News Column

Investigators at Hiroshima University Describe Findings in Peptides and Proteins (Affinity purification of recombinant proteins using a novel...

July 16, 2014



Investigators at Hiroshima University Describe Findings in Peptides and Proteins (Affinity purification of recombinant proteins using a novel silica-binding peptide as a fusion tag)

By a News Reporter-Staff News Editor at Biotech Week -- Research findings on Peptides and Proteins are discussed in a new report. According to news reporting from Hiroshima, Japan, by NewsRx journalists, research stated, "We recently reported that silica is deposited on the coat of Bacillus cereus spores as a layer of nanometer-sized particles (Hirota et al. 2010 J Bacteriol 192: 111-116). Gene disruption analysis revealed that the spore coat protein CotB1 mediates the accumulation of silica (our unpublished results)."

The news correspondents obtained a quote from the research from Hiroshima University, "Here, we report that B. cereus CotB1 (171 amino acids [aa]) and its C-terminal 14-aa region (corresponding to residues 158-171, designated CotB1p) show strong affinity for silica particles, with dissociation constants at pH 8.0 of 2.09 and 1.24 nM, respectively. Using CotB1 and CotB1p as silica-binding tags, we developed a silica-based affinity purification method in which silica particles are used as an adsorbent for CotB1/CotB1p fusion proteins. Small ubiquitin-like modifier (SUMO) technology was employed to release the target proteins from the adsorbed fusion proteins. SUMO-protease-mediated site-specific cleavage at the C-terminus of the fused SUMO sequence released the tagless target proteins into the liquid phase while leaving the tag region still bound to the solid phase. Using the fluorescent protein mCherry as a model, our purification method achieved 85 % recovery, with a purity of 95 % and yields of 0.60 +/- 0.06 and 1.13 +/- 0.13 mg per 10-mL bacterial culture for the CotB1-SUMO-mCherry and CotB1p-SUMO-mCherry fusions, respectively."

According to the news reporters, the research concluded: "CotB1p, a short 14-aa peptide, which demonstrates high affinity for silica, could be a promising fusion tag for both affinity purification and enzyme immobilization on silica supports."

For more information on this research see: Affinity purification of recombinant proteins using a novel silica-binding peptide as a fusion tag. Applied Microbiology and Biotechnology, 2014;98(12):5677-5684. Applied Microbiology and Biotechnology can be contacted at: Springer, 233 Spring St, New York, NY 10013, USA. (Springer - www.springer.com; Applied Microbiology and Biotechnology - www.springerlink.com/content/0175-7598/)

Our news journalists report that additional information may be obtained by contacting M.A.A. Abdelhamid, Hiroshima University, Grad Sch Adv Sci Matter, Dept. of Mol Biotechnol, Hiroshima 7398530, Japan. Additional authors for this research include K. Motomura, T. Ikeda, T. Ishida, R. Hirota and A. Kuroda (see also Peptides and Proteins).

Keywords for this news article include: Asia, Japan, Hiroshima, Proteomics, Amino Acids, Fusion Proteins, Recombinant Proteins, Peptides and Proteins

Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC


For more stories covering the world of technology, please see HispanicBusiness' Tech Channel



Source: Biotech Week


Story Tools






HispanicBusiness.com Facebook Linkedin Twitter RSS Feed Email Alerts & Newsletters