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β-cyclodextrin production by the cyclodextrin glucanotransferase from Paenibacillus illinoisensis ZY-08: cloning, purification, and properties

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Abstract

The gene encoding the cyclodextrin glucanotransferase (CGTase, EC2.4.1.19) of Paenibacillus illinoisensis was isolated, cloned, sequenced and expressed in Escherichia coli. Sequence analysis showed that the mature enzyme (684 amino acids) was preceded by a signal peptide of 34-residues. The deduced amino acid sequence of the CGTase from P. illinoisensis ZY-08 exhibited highest identity (99 %) to the CGTase sequence from Bacillus licheniformis (P14014). The four consensus regions of carbohydrate converting domain and Ca2+ binding domain could be identified in the sequence. The CGTase was purified by using cold expression vector, pCold I, and His-tag affinity chromatography. The molecular weight of the purified enzyme was about 74 kDa. The optimum temperature and pH of the enzyme were 40 °C and pH 7.4, respectively. The enzyme activity was increased by the addition of Ca2+ and inhibited by Ba2+, Cu2+, and Hg2+. The K m and V max values calculated were 0.48 mg/ml and 51.38 mg of β-cyclodextrin/ml/min. The ZY-08 and recombinant readily converted soluble starch to β-cyclodextrin but ZY-08 did not convert king oyster mushroom powder and enoki mushroom powder. However the recombinant CGTase converted king oyster mushroom powder and enoki mushroom powder to β-cyclodextrin.

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Acknowledgments

This work was supported by Dong-A University Research Fund.

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Authors and Affiliations

  1. Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University, Busan, 604-714, Republic of Korea

    Yong-Suk Lee, Dong-Ju Park, Jie Chang & Yong-Lark Choi

  2. College of Agriculture, Yangtze University, Jingzhou, Hubei Province, People’s Republic of China

    Yi Zhou

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  1. Yong-Suk Lee
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  2. Yi Zhou
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Correspondence to Yong-Lark Choi.

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Lee, YS., Zhou, Y., Park, DJ. et al. β-cyclodextrin production by the cyclodextrin glucanotransferase from Paenibacillus illinoisensis ZY-08: cloning, purification, and properties. World J Microbiol Biotechnol 29, 865–873 (2013). https://doi.org/10.1007/s11274-012-1241-9

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