A Comparative Biochemical Characterization of Microbial Transglutaminases: Commercial vs. a Newly Isolated Enzyme from <Emphasis Type="Italic">Streptomyces</Emphasis> Sp.

A new microbial transglutaminase (MTGase or MTG, EC 2.3.2.13) from a Streptomyces sp. strain isolated from Brazilian soil samples was characterized in crude and purified forms. The aim of this work is to provide relevant information about a new transglutaminase and to compare its characteristics with the well-known commercial transglutaminase from Ajinomoto Co. Inc. (Activa® TG-BP). The enzyme from Streptomyces sp., in both crude and pure forms, exhibited optimal activity in the 6.0–6.5 pH range and at 35–40°C. The results for the commercial enzyme were the same. A second maximum of activity was observed at pH 10.0 with both the crude Streptomyces sp. enzyme and the commercial enzyme. This interesting fact has not been reported in the literature previously. The fact that this second maximum of activity does not appear on the purified form of the enzyme may suggest the presence of an isoenzyme on the crude extract. All of the enzymes tested were stable over the pH range from 4.5 to 8.0 and up to 45°C. The decline in activity of the commercial transglutaminase above 45°C and pH 8.0 was more gradual. The activities of all the MTG samples were independent of Ca⁺² concentration, but they were elevated in the presence of K⁺, Ba²⁺, and Co²⁺ and inhibited by Cu²⁺ and Hg²⁺, which suggests the presence of a thiol group in the MTG’s active site. The purified enzyme presented a K _m of 6.37 mM and a V _max of 1.7 U/mL, while the crude enzyme demonstrated a K _m of 6.52 mM and a V _max of 1.35 U/mL.     

Vitamins in brewing: presence and influence of thiamine and riboflavin on wort fermentation

Thiamine and riboflavin vitamers are present in a wide range of foods including beer. These vitamers play critical roles in a variety of enzymatic complexes and can promote and maintain metabolism. Currently, the presence and role of these vitamers in the malting and brewing industry have not been widely explored. This research investigated the effects of various fermentation conditions that may lead to the variations in the vitamin content in beer observed by previous researchers. The present research found that during fermentation, the thiamine content of wort is quickly utilized within the first 6 h of a standard fermentation and the uptake of this vitamin is not affected by increases in wort gravity. While no significant changes were observed in extracellular phosphorylated vitamers of thiamine, both free thiamine and thiamine diphosphate accumulated intracellularly during the wort fermentation. Meanwhile extracellular riboflavin vitamers were only poorly utilized during beer fermentations, however flavin mononucleotide rapidly accumulated intracellularly and more so under aerobic conditions. When yeast was exposed to an all‐malt high‐gravity wort, the thiamine or riboflavin utilization was not affected. However, thiamine utilization was reduced in adjunct‐driven high‐gravity worts. Notwithstanding the lowered thiamine uptake under high‐gravity conditions; there were some minor improvements in fermentation performance and yeast viability. The addition of thiamine to an all‐malt wort did appear to enhance yeast viability, both under normal and high‐gravity conditions. Copyright © 2016 The Institute of Brewing & Distilling     

Vitamins in brewing: effects of post‐fermentation treatments and exposure and maturation on the thiamine and riboflavin vitamer content of beer

Post‐fermentation processes and maturation are important steps in beer production as they help to shape the organoleptic properties and stabilize the final product. Brewers can use a variety of processing aids (e.g. isinglass, PVPP, etc.) and processes (e.g. pasteurization, bottle conditioning, etc.) to achieve a desired final product with a desirable shelf life; however, these processes can have detrimental effects on the vitamin content of the beer. This research found that heat treatments have a marked influence on the decrease in the thiamine diphosphate vitamer, while PVPP and silica treatments have a greater influence on the decrease in riboflavin vitamers. Refrigeration, filtration or centrifugation have no, or only very limited, influence on thiamine or riboflavin vitamers, while application of isinglass, bentonite, tannic acid and SO₂ causes a decrease in both thiamine and riboflavin vitamers. Storage of beer at refrigerated temperatures appears to provide protection against significant degradation of both thiamine and riboflavin vitamers; however, storage of filtered beer at elevated temperatures shows a decrease in thiamine diphosphate and riboflavin. Storage of bottle‐conditioned beer at elevated temperatures shows a marked decrease in yeast viability, accompanied by a decrease in thiamine diphosphate and free riboflavin, and a marked increase in free thiamine. These findings provide an insight into the reason why there is a significant variation in the vitamer content of beers, even within a single beer style. Copyright © 2016 The Institute of Brewing & Distilling     

Shelf-life Assessment of Food Undergoing Oxidation–A Review

Oxidation is the most common event leading to the end of shelf life of microbiologically stable foods. Thus, a reliable shelf-life assessment is crucial to verify how long the product will last before it becomes oxidized to an unacceptable level to the consumers.

Shelf-life assessment strategies of foods and beverages suffering oxidation are critically discussed focusing on definition of the acceptability limit, as well as the choice of the proper oxidative indicators, and methodologies for shelf-life testing. Testing methodologies for shelf-life determination under actual and accelerated storage conditions are considered, highlighting possible uncertainties, pitfalls, and future research needs.     

Dispersive Liquid–Liquid Microextraction Followed by Capillary High-Performance Liquid Chromatography for the Determination of Six Sulfonylurea Herbicides in Fruit Juices

In this study a simple, rapid, and efficient method has been developed for the determination of six sulfonylurea herbicides (SUHs): triasulfuron, metsulfuron-methyl, chlorsulfuron, flazasulfuron, chlorimuron-ethyl, and primisulfuron-methyl in commercial grape and apple juice samples, using dispersive liquid–liquid microextraction coupled with capillary high-performance liquid chromatography with diode array detection. Various parameters that influence the extraction efficiency, such as the type and volume of extraction and disperser solvents, sample pH, and salt addition, were investigated and optimized. Under the optimum conditions, limits of detection and quantification of the method were in the ranges of 2–9 and 8–29 μg L^?1, respectively, lower than the maximum residue limits set by the European Union for the raw fruits, such as grape and apple. The intra- and inter-day relative standard deviations varied from 1.0 to 8.2 and 1.8 to 9.8 %, respectively, with recoveries between 72.0 and 109.5 % for commercial grape (both white and red) and apple juice samples, showing satisfactory accuracy for the determination of SUHs in fruit juices.