Isolation of Bowman-Birk-Inhibitor from soybean extracts using novel peptide probes and high gradient magnetic separation

Abstract

Publication informationFood Chemistry, 134 (4): 1831-1838Publisher Elsevier Item record/more information http://hdl.handle.net/10197/8385 Publisher's statementThis is the author's version of a work that was accepted for publication in Food Chemistry. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Food Chemistry (VOL 134, ISSUE 4, (2012) AbstractSoybean proteins offer exceptional promise in the area of cancer prevention and treatment.Specifically, soybean Bowman-Birk inhibitor (BBI) has the ability to suppress carcinogenesis in vivo, which has been attributed to BBI's serine protease activity, made possible by the presence of two distinct binding sites for trypsin and chymotrypsin. The lack of molecular tools for the isolation and identification of this protein has made it difficult to work with, limiting progress as a significant candidate in the treatment of cancer. This study has successfully identified a set of novel synthetic peptides targeting the Bowman-Birk inhibitor, using a phage display screening approach, and has demonstrated the ability to isolate BBI from crude mixtures. These peptide probes have been covalently immobilized on superparamagnetic (SPM) microbeads to allow for high gradient magnetic purification from crude soy whey mixtures in a single step. Our ultimate goal is for the described probe to be utilised to facilitate the isolation of this therapeutically relevant protein for low cost, scalable analysis and production of BBI.

Plant Protease Inhibitors in Therapeutics-Focus on Cancer Therapy

“…Whereas KTIs are 8000–22,000 Da proteins with two disulphide linkages and a single reactive site of trypsin (Laskowski and Qasim, 2000). Among all, BBI is identified as a suitable protein to work with, in terms of handling, resistance to temperature extremities and acidification (Fields et al, 2012). Naturally occurring plant based BBI is considered as a therapeutically significant candidate in the treatment of multiple diseases, especially in the field of cancer prevention (Fields et al, 2012).…”

“…Among all, BBI is identified as a suitable protein to work with, in terms of handling, resistance to temperature extremities and acidification (Fields et al, 2012). Naturally occurring plant based BBI is considered as a therapeutically significant candidate in the treatment of multiple diseases, especially in the field of cancer prevention (Fields et al, 2012). Perhaps its greatest potential in therapeutic property lies in its ability to suppress carcinogenesis irreversibly even during early stage (Wattenberg, 1992).…”

“…Perhaps its greatest potential in therapeutic property lies in its ability to suppress carcinogenesis irreversibly even during early stage (Wattenberg, 1992). Encouragingly, the amount of BBI that reaches the internal organs after oral ingestion reaches the range that prevents malignant transformation in vitro (Yavelow et al, 1985; Fields et al, 2012). Several PPIs are under further evaluation in human clinical trials.…”

Selective Isolation of Trypsin Inhibitor and Lectin from Soybean Whey by Chitosan/Tripolyphosphate/Genipin Co-Crosslinked Beads

“…Several purification approaches focused on trypsin inhibitors and lectins have been proposed; they include electrophoresis [9], affinity chromatography [10,11,12,13], ion exchange chromatography [14], liquid-liquid extraction using reversed micelles [15], aqueous two-phase systems [16], integration of affinity precipitation with aqueous two-phase affinity extraction [17] and superparamagnetic microbeads method [18,19]. An economical, quick and effective technique to purify trypsin inhibitors and lectins would benefit not only biological research, but also industrial applications.…”

Flow enhanced non-linear magnetophoretic separation of beads based on magnetic susceptibility

“…[1][2][3][4][5] For example, antibody and glass functionalized SPM beads are now widely used for the isolation of specific cell types and nucleic acids, respectively. 6,7 High-gradient magnetophoresis (HGM) is 25 typically used to collect the SPM beads from complex aqueous solutions, i.e., a high magnetic field and field gradients generated by a magnet are applied to the suspension to remove the SPM beads.…”