Logistic modeling to predict the minimum inhibitory concentration (MIC) of olive leaf extract (OLE) against Listeria monocytogenes.

Olive leaf extract (OLE) has been increasingly recognized as a natural and effective antimicrobial against a host of foodborne pathogens. This study attempts to predict the minimum inhibitory concentration (MIC) of OLE against Listeria monocytogenes F2365 by utilizing the asymptotic deceleration point (PDA) in a logistic model (LM), namely MIC-PDA. The experimental data obtained from the inhibitory rate (IR) versus OLE concentration against L. monocytogenes were sufficiently fitted (R2 = 0.88957). Five significant critical points were derived by taking the multi-order derivatives of the LM function: the inflection point (PI), the maximum acceleration point (PAM), the maximum deceleration point (PDM), the absolute acceleration point (PAA), and the asymptotic deceleration point (PDA). The PDA ([OLE] = 37.055 mg/mL) was employed to approximate the MIC-PDA. This MIC value was decreased by over 42% compared to the experimental MIC of 64.0 mg/mL, obtained using the conventional 2-fold dilution method (i.e., MIC-2fold). The accuracy of MIC-PDA was evaluated by an in vitro L. monocytogenes growth inhibition assay. Finally, the logistic modeling method was independently validated using our previously published inhibition data of OLE against the growths of Escherichia coli O157:H7 and Salmonella enteritidis. The MIC-PDA (for [OLE]) values were estimated to be 41.083 and 35.313 mg/mL, respectively, compared to the experimental value of 62.5 mg/mL. Taken together, MIC-PDA, as estimated from the logistic modeling, holds the potential to shorten the time and reduce cost when OLE is used as an antimicrobial in the food industry.

Chemical composition as an indicator for evaluating heated whey protein isolate (WPI) and sugar beet pectin (SBP) systems to stabilize O/W emulsions.

We investigated changes in the chemical composition of WPI as a result of heating (60 °C, 72 h) with SBP in solution (pH 6.75). The concentration of WPI was kept at a constant (3%), whereas the level of SBP was varied at 1, 1.5, and 3%. The reaction products were examined using the Ellman's reagent, ninhydrin assay, and gel electrophoresis. The results demonstrated that the losses of the free sulfhydryl (-SH) and primary amine (-NH2) contents in WPI were less severe compared to those occurring in the dry-state at similar conditions (mass ratio, temperature, and reaction duration). The mixtures were used as emulsifiers in an O/W emulsion system at pH 3.20 and 6.75 and showed an improved ability to stabilize the average size of the droplets than WPI alone at acidic pH. The mixtures at higher levels of SBP, ≥ 1.5%, however, adversely affected the emulsion stability at neutral pH.

Structural and thermal stability of ß-lactoglobulin as a result of interacting with sugar beet pectin.

Changes in the structural and thermal stability of ß-lactoglobulin (ß-LG) induced by interacting with sugar beet pectin (SBP) have been studied by circular dichroism (CD), Fourier transform infrared, and steady-state as well as time-resolved fluorescence spectroscopic techniques. It has been demonstrated that SBP not only is capable of binding to native ß-LG but also causes a significant loss in antiparallel ß-sheet, ∼10%, accompanied by an increase in either random coil or turn structures. In addition, the interaction also disrupted the environments of all aromatic residues including Trp, Phe, and Tyr of ß-LG as evidenced by near-UV CD and fluorescence. When preheated ß-LG was combined with SBP, the secondary structure of ß-LG was partially recovered, ∼4% gain in antiparallel ß-sheet, and Trp19 fluorescence was recovered slightly. Although forming complexes with SBP did not significantly impact the thermal stability of individual secondary structural elements of ß-LG, the environment of Trp19 was protected considerably.

Investigation of molecular interactions between ß-lactoglobulin and sugar beet pectin by multi-detection HPSEC.

Molecular interactions between ß-lactoglobulin (ß-LG) and sugar beet pectin (SBP) were studied using online multi-detection high performance size exclusion chromatography (HPSEC) at neutral pH and 50mM ionic strength. The hydrodynamic properties of various interacting polymer fractions were characterized in detail and compared with those of ß-LG and SBP. Results showed that ∼6.5% (w/w) of native dimeric ß-LG molecules formed complexes with over 35% SBP molecules of varying sizes, 800, 110 and 75 kDa. Although the ß-LG molecules bind to SBP molecules of all sizes and shapes, they tend to favor the intermediate (110 kDa) and small sized (75 kDa) SBP molecules. All resulting complexes possess altered shapes and hydrodynamic properties when compared to unbound SBP and ß-LG. About half of the interacting ß-LG (∼3.5%) molecules were thought to bind to a small amount of non-covalently bound feruloyl groups, possibly present in SBP. When pre-heat treated ß-LG and SBP were combined, more than 16% of ß-LG formed complexes with at least 45% of SBP molecules of varying sizes, Mw∼750-800, 110, and 55-80 kDa. The complexes formed between ß-LG aggregates and/or oligomers and the large SBP molecules (750-800 kDa) adopt the shape of ß-LG aggregates, random coil. Both groups of complexes formed between ß-LG intermediate (110 kDa) and small sized (55-80 kDa) SBP take on the shape of rigid rod. It was speculated that half of the interacting heat-treated ß-LG molecules (∼8%) are complexed with non-covalently bound feruloyl groups in SBP.

Physico-chemical characterization of protein-associated polysaccharides extracted from sugar beet pulp.

We have solubilized and separated polysaccharides from sugar beet pulp (SBP) into three fractions with steam assisted flash extraction (SAFE). For pectin, recovery ranged from 8 to 14%, degree of methy-esterification 66-73%, crude protein 1.3-1.7%, M(w) 262-318 kDa, η(w) 0.22-0.23 dL/g, Rg(z) 36-39 nm and Rh(z) 41-42 nm. For alkaline soluble polysaccharides, (ASP I) recovery ranged from 4.0 to 6.5%, crude protein 1.2-4.8%, weight average molar mass (M(w)) 66-68 kDa, weight average intrinsic viscosity (η(w)) 0.27-0.30 dL/g, z-average radius of gyration (Rg(z)) 25-29 nm and z-average hydrated radius (Rh(z)) 10-11 nm. ASP II recovery ranged from 2.0 to 8.6%, crude protein 1.2-4.8%, M(w) 299-339 kDa, η(w) 0.22-0.33 dL/g, Rg(z) 33-34 nm and Rh(z) 30-34 nm. Recovery of the residue mainly cellulose, ranged from 20.3 to 22.3%. The cellulose in this fraction was converted to carboxymethyl cellulose (CMC). The CMC fraction contained 0.33-0.43 crude protein and had an M(w) ranging from 127 to 263 kDa, η(w) 3.6-8.0 dL/g, Rg(z) 35-45 nm and Rh(z) 27-40 nm.