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Purification and partial characterization of intact and truncated chitinase from Bacillus thuringiensis HZP7 expressed in Escherichia coli

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Abstract

Objective

To ascertain the effect of chitin-binding domain (ChBD) and fibronectin type III domain (FN3) on the characterization of the intact chitinase from Bacillus thuringiensis.

Results

An intact chitinase gene (chi74) from B. thuringiensis HZP7 and its truncated genes (chi54, chi63 and chi66) were expressed in Escherichia coli BL21. The expression products were analyzed after purification. All chitinases were active from pH 4–7.5 and from 20 to 80 °C with identical optimal: pH 5.5 and 60 °C. The activity of colloid chitin degradation for Chi74 was the highest, followed by Chi66, Chi63 and Chi54. Ag+ reduced the activity of Chi74, Chi54, Chi63 and Chi66, but Mg2+ enhanced them. The effect of Ag+ and Mg2+ was more significant on the activity of Chi54 than on the activities of Chi63, Chi66 and Chi74.

Conclusion

ChBDChi74 and FN3Chi74 domains play a role in exerting enzymatic activity and can improve the stability of chitinase.

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Acknowledgments

We thank Dr. Brian McGarvey for revising this manuscript. This work was supported by grants from Ministry of Science and Technology of the People’s Republic of China (No. 2011AA10A203), State Administration of Grain (No. 201313002-3) and Natural Science foundation of Fujian Province (No. 2013J01079).

Supporting information

Supplementary Table 1—Characteristics of primers.

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Authors and Affiliations

  1. Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, 350002, People’s Republic of China

    Li Sha, Ensi Shao, Xiong Guan & Zhipeng Huang

  2. School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, People’s Republic of China

    Li Sha & Zhipeng Huang

  3. National Engineering Research Center of Juncao Technology, Fujian Agriculture and Forestry University, Fuzhou, 350002, People’s Republic of China

    Ensi Shao

Authors
  1. Li Sha
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  2. Ensi Shao
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Corresponding author

Correspondence to Zhipeng Huang.

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Sha, L., Shao, E., Guan, X. et al. Purification and partial characterization of intact and truncated chitinase from Bacillus thuringiensis HZP7 expressed in Escherichia coli . Biotechnol Lett 38, 279–284 (2016). https://doi.org/10.1007/s10529-015-1970-6

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  • DOI: https://doi.org/10.1007/s10529-015-1970-6

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