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Purification and application of α-galactosidase from germinating coffee beans (Coffea arabica)

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Abstract

The objective of this study was to prepare and purify α-galactosidase (α-Gal) from germinating coffee beans. The molecular weight of purified α-Gal from germinating coffee beans was determined to be 38.8 kDa by SDS-PAGE. Then the purified α-Gal was applied to synthesize galactosyl β-cyclodextrin using melibiose as donor and β-cyclodextrin as acceptor. Galactosyl β-cyclodextrin was isolated by preparative high-performance liquid chromatography (HPLC) and its structure was analyzed and elucidated by HPLC, fourier transform infrared spectrometer, electro spraying ionization-mass spectrometry, and 13C nuclear magnetic resonance. Our results strongly demonstrate that the synthesized product is a mono-6-O-α-d-galactopyranosyl-β-cyclodextrin and the purified α-galactosidase could transfer one galactosyl residue directly to the β-cyclodextrin ring and synthesize the mono-6-O-α-d-galactosyl β-cyclodextrin.

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Abbreviations

α-Gal:

α-d-Galactosidase

CD:

Cyclodextrin

SDS-PAGE:

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis

HPLC:

High-performance liquid chromatography

FTIR:

Fourier transform infrared spectrometer

ESIMS:

Electron spray ion mass spectroscopy

NMR:

Nuclear magnetic resonance

DEAE:

Diethyl aminoehtyl

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Acknowledgments

This work was supported by the International Cooperation Project of China and Netherlands (2007DFA31120), the National Key Technology Research and Development Program of the 11th 5-year Plan of China (2006BAD05A01, 2006BAD05A07), and Jiangsu Provincial Natural Science Foundation (BK2008003).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, China

    Wangyang Shen, Hanqing Chen, Zhengyu Jin, Xueming Xu, Jianwei Zhao & Zhengjun Xie

  2. School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, China

    Wangyang Shen, Hanqing Chen, Zhengyu Jin, Xueming Xu, Jianwei Zhao & Zhengjun Xie

  3. Department of Plant Science, North Dakota State University, Fargo, ND, 58105, USA

    Yin Li

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  1. Wangyang Shen
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  2. Yin Li
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  4. Zhengyu Jin
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  5. Xueming Xu
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Correspondence to Zhengyu Jin.

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Shen, W., Li, Y., Chen, H. et al. Purification and application of α-galactosidase from germinating coffee beans (Coffea arabica). Eur Food Res Technol 228, 969–974 (2009). https://doi.org/10.1007/s00217-009-1013-4

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  • DOI: https://doi.org/10.1007/s00217-009-1013-4

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