitsch, D., Hromic, A., Zitzenbacher, S., Zartl, B., Gamerith, C., Pellis, A., Jungbauer, A., Lyskowski, A., Steinkellner, G., Gruber, K., Tscheliessnig, R., Herrero Acero, E. and Guebitz, G.M. (2017) Small Cause, Large Effect: Structural Characterization of Cutinases from Thermobifida cellulosilytica. Biotechnology and Bioengineering, 114, 2481-2488.
[10] Jancarik, J. and Kim, S.H. (1991) Sparse Matrix Sampling: A Screening Method for Crystallization of Proteins. Journal of Applied Crystallography, 24, 409-411.
[11] Otwinowski, Z. and Minor, W. (1997) Processing of X-Ray Diffraction Data Collected in Oscillation Mode. Methods in Enzymology, 276, 307-326.
[12] Matthews, B.W. (1968) Solvent Content of Protein Crystals. Journal of Molecular Biology, 33, 491-497.
[13] Vagin, A. and Teplyakov, A. (2010) MOLREP: An Automated Program for Molecular Replacement. Acta Cryst, D66, 22-25.
[14] Winn, M.D., Ballard, C.C., Cowtan, K.D., Dodson, E.J., Emsley, P., Evans, P.R., Keegan, R.M., Krissinel, E.B., Leslie, A.G.W., McCoy, A., McNicholas, S.J., Murshudov, G.N., Pannu, N.S., Potterton, E.A., Powell, H.R., Read, R.J., Vagin, A. and Wilson, K.S. (2011) Overview of the CCP4 Suite and Current Developments. ActaCryst, D67, 235-242.

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