BackgroundLipolytic enzymes are commonly used to produce desired flavors in lipolyzed milkfat (LMF) manufacturing processes. However, the choice of enzyme is critical because it determines the final profile of fatty acids released and the consequent flavor of the product. We previously constructed a metagenomic library from marine sediments, to explore the novel enzymes which have unique properties useful in flavor-enhancing LMF.ResultsA novel lipase Est_p6 was isolated from a metagenomic library and was expressed highly in E.coli. Bioinformatic analysis indicated that Est_p6 belongs to lipolytic enzyme family IV, the molecular weight of purified Est_p6 was estimated at 36 kDa by SDS-PAGE. The hydrolytic activity of the enzyme was stable under alkaline condition and the optimal temperature was 50°C. It had a high specific activity (2500 U/mg) toward pNP butyrate (pNP-C4), with Km and Vmax values of 1.148 mM and 3497 μmol∙min-1∙mg-1, respectively. The enzyme activity was enhanced by DTT and was not significantly inhibited by PMSF, EDTA or SDS. This enzyme also showed high hydrolysis specificity for myristate (C14) and palmitate (C16). It seems that Est_p6 has safety for commercial LMF flavor production and food manufacturing processes.ConclusionsThe ocean is a vast and largely unexplored resource for enzymes. According the outstanding alkaline-stability of Est_p6 and it produced myristic acid and palmitic acid more efficiently than other free fatty acids in lipolyzed milkfat. This novel lipase may be used to impart a distinctive and desirable flavor and odor in milkfat flavor production.

Ferric uptake regulator (Fur) plays a key role in the iron homeostasis of prokaryotes, such as bacterial pathogens, but the molecular mechanisms and structural basis of Fur–DNA binding remain incompletely understood. Here, we report high-resolution structures of Magnetospirillum gryphiswaldense MSR-1 Fur in four different states: apo-Fur, holo-Fur, the Fur–feoAB1 operator complex and the Fur–Pseudomonas aeruginosa Fur box complex. Apo-Fur is a transition metal ion-independent dimer whose binding induces profound conformational changes and confers DNA-binding ability. Structural characterization, mutagenesis, biochemistry and in vivo data reveal that Fur recognizes DNA by using a combination of base readout through direct contacts in the major groove and shape readout through recognition of the minor-groove electrostatic potential by lysine. The resulting conformational plasticity enables Fur binding to diverse substrates. Our results provide insights into metal ion activation and substrate recognition by Fur that suggest pathways to engineer magnetotactic bacteria and antipathogenic drugs.

This study is underpowered, but there are no trends to suggest that d-MPH, taken concurrently with adjuvant chemotherapy, improves quality of life or fatigue.

Dietary threonine imbalance is known to reduce the growth of the small intestine, liver, and skeletal muscle in young animals, but the underlying mechanism is largely unknown. Using the pig model, this study was conducted to test the hypothesis that either a deficiency or an excess of dietary threonine impairs protein synthesis in these tissues. Young pigs (25 d of age) were fed diets containing 0.37, 0.74 (current NRC requirement) or 1.11% true ileal digestible threonine (TIDT) (n = 6/diet). Pigs receiving the 0.74 and 1.11% TIDT diets were pair-fed with the same amount of feed as pigs receiving the 0.37% TIDT diet. After a 14-d dietary treatment, the fractional synthesis rate (FSR) of protein in tissues was measured using a flooding dose of l-phenylalanine plus L-[ring-(2)H(5)]phenylalanine. The results indicated that the FSR of protein in liver was reduced (P < 0.05) in pigs fed the 0.37% TIDT diet compared with pigs fed the 0.74 or 1.11% TIDT diet, and did not differ between pigs fed the 0.74 and 1.11% TIDT diets. The FSR of protein in longissimus muscle, jejunal mucosa, and mucins was reduced (P < 0.05) in pigs fed the 0.37 or 1.11% TIDT diet compared with pigs fed the 0.74% TIDT diet. The absolute synthesis rate of protein in the jejunal mucosa and muscle was also reduced (P < 0.01) in pigs fed the 0.37 and 1.11% TIDT diets compared with the controls. The absolute synthesis rate of hepatic protein was lower (P < 0.01) in pigs fed the 0.37% TIDT diets when compared with pigs fed the 0.74% TIDT diet. Protein synthesis in skeletal muscle as well as jejunal mucosa and mucins was reduced to a greater extent than that in liver in response to an imbalance of dietary threonine. Collectively, these results indicate that either an excess or a deficiency of dietary threonine decreases protein synthesis in rapidly growing tissues of young pigs. The findings provide a mechanism for the low growth performance of animals fed a threonine-imbalanced diet.