Two-Step Isolation, Purification, and Characterization of Lectin from Zihua Snap Bean (Phaseolus vulgaris) Seeds

Abstract

A two-step method based on an aqueous two-phase system and Sephadex G-75 was used to separate and purify lectin from the seeds of the Zihua snap bean. The preliminary properties and bioactivity of the Zihua snap bean lectin were characterized by different instrumental methods, such as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE), liquid chromatography-nano electrospray ionization mass spectrometry (Nano LC-ESI-MS/MS), and Fourier transform infrared spectroscopy (FTIR). The hemagglutinating activity of the Zihua snap bean lectin could not be inhibited by glucose, N-acetyl-d-glucosamine, d-galactose, N-acetyl-d-galactosamine, fructose, sucrose, d-maltose, d-trehalose, and lactose. It was found that the hemagglutinating activity of the lectin showed strong dependence on Mn2+ and Ca2+. The thermal and pH stability of the Zihua snap bean lectin was studied by FTIR and fluorescence spectroscopy. Relatively good stability was observed when the temperature was not higher than 70 °C, as well as in the pH range of 2.0 to 10.0. Digestive stability in vitro was investigated. The untreated lectin was relatively stable to pepsin and trypsin activity, but heat treatment could significantly reduce the digestive stability in vitro. Moreover, the lectin showed an inhibitory effect on the tested bacteria (Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Bacillus subtilis (B. subtilis)), and it also showed a certain inhibitory effect on the growth of Phytophthora infestans (P. infestans) at higher concentrations.

Efficacy of Live Attenuated Vaccine and Commercially Available Lectin against Avian Pathogenic E. coli Infection in Broiler Chickens

“…The high prevalence of multidrug-resistant APEC poses a zoonotic risk in developing countries [ 8 , 9 , 10 ]. Therefore, alternatives such as vaccines and lectins are being developed to combat E. coli infection in poultry [ 11 , 12 ]. In Egypt, two commercially available live attenuated E. coli vaccines are currently used, however, their efficacy against prevalent homologous and heterologous Egyptian APEC serogroups needs further evaluation.…”

“…Recently, research has been directed toward the therapeutic and preventive applications of lectins due to their antimicrobial effects as an alternative against antibiotic-resistant microorganisms [ 17 ]. A new lectin from fruiting bodies of the mushroom showed antibacterial activity against E. coli [ 12 ]. Additionally, lectins had an immunomodulatory activity by activating macrophages and tumor necrosis factor (TNF), inducing IL-2 and IFN-γ genes expression, thus upregulating the T-helper-1 cell population [ 18 ].…”

Reutilization of Food Waste: One-Step Extration, Purification and Characterization of Ovalbumin from Salted Egg White by Aqueous Two-Phase Flotation

“…Furthermore, Nano LC-ESI-MS/MS is a reliable and sensitive method for identifying gel-separated protein tapes. Moreover, a single protein tape can be identified in mixed proteins by this method [37]. Table 5 and Table 6 showed the results of the OVA purified from the phase of ATPF determined by Nano LC-ESI-MS/MS.…”

Study on the Preparation and Conjugation Mechanism of the Phosvitin-Gallic Acid Complex with an Antioxidant and Emulsifying Capability

“…The Pv–GA complex was characterised by infrared spectroscopy and its derivative methods. The spectral differences between different substances were effectively distinguished by FTIR [36,37]. The infrared spectra of Pv and the Pv–GA complex are shown in Figure 3.…”