New application of microbial l-glutaminase as a flavor enhancing agent in beef burgers

L‑glutaminase (L‑glutamine amidohydrolase EC3.5.1.2) is the key enzyme in enhancing the taste and aroma of oriental fermented foods by increasing their glutamic acid content and as a result imparting a palatable taste. Beef burgers were prepared using different levels of the partially purified L- glutaminase (2.0 to10.0 U/100 g) prepared from Aspergillus oryzae NRRL 32567. Beef burgers treated with 6.0 U/100g and the others treated with monosodium glutamate (5000 ppm) were chemically, sensory and microbiologically evaluated and compared to untreated control during frozen storage at –18 ºC for 3 months. Treatment with L‑glutaminase (6 U/100g) resulted in an increase of 443% in glutamic acid and a reduction of 63% in glutamine contents resulting in an enhanced preferable taste and odor of the prepared beef burgers. Burgers treated with 6.0 U/100g exhibited the best odor, texture, taste and overall quality scores when compared to the untreated control and samples treated with monosodium glutamate (5000 ppm). During the frozen storage of all samples, an expected slight, but significant (p≤0.05), increase in the total mesophilic bacterial count was evident and such increase was quite acceptable since numbers did not exceed the limit of 5.7x103 cfu/g. Similarly, the total psychrotrophs did not exceed 3.7x102 cfu/g.

SlCCD1A Enhances the Aroma Quality of Tomato Fruits by Promoting the Synthesis of Carotenoid-Derived Volatiles

The loss of volatiles results in the deterioration of flavor in tomatoes. Volatiles are mainly derived from fatty acid, carotenoid, phenylpropane, and branched chain amino acids. In this study, the tomato accession CI1005 with a strong odor and accession TI4001 with a weak odor were analyzed. The volatile contents were measured in tomato fruits using gas chromatography-mass spectrometry. The scores of tomato taste and odor characteristics were evaluated according to hedonistic taste and olfaction. It was found that the content of fatty acid-derived volatiles accounted for more than half of the total volatiles that had grassy and fatty aromas. Phenylpropane-derived volatiles had irritation and floral aromas. Branched-chain amino acid-derived volatiles had a caramel aroma. Carotenoid-derived volatiles had floral, fruity, fatty, and sweet-like aromas, preferred by consumers. A lack of carotenoid-derived volatiles affected the flavor quality of tomato fruits. The accumulation of carotenoid-derived volatiles is regulated by carotenoid cleavage oxygenases (CCDs). A tissue-specific expression analysis of the SlCCD genes revealed that the expression levels of SlCCD1A and SlCCD1B were higher in tomato fruits than in other tissues. The expression levels of SlCCD1A and SlCCD1B were consistent with the trend of the carotenoid-derived volatile contents. The expression of SlCCD1A was higher than that for SlCCD1B. A bioinformatics analysis revealed that SlCCD1A was more closely linked to carotenoid metabolism than SlCCD1B. The overexpression of SlCCD1A indicated that it could cleave lycopene, α-carotene, and β-carotene to produce 6-methyl-5-hepten-2-one, geranylacetone, α-ionone, and β-ionone, increasing the floral, fruity, fatty, and sweet-like aromas of tomato fruits. The flavor quality of tomato fruits could be improved by overexpressing SlCCD1A.

Microstructural, Volatile Compounds, Microbiological and Organoleptical Characteristics of Low-Fat Buffalo Milk Yogurt Enriched with Whey Protein Concentrate and Ca-Caseinate during Cold Storage

Yogurt is a popular fermented milk product across the world. Structure, taste, and odor characteristics are created by fermenting yogurt with diverse ingredients and lactic acid bacteria (LAB), which contribute the most to the acceptance and quality of yogurt. In this study, low-fat buffalo milk yogurts (LFBY) were produced with the enrichment of 1% (w/w) whey protein concentrate (WPC) and Ca-caseinate (Ca-CN). Yogurts were analyzed based on microstructural, microbiological, organoleptical properties; volatile compounds (solid-phase microextraction method associated with gas chromatography-mass spectrometry) during cold storage for 21 days. Yogurts enriched with WPC and Ca-CN had higher total solids, total protein contents, and pH values. A total of 36 volatile components were identified in all produced yogurts. Acetic acid, butanoic acid, acetaldehyde, acetoin, 2,3-butanedione, ethanol, and 1-heptanol were found in significant amounts and mainly contributed to organoleptical properties. Interestingly, the focused volatile compounds that improve taste and odor were higher in LFBY+WPC yogurt than in plain LFBY or LFBY+Ca-CN yogurt. The Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus viable counts were higher in LFBY+WPC yogurt than plain LFBY or/and LFBY+Ca-CN. Conclusively, yogurt enriched with 1% WPC exhibited the best organoleptical properties and volatile component concentrations. The microstructure of the LFBY with WPC was less compact and dense, and regular, with tiny pores and long and individualized casein filaments than the other treatments. The microstructure of the Ca-CN samples caused a compact structure and coarse than in the control yogurt.

Investigation of the Occurrence of Cyanotoxins in Lake Karaoun (Lebanon) by Mass Spectrometry, Bioassays and Molecular Methods

Lake Karaoun is the largest artificial lake in Lebanon and serves multiple purposes. Recently, intensive cyanobacterial blooms have been reported in the lake, raising safety and aesthetic concerns related to the presence of cyanotoxins and cyanobacterial taste and odor (T&O) compounds, respectively. Here, we communicate for the first time results from a recent investigation by LC-MS/MS covering multiple cyanotoxins (microcystins (MCs), anatoxin-a, cylindrospermopsin, nodularin) in water and fish collected between 2019 and 2020. Eleven MCs were identified reaching concentrations of 211 and 199 μg/L for MC-LR and MC-YR, respectively. Cylindrospermopsin, anatoxin-a and nodularin were not detected. The determination of the total MCs was also carried out by ELISA and Protein Phosphatase Inhibition Assay yielding comparable results. Molecular detection of cyanobacteria (16S rRNA) and biosynthetic genes of toxins were carried out by qPCR. Untargeted screening analysis by GC-MS showed the presence of T&O compounds, such as β-cyclocitral, β-ionone, nonanal and dimethylsulfides that contribute to unpleasant odors in water. The determination of volatile organic compounds (VOCs) showed the presence of anthropogenic pollutants, mostly dichloromethane and toluene. The findings are important to develop future monitoring schemes in order to assess the risks from cyanobacterial blooms with regard to the lake’s ecosystem and its uses.

Events linked to Geosmin and 2-methylisoborneol (2-MIB) in a Water Supply in the State of Rio de Janeiro, Brazil: a case study

Tastes and odors in tap water are problems faced by water companies worldwide, with consumers complaints mainly during summer, when cyanobacterial blooms occur and produce compounds such as geosmin and 2-methylisoborneol (2-MIB). We analyzed the data on taste and odor intensity and total concentration of geosmin and 2-MIB compounds in drinking water and raw water collected by the sanitation company supplying of the metropolitan region of Rio de Janeiro (Brazil) during the 2020 and 2021 water crises. Statistical and metagenomic analyses of the raw water samples of the year 2020, were performed. Organoleptic data allowed to signal the presence of these taste and odor (T&O) compounds in the drinking water, and the mean values of taste intensity were above the maximum allowed value of the Brazilian legislation, on average 37.5 times in 2020 and 5 times in 2021, indicating that the measures did not eliminate the problem. There was a linear correlation of 0.97 between the standard organoleptic taste and the total concentration of T&O in 2020. Metagenomic data, from raw water in the year 2020, for the mtf, mic and glys genes indicated 2-MIB as responsible for T&O. Modifications in the surveillance system of catchment and drinking water quality need to be adopted to circumvent the problems of cyanobacterial blooms in the Guandu basin, as conditions favorable to blooms will occur as long as the sanitation problems in this watershed are not solved.

MONITORING OF 2-METHYLISOBORNEOL AND GEOSMINE IN A CONSTRUCTED LAKE TO PUBLIC CATCHMENT IN SOUTHERN BRAZIL

People that consumed treated water from Dourado Lake, Santa Cruz do Sul, RS, Brazil perceived alterations in its taste and odor. Based on this, we focused our study on the monitoring of 2-MIB and GSM in samples collected from Dourado Lake, using solid-phase microextraction (SPME) coupled to gas chromatography/mass spectrometry (GC/MS). The monitoring was done by performing exploratory evaluations at several points on the lake during the summers of 2017 and 2018 and in all seasons of 2019, considering points of water in an inlet and an outlet of the lake. At the inlet point, the average concentration of GSM was 7.56 ± 1.94 ng L-1 and that of 2-MIB was 33.09 ± 6.89 ng L-1. However, for the outlet point, the average concentrations of GSM and 2-MIB were 10.62 ± 2.51 ng L-1 and 28.72 ± 10.47 ng L-1, respectively. In all cases, the presence of GSM and 2-MIB was perceptible by the people consuming the water (during all seasons), showing the need for correct management of water resources.

Encapsulation of Active Pharmaceutical Ingredients in Lipid Micro/Nanoparticles for Oral Administration by Spray-Cooling

Nanoencapsulation via spray cooling (also known as spray chilling and spray congealing) has been used with the aim to improve the functionality, solubility, and protection of drugs; as well as to reduce hygroscopicity; to modify taste and odor to enable oral administration; and many times to achieve a controlled release profile. It is a relatively simple technology, it does not require the use of low-cost solvents (mostly associated to toxicological risk), and it can be applied for lipid raw materials as excipients of oral pharmaceutical formulations. The objective of this work was to revise and discuss the advances of spray cooling technology, with a greater emphasis on the development of lipid micro/nanoparticles to the load of active pharmaceutical ingredients for oral administration.