RESUMO
Kombucha is a potential probiotic tea-based drink with increasing worldwide consumption. Studies on this probiotic beverage are growing rapidly, especially about micronutrients and microbial population. As such, the present study performed the molecular identification of the microorganism and evaluated 5-methyltetrahydrofolate content by HPLC-DAD, phenolic compounds, flavonoids, carotenoids, antioxidant activity by spectrophotometric methods, and physicochemical composition of green tea kombucha on fermentation days 1, 3, 7, 14, and 21. DNA sequencing identified the Microbacterium genus as predominant. However, was unable to safely determine the species level because of the rRNA 16S gene sequence similarity between four species M. ureisolvens, M. yannicii, M. chocolatum e M. atlanticum. The concentration of 5-methyltetrahydrofolate found on the third day was 39.12 ± 1.32 µg/mL (liquid) and 45.78 ± 8.42 µg/mL (polymeric biofilm); On the twenty-first day it was 50.87 ± 3.56 µg/mL (liquid) and 54.88 ± 3.89 µg/mL (polymeric biofilm). Total phenolic compounds increased with fermentation; however, flavonoids and carotenoids were degraded by the process. The information on 5-methyltetrahydrofolate is unprecedented and highly relevant for food guidelines, since related deficiencies can lead to fetal malformation in the first three months of pregnancy. Lastly, the best fermentation time to obtain 5-methyltetrahydrofolate and others bioactive compounds is between days 7-14. Further analyses are also encouraged to understand the bioavailability of the vitamin.
RESUMO
NADP+-dependent glutamate dehydrogenase (NADP+-Gdh) is the first step in ammonia assimilation pathway in Saccharomyces cerevisiae and the knowledge of its regulation is the key for many biotechnological purposes such as single cell protein production. The regulation of NADP+-Gdh activity in Kluyveromyces marxianus cells was evaluated under different ammonia supply in batch cultivations. The results showed that K. marxianus NADP+-Gdh activity is induced over a narrow range of extracellular ammonia supply, being repressed by both high ammonia concentration and the glutamate formed. This activity is not growth-associated and may function mainly to trace low amounts of ammonia after growth cessation. The results demonstrated that NADP+-Gdh may not be the main enzyme for ammonia assimilation in K. marxianus, as it has been postulated for K. lactis, instead is subjected to the same regulatory mechanism described for S. cerevisiae.
Glutamato desidrogenase dependente de NADP+ (NADP+-Gdh) constitui o primeiro passo enzimático no mecanismo de assimilação de nitrogênio em Saccharomyces cerevisiae e o conhecimento de sua regulação é chave na iniciativa de vários propósitos biotecnológicos, tais como a produção de proteína microbiana. A regulação da atividade NADP+-Gdh em células de Kluyveromyces marxianus foi avaliada a partir de diferentes condições de suprimento de amonia em cultivo em batelada. Os resultados mostraram que a atividade NADP+-Gdh de K. marxianus foi induzida em uma estreita faixa de concentração de amonia no meio, sendo reprimida tanto por altas concentrações deste composto quanto pelo produto glutamato. Esta atividade não está associada ao crescimento celular e deve funcionar principalmente no rastreamento de pequenas quantidades de amonia após a parada do crescimento celular. Isto demonstra que NADP+-Gdh não deve ser a principal enzima de assimilação de amonia em K. marxianus, como tem sido postulado para K. lactis, contudo deve estar submetida ao mesmo mecanismo regulatório encontrado em S. cerevisiae.