Isolation of soybean protein P34 from oil bodies using hydrophobic interaction chromatography

Abstract

Background: Soybeans play a prominent role in allergologic research due to the high incidence of allergic reactions. For detailed studies on specific proteins it is necessary to have access to a large amount of pure substance.

Expression of the soybean allergenic protein P34 in Escherichia coli and its indirect ELISA detection method

“…In previous studies, P34 was extracted from soybean oil bodies using hydrophobic interaction chromatography (Sewekow et al 2008). A subsequent purification, however, is in any case necessary.…”

“…IgE binding assays using immunoglobulin from soybean-sensitive individuals indicated that 65 % of the total allergenic response was induced by P34 (Helm et al by disulfide linkages in the 7S globulin fraction and may play a role in protein folding (Helm et al 2000). As a thiol protease, P34 is a member of the papain superfamily (Sewekow et al 2008) and has a tendency to bind to lipid and oil, which is likely significant in its role as an allergen (Mills et al 2004). P34 exhibits post-translational production derived from a 46-to 47-kDa precursor protein by the partial removal of 122 Nterminal amino acid residues, and its tertiary conformation was similar to the papain family .…”

Review Soybeans as bioreactors for biopharmaceuticals and industrial proteins

“…For the latter, the 15S fraction corresponds to 1% of the globulins, has a high molecular mass (700 kDa) and has an isoelectric point of 4.5. Furthermore, the known proteins in soybean seeds with high allergenic potential are mainly composed of a 34-kDa maturing seed protein (also known as P34, Gly m Bd 30 K or Gly m 1) (Sewekow et al, 2008), the 23-kDa protein Gly m Bd 28 K (Xiang et al, 2004), β-conglycinin (Krishnan et al, 2009), the glycinin Gly m 6 (Holzhauser et al, 2009), lectin andlypoxygenase (L'Hocine andBoye, 2007), and a Kunitztype protease inhibitor (Roychaudhuri et al, 2004). There are also some allergenic proteins in seed shell, such as Gly m 1.0101, Gly m 1.0102, Gly m 2, Gly m 3, and SAM 22 (Gly m 4).…”

Análise proteômica de variedades convencionais e geneticamente modificadas de soja (Glycine max) visando proteínas bioativas

“…Trata-se de uma protease que pertence à superfamília papaína. Nas células vegetais, apresenta-se associada a frações lipídicas e localiza-se no interior de vacúolos de proteínas de armazenamento (SEWEKOW et al, 2008;KALINSKI et al, 1992). Muitas proteases tióis estão associadas à germinação, ou seja, na degradação protéica e mobilização de proteínas para o desenvolvimento da planta.…”

“…A P34 é reportada como uma dos principais alérgenos da soja em seres humanos sensíveis e corresponde a cerca de 2-3% das proteínas da soja (HERMAN et al, 2003, SEWEKOW et al, 2008. Apresenta peso molecular de 28,643 Da e na forma glicosilada, a massa é ligeiramente maior, apresentando uma banda de 32kDa em géis SDS-PAGE (SEWEKOW et al, 2008).…”