Diacetyl and acetoin are key aroma components of fermented milk but are produced in low concentrations by starter cultures. In this study, we expressed NADH oxidase, acetolactate synthase, and inactivated acetolactate decarboxylase in Lacticaseibacillus casei TCS to generate recombinant L. casei strains, and investigated the effects of the genes encoding these enzymes on diacetyl and acetoin production during milk fermentation. In the single-gene recombinant strains tested, diacetyl concentrations were highest in milk fermented by L. casei TCSI-nox (nox gene overexpressed, 3.68 mg/kg), whereas acetoin concentrations were highest in milk fermented by L. casei TCS-∆alsD (alsD gene deleted, 32.94 mg/kg). Moreover, diacetyl and acetoin concentrations were higher in the inducible strains than in the corresponding constitutive strains (e.g., TCSInox vs. TCSC-nox, and TCSI-∆alsD-nox vs. TCSC-∆alsD-nox). This phenomenon was also reflected in the protein expression levels and enzyme activities. In the double-gene recombinant strains tested, the highest concentrations of diacetyl and acetoin were produced by L. casei TCSI-∆alsD-nox (nox overexpressed and alsD deleted, 4.66 mg/kg, 69.62 mg/kg, respectively). The triple-gene recombinant L. casei TCS-∆alsD-nox-alsS produced the highest concentrations of diacetyl and acetoin, which were 2.38 and 11.19 times, respectively, the concentrations produced by the original strain. These results show that the nox, alsS, and alsD genes make key contributions to the biosynthesis of diacetyl and acetoin by L. casei. The modification of multiple genes had a synergistic effect, leading to greatly in-creased synthesis of diacetyl and acetoin by L. casei during its fermentation of milk.

Genome-scale metabolic models and flux balance analysis (FBA) have been extensively used for modeling and designing bacterial fermentation. However, FBA-based metabolic models that accurately simulate the dynamics of coculture are still rare, especially for lactic acid bacteria used in yogurt fermentation. To investigate metabolic interactions in yogurt starter culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, this study built a dynamic metagenome-scale metabolic model which integrated constrained proteome allocation. The accuracy of the model was evaluated by comparing predicted bacterial growth, consumption of lactose and production of lactic acid with reference experimental data. The model was then used to predict the impact of different initial bacterial inoculation ratios on acidification. The dynamic simulation demonstrated the mutual dependence of S. thermophilus and L. d. bulgaricus during the yogurt fermentation process. As the first dynamic metabolic model of the yogurt bacterial community, it provided a foundation for the computer-aided process design and control of the production of fermented dairy products.

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Summary This study aimed to investigate changes in the quality of heat‐processed beef flavour (HPBF) during 168‐day storage period at 4, 20 and 50 °C through evaluating 10 physicochemical indexes and sensory profiles. The sensory score of HPBF reduced dramatically at 50 °C compared with those at other temperatures. The correlation analysis indicated that among the 10 physicochemical indexes, oxidation–reduction potential (ORP), which is positively correlated to lipid oxidation, possessed the strongest association with the total sensory score of HPBF. The kinetic analysis showed that the increasing rate of ORP during the first 50‐day period at 50 °C was more than 10 times larger than those at 4 and 20 °C. These results suggested that lipid oxidation played a major role in affecting the quality of HPBF, especially during the early storage period. Reducing lipid oxidation via for instance maintaining a relatively low storage temperature should be given better attention for obtaining high‐quality HPBF.

In this study, the effect of heteroaggregation (HA) on the physicochemical stability and the formation of volatile substances of DHA emulsions was investigated. HA-DHA emulsions were produced by combination of lactoferrin (LF)-DHA and soy protein isolate (SPI)-DHA emulsions at pH 6.0. Zeta-potentials, droplet sizes, stability, and microstructures were measured as a function of different ratios of LF-DHA to SPI-DHA droplets. DHA oxidation of single and HA emulsions was determined through measurements of lipid hydroperoxides, thiobarbituric acid reactive substances, and the formation of volatile substances. LF-DHA to SPI-DHA droplets ratios of 5:5, 4:6, and 3:7 formed stable emulsions. The lowest zeta-potential, biggest droplet size, and optimum physical stability of heteroaggregated emulsion occurred at a 5:5 of LF-DHA to SPI-DHA droplet ratio. Microstructure behavior indicated that the HA emulsions (LF-DHA droplets/SPI-DHA droplets = 5:5) formed specific three-dimensional uniform networks. The formation of thiobarbituric acid reactive substances, lipid hydroperoxides, and volatile compounds including hexanal and (E,E)-2,4-heptadienal decreased in HA compared to single emulsions. The results indicated that the physicochemical stability of DHA emulsions was enhanced and that the formation of volatile substances was inhibited by HA. It thus demonstrated the utilization of HA to improve the stability of bioactive compounds in emulsions.