The effect of chickpea flour and its addition levels on quality and in vitro starch digestibility of corn-rice-based gluten-free pasta.

Development of gluten-free (GF) pasta with improved nutritional attributes is one of the main trends in the gluten-free pasta industry. Considerable interest lays in introducing legume-based ingredients into traditional corn/rice GF formulations. This work aims to fortify multi-cereal (corn-rice) GF pasta with chickpea to investigate how different chickpea addition levels affect its quality and in vitro starch digestibility. Chickpea significantly increased pasta protein and dietary fibre contents to a level that supports the "source" or "high" fibre/protein content claims. Chickpea addition induced darkening, softening, adhesiveness decrease and solid loss reduction compared to the control. In addition, chickpea substitution significantly modified the in vitro starch digestion, which showed increasing resistant starch and decreasing slowly digestible starch contents suggesting potential mitigation of postprandial glucose response in vivo. Reformulating GF pasta with chickpea flour should, therefore, be considered as an effective tool to improve the corn-rice-based GF products' nutritional profile.

Antigenic activity and epitope analysis of ß-conglycinin hydrolyzed by pepsin.

BACKGROUND: Soybean is among the 'big eight' allergenic foods, and ß-conglycinin, the main antigenic protein of soybean, has high levels of antigenic activity. Why the antigenic activity of soybean ß-conglycinin is not eliminated by enzymatic hydrolysis is not clear. In this study, changes in the molecular composition and antigenicity of ß-conglycinin hydrolyzed by pepsin were analyzed and it was determined whether complete sequential epitopes exist in the resulting hydrolysates. The nature and antigenic activity of protein subunits obtained after ß-conglycinin hydrolysis were also assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and competitive enzyme-linked immunosorbent assay, respectively. RESULTS: The residual antigenic activity of ß-conglycinin was 52%, α'- and α-subunits completely disappeared, the 49 kDa fraction partially disappeared, and peptides measuring 27 and 23 kDa were newly formed after 60 min of enzymatic hydrolysis. Prolonged enzymatic hydrolysis did not result in remarkable changes in these peptides; thus, the peptides show some resistance to enzymatic hydrolysis. The amino acid sequences of the peptide chains were analyzed by matrix-assisted laser desorption / ionization-time of flight mass spectrometry and aligned with the related sequences in the corresponding protein and antigen databases. Ten complete sequential epitopes were identified in the residual 49 kDa fraction, of these epitopes, two were from α-subunits and eight were from ß-subunits. CONCLUSION: The presence of complete sequential epitopes in hydrolysates obtained from the enzymatic hydrolysis of soybean is an important reason for the incomplete disappearance of the antigenic activity of ß-conglycinin. © 2020 Society of Chemical Industry.

Effects of ultraviolet-C light-emitting diodes at 275 nm on inactivation of Alicyclobacillusacidoterrestris vegetative cells and its spores as well as the quality attributes of orange juice.

Alicyclobacillus acidoterrestris is a thermoacidophilic, spore-forming bacillus. A. acidoterrestris and its spores can survive in pasteurized juices and cause microbial spoilage. In this work, the effects of ultraviolet-C light-emitting diodes at 275 nm on the inactivation of A. acidoterrestris vegetative cells and its spores in commercial pasteurized orange juice were studied. Meanwhile, the effects of ultraviolet-C light-emitting diodes on the quality attributes of the orange juice were also investigated. The quantities of A. acidoterrestris vegetative cells and its spores inoculated in orange juice were reduced by 6.04 and 2.49 log10 CFU/mL after ultraviolet-C light-emitting diode treatment at 220 mJ/cm2, respectively. The Weibull and Weibull plus tail models were satisfactorily fitted to estimate the reductions of A. acidoterrestris vegetative cells and its spores in orange juice, respectively. Physicochemical properties (pH, titratable acidity, total soluble solids, and clarity) of orange juice did not change significantly after exposure to ultraviolet-C light-emitting diodes. However, the total phenolic content of orange juice decreased with increasing fluence. In addition, ultraviolet-C light-emitting diode treatment at a higher fluence led to a noticeable color difference. These results indicate that ultraviolet-C light-emitting diode treatment has a potential application in the juice processing industry.

Recombinase polymerase amplification with polymer flocculation sedimentation for rapid detection of Staphylococcus aureus in food samples.

Currently, rapid, sensitive, and convenient visual detection methods for Staphylococcus aureus (S. aureus) are scarce. In this study, a novel detection method based on recombinase polymerase amplification (RPA) and polymer flocculation sedimentation (PFS) was developed. Twelve effective primer combinations derived from four forward primers F1, F2, F3, F4, and three reverse primers R1, R2, R3 targeting the nuc gene of S. aureus were designed and screened by a polymerase chain reaction and RPA methods. RPA reaction conditions, including temperature, time, and volume as well as PEG8000 and NaCl concentrations range, were optimized. Moreover, the specificity and sensitivity of the RPA-PFS assay were further analyzed. Finally, the potential use of the RPA-PFS assay was evaluated using artificially S. aureus contaminated food samples, including pork, beef, shrimp, fish, cheese, cabbage, leftover rice, egg, milk, and orange juice. Results showed that the SA5 (F2/R2) combination was the optimal primer candidate. The optimal temperature range, the shortest time and the minimal volume of RPA reaction were 40-42 °C, 10 min and 10 µL, respectively and the optimal PEG8000/NaCl concentrations were 0.2 g/mL and 2.5 M, respectively, for the adsorption between magnetic beads and RPA products. The RPA-PFS method could detect as little as 13 fg genomic DNA of S. aureus and was also specific for five target S. aureus as well as twenty-seven non-target foodborne bacteria. The limit of detection of RPA-PFS for S. aureus in artificially contaminated food samples was 38 CFU/mL (g). Besides, RPA-PFS has directly been judged by the naked eye and has totally taken less than 20 min. In short, the assay RPA-PFS developed in this study is a rapid, sensitive, and specific visual detection method for S. aureus.

Rapid analysis of Escherichia coli O157:H7 using isothermal recombinase polymerase amplification combined with triple-labeled nucleotide probes.

Rapid analytical methods are urgently needed to evaluate Escherichia coli (E. coli) O157:H7 in food. In this work, a novel recombinase polymerase amplification (RPA)-based lateral flow dipstick (LFD) method was developed to detect E. coli. Briefly, suitable primers and probes were designed and screened. Then, RPA reaction parameters, including volume, time, and temperature, were optimized. The specificity and sensitivity of RPA-LFD were analyzed, and a contaminated milk sample was used to test the detection performance of the proposed method. The optimal RPA reaction conditions included a minimum volume of 10 µL, incubation time of 10 min, temperature range of 39-42 °C, the primer pair EOF4/EOR3, and the probe EOProb. RPA-LFD was highly sensitive, it could detect as little as 1 fg of the genomic DNA of E. coli O157:H7, and 19 nontarget DNA of foodborne bacteria did not yield amplification products. Finally, the limit of detection of RPA-LFD for E. coli O157:H7 in artificially contaminated raw milk was 4.4 CFU/mL. In summary, the RPA-LFD assay developed in this study is an effective tool for the rapid investigation of E. coli O157:H7 contamination in raw milk samples.

Sensitive and rapid visual detection of Salmonella Typhimurium in milk based on recombinase polymerase amplification with lateral flow dipsticks.

Salmonella Typhimurium (S. Typhimurium) can cause serious foodborne diseases. In this study, an assay combining recombinase polymerase amplification (RPA) with lateral flow dipsticks (LFD) was developed to detect S. Typhimurium in milk. The RPA forward primers STF1, STF2, STF3, the reverse primer STR labeled with digoxin, and the probe STProb labeled with FAM were designed and screened to produce RPA products for LFD detection. The RPA reaction volume, temperature, and time were then optimized, and the sensitivity and specificity of the developed method were analyzed. Finally, the RPA-LFD method was evaluated using milk artificially contaminated with S. Typhimurium. Results indicated that the primer pair STF1/STR is the optimal combination for detecting the bacterium. The minimum volume, shortest time, and optimal temperature of the RPA reaction were 10 µL, 10 min, and 40-42 °C, respectively. The limit of detection of RPA-LFD for detecting the genomic DNA of S. Typhimurium was 1 fg, which is 5 and 10 times lower than the corresponding limits of RPA-agarose gel electrophoresis (AGE) and PCR-AGE, respectively. Testing with 29 other foodborne bacteria as controls revealed that RPA-LFD was highly specific for S. Typhimurium. RPA-LFD can detect S. Typhimurium at concentrations as low as 1.95 CFU/mL in artificially inoculated milk samples and is thus 10 times more sensitive than PCR. Hence, the RPA-LFD assay established in this study could be a potential point-of-care/need test for S. Typhimurium, especially in areas with limited resources.

Development of duplex PCR-ELISA for simultaneous detection of Salmonella spp. and Escherichia coli O157: H7 in food.

In the current study, a duplex PCR-ELISA method was developed targeting the specific genes, invA of Salmonella spp. and rfbE of Escherichia coli O157: H7, to detect one or both bacteria in food. In brief, PCR product amplified by PCR primer labeled with digoxin at the 5'-end and a probe labeled with biotin at the 3'-end can form dimer by nucleic acid hybridization which can be captured by binding of biotin to streptomycin coated in ELISA plate before using enzyme-labeled anti-digoxin antibody and substrate to develop color. Also, evaluation of the duplex PCR-ELISA method was conducted in different food samples including milk, juice, cabbage, shrimp, chicken, pork and beef. Results indicated that the duplex PCR-ELISA developed here was specific when using 25 non-target bacteria strains as controls and was sensitive with a limit of detection (LOD) of 1 CFU/mL, 1, 000 times higher than that of the duplex PCR method and was repeatable regardless of inter- and intra-batch variations. The duplex PCR-ELISA method established in the present study has proven to be highly specific, sensitive and repeatable. It has the potential to be applied in such fields as clinical diagnosis of food-borne diseases, food hygiene monitoring and pathogen detection in food.

Reaction kinetics in food extrusion: methods and results.

Extrusion cooking is a highly efficient food processing technology. During the extrusion process, there are many desirable and undesirable reactions which will determine final product quality. While being heated and sheared simultaneously, food raw materials experience a non-isothermal process and their residence time in the extruder is distributed. All these factors contribute to the difficulties in determining the kinetic parameters for those reactions. Therefore, this paper attempts to review the reaction kinetics in food extrusion. First of all, the kinetic models for the reactions are outlined. After elucidating how to determine reaction time in an extruder, the methodological approaches for determining the reaction order, rate constant, and activation energy of a reaction under isothermal or non-isothermal conditions with or without residence time distribution (RTD) are presented. Then, different models relating the rate constant to its various impact factors, with especially focusing on shear stress, are reviewed. Subsequently, how shear stress is estimated in an extruder, is illustrated. In the last part of this paper, the reported data of rate constant, reaction order, and activation energy for the reactions occurring during food extrusion are summarized, with detailed impacts of temperature, moisture content, shear stress, and determination method on these kinetic parameters. Finally, future research needs are suggested.

Mechanochemistry in thermomechanical processing of foods: kinetic aspects.

In some food processing operations, raw materials sustain shearing and heating simultaneously. These thermomechanical treatments will induce chemical, physicochemical, and biochemical changes, such as starch degradation and gelatinization, protein denaturalization, loss of nutritious components, and inactivation of enzymes and their inhibitors. In this article, only kinetic aspects of these changes are reviewed. The Basedow and Zhurkov models are commonly used to describe shear effects on the rate constants of mechanochemical reactions. After a brief description of these 2 models, their applications in food are reviewed deeply, with emphasis on the activation energy reduction in the Zhurkov model and the shear activation energy in the Basedow model. Since the changes occurring in food systems often involve various interactions at the atomic, molecular, and supramolecular levels, they were described only phenomenonlogically by the mechanochemical kinetic models. The limitation, opportunity, and extension of the mechanochemical approach to modeling food kinetics are discussed.