The number of live lactic acid bacteria (LAB) is an important quality indicator for yogurt, the quantitative testing of LAB has become an important task in the evaluation of product quality and function. By analyzing and comparing the performance of 16S rRNA gene and tuf gene used in absolute quantification, the tuf gene with copy number 1 was selected as the target gene of six LAB. By drawing a standard curve to achieve qualitative and quantitative detection of six strains of LAB, the detection range was found to be 1 × 103–1 × 108 copies/µL. The traditional plate colony count and Flow Cytometry (FCM) were compared with the method of qPCR, which was used in this experiment. Meanwhile, the confocal laser microscope combined with STYO 9 and propidium iodide dyes was used to determine that the content of viable bacteria in the yogurt was more than 90%, which proved that the detection result using qPCR method was closer to the true level of LAB in yogurt. Compared with the existing methods, the method in this study allowed the qualitative and quantitative detection of the six kinds of LAB in yogurt, and the distribution of live and dead bacteria in yogurt could be calculated.
Goose liver oil (GLO) microcapsules were prepared by konjac glucomannan (KGM) and soybean protein isolate (SPI) for the first time as wall materials. The GLO could be effectively encapsulated, with an encapsulation efficiency of 83.37%, when the ratio of KGM to SPI was 2.9:1, the concentration of the KGM-SPI composite gel layer was 6.28% and the ratio of the GLO to KGM-SPI composite gel layer was 1:6. Fourier transform infrared spectroscopy and X-ray diffraction methods showed electrostatic interactions between KGM and SPI molecules and the formation of hydrogen bonds between the GLO and KGM-SPI wall components. The results of scanning electron microscopy showed a smooth spherical surface morphology of the microcapsules with a dense surface and no cracks. The confocal laser scanning microscopy showed that the microcapsules were homogeneous inside and no coalescence occurred. The encapsulated GLO has a significantly higher thermal and oxidative stability compared to free GLO. In the in vitro digestion experiment, 85.2% of the microcapsules could travel through gastric juice, and 75.2% could be released in the intestinal region. These results suggested that microcapsules prepared by KGM-SPI might be used as a carrier for the controlled release of GLO and could microencapsulate various oil-soluble nutrients in food products.
The interaction of polysaccharides–protein with varied origins and structures provides opportunities for tailoring the physicochemical qualities of food protein-based materials. This work examined the feasibility of ultrasound-modified interaction between kidney bean dietary fiber (KSDF) and duck myofibrillar proteins (MP) to improve the physicochemical properties of the gel matrices. Accordingly, gel strength, water holding capacity, solubility, chemical interaction, secondary structure, and network structure of MP were determined. The addition of KSDF combined with the ultrasound treatment contributed to the improved water retention capability, G’ values, and the reduced particle size of protein molecules, corresponding with the formation of dense pore-like structures. The results demonstrated that 1% KSDF and ultrasonication at 400 W significantly enhanced gel strength by up to 109.58% and the solubility increased by 213.42%. The proportion of α-helices of MP gels treated with 1% KSDF and ultrasonication at 400 W was significantly increased. The sonication-mediated KSDF–MP interaction significantly improved hydrophobic interactions of the proteins, thus explaining the denser network structure of the MP gels incorporated KSDF with ultrasound treatments. These results demonstrated the role of ultrasonication treatments in modifying KSDF–protein interaction to improve the gel and structural properties of the MP gels.
The number of viable lactic acid bacteria (LAB) is a key indicator of the quality of fermented milk. Currently, the combination of propidium monoazide (PMA) and qPCR has been applied in the quantification of viable bacteria in various matrices. In this research, the PMA-qPCR method was used to detect the number of viable bacteria of each LAB species in fermented milk. By analyzing pheS gene and 16S rRNA gene sequence similarities in five species of LAB, namely Lactobacillus delbrueckii subsp. bulgaricus, Lactiplantibacillus plantarum, Streptococcus thermophilus, Lactobacillus helveticus, and Lactococcus lactis subsp. lactis, the pheS gene resolved species identities better and was thus selected to design specific primers and probes. The pheS gene was cloned into the pUC19 vector and used to construct a standard curve for absolute quantification. Standard curves for quantification were constructed for each LAB species for serial dilutions between 1011 and 106 CFU/mL, with R2 > 0.99. The number of viable bacteria in the fermented milk detected by PMA-qPCR was significantly lower than that of qPCR (P < 0.05), indicating that PMA inhibited the amplification of DNA from dead cells. This was corroborated by the results from bacterial staining and plate count experiments. The proposed PMA-qPCR method provided rapid qualitative and quantitative determination of the number of viable bacteria for each LAB species in fermented milk within 3 h.
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