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Purification of a thermostable chitinase from Bacillus cereus by chitin affinity and its application in microbial community changes in soil

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Abstract

A thermostable chitinase was purified by chitin affinity from the culture supernatant of Bacillus cereus TKU028 with shrimp head powder (SHP) as the sole carbon/nitrogen source. TKU028 chitinase was purified using a one-step affinity adsorbent system, and the molecular mass of TKU028 chitinase (approximately 40 kDa) was then determined using SDS-PAGE. The enzyme was stable for 60 min at temperatures below 60 °C and stable over a broad pH range of 4–9 for 60 min. In addition, the temporal changes of a bacterial community in mangrove river sediment of the Tamsui River with added SHP were also analysed by PCR–denaturing gradient gel electrophoresis to investigate the effects of B. cereus TKU028 on the degradation of SHP. The 6-week incubation sample of SHP and B. cereus TKU028-amended mangrove river sediment displayed the highest amount of biomass, reducing sugar and total sugar, and some variance of bacterial community composition existed in the soils.

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Acknowledgments

This work was supported in part by a Grant of the National Science Council, Taiwan (NSC99-2313-B-032-001-MY3).

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Correspondence to San-Lang Wang.

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Liang, T., Hsieh, T. & Wang, S. Purification of a thermostable chitinase from Bacillus cereus by chitin affinity and its application in microbial community changes in soil. Bioprocess Biosyst Eng 37, 1201–1209 (2014). https://doi.org/10.1007/s00449-013-1092-2

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Keywords

  • Chitinase
  • Shrimp heads
  • Bacillus cereus
  • Affinity adsorption
  • PCR–DGGE