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Journal of Biochemistry Advance Access originally published online on March 15, 2008
Journal of Biochemistry 2008 143(6):833-839; doi:10.1093/jb/mvn038
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© 2008 The Japanese Biochemical Society

One-Step Purification of Lectins from Banana Pulp Using Sugar-Immobilized Gold Nano-Particles

Sachiko Nakamura-Tsuruta1, Yuko Kishimoto3, Tomoaki Nishimura3 and Yasuo Suda1,2,3,*

1Venture Business Laboratory; 2Department of Nanostructure and Advanced Materials, Kagoshima University, 1-21-40, Kohrimoto, Kagoshima 890-0065; and 3SUDx-Biotec Corp., KIBC #461, 5-5-2, Minatojima-minami, Chuo-ku, Kobe 650-0047, Japan

*To whom correspondence should be addressed. Tel.: +81-99-285-8369, Fax: +81-99-285-8369, E-mail: ysuda{at}eng.kagoshima-u.ac.jp

Received December 24, 2007; Accepted February 29, 2008


  Abstract

To obtain lectins without tedious purification steps, we developed a convenient method for a one-step purification of lectins using sugar-immobilized gold nano-particles (SGNPs). Proteins in crude extracts from plant materials were precipitated with 60% ammonium sulphate, and the precipitate was re-dissolved in a small volume of phosphate buffer. The resultant solution was then mixed with appropriate SGNPs under an optimized condition. After incubating overnight at 4°C, lectins in the mixture formed aggregate with SGNPs, which was visually detected and easily sedimented by centrifugation. The aggregate was dissolved by adding inhibitory sugars, which were identical to the non-reducing sugar moieties on the SGNPs. According to SDS–PAGE and MS of thus obtained proteins, it was found that SGNPs isolated lectins with a high purity. For example, a protein isolated from banana using Glc{alpha}-GNP ({alpha}-glucose-immobilized gold nano-particle) was identified as banana lectin by trypsin-digested peptide-MS finger printing method.

Key Words: gold nano particle, lectin, peptide MS fingerprinting, purification, sugar chain

Abbreviations: CHCA, {alpha}-cyano-4-hydroxycinnamic acid; Glc{alpha}-GNP, alpha-glucose-immobilized gold nano-particle; GlcNAc{alpha}-GNP, alpha-N-acetyl-glucosamine-immobilized gold nano-particle; Man{alpha}-GNP, alpha-mannose-immobilized gold nano-particle; SA, 3,5-dimethyl-4-hydroxycinnamic acid; SGNPs, sugar-immobilized gold nano-particles


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