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The aim of this research was to develop and characterize gelatin-chitosan (4:1) based films that incorporate nanoemulsions loaded with a range of active compounds; N1: canola oil; N2: αtocopherol/cinnamaldehyde; N3: α-tocopherol/garlic oil; or N4: a-tocopherol/cinnamaldehyde and garlic oil. Nanoemulsions were prepared in a microfluidizer with pressures ranging from 69 to 100 MPa, and 3 processing cycles. Films were produced by the casting method incorporating 5g N1,2,3,4/100 g biopolymers and using glycerol as a plasticizer, and subsequently characterized in terms of their physico-chemical, antimicrobial and antioxidant properties. No differences (p>0.05) were observed for all films in terms of moisture content (18% w/w), and thermal properties. The films' solubility in water and light transmission at 280 nm were considerably reduced as compared to the control, N1 (15% and 60% respectively) because of the nanoemulsion incorporation. The film loaded with N1 showed the greatest (p<0.05) opacity, elongation at break and stiffness reduction, and was the roughest, whilst the lowest tensile strength and ability to swell were attained by films loaded with N3 and N4, respectively. DSC and X-ray analyses suggested compatibility among the biopolymeric-blend, and a good distribution of nanodroplets embedded into the matrix was confirmed by AFM and SEM analyses. Films loaded with nanoencapsulated active compounds (NAC) were very effective against Pseudomonas aeruginosa, and also showed high antioxidant activity. Overall, the present study offers clear evidence that these active-loaded films have the potential to be utilized as packaging material for enhancing food shelf life.

The use of Tetragenococcus halophilus and Zygosaccharomyces rouxii as starter cultures is essential for desirable volatiles production during moromi stage of soy sauce fermentation. In this study, the effect of simultaneous and sequential inoculation of cultures in moromi fermentation models, with respect to viability, physicochemical changes, and volatiles formation (using SPME-GC/MS) was investigated. Interestingly, an antagonism was observed as T. halophilus only proliferated (3 log increase) in the presence of Z. rouxii, while Z. rouxii growth was suppressed by 4 log in concurrence with pH increase to 7.31. Final content of reducing sugars, ethanol, acetic acid, and amino nitrogen did not differ significantly (p<0.05) between co-inoculation and sequential inoculation. However, Z. rouxii promoted alcohols formation and produced a more complex aroma profile under suppression. According to Principal Component Analysis (PCA), the inoculation sequence (co-inoculation and sequential) has impacts on volatile compound profiles during moromi fermentation.

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