To gather the basic data for increasing the utilization of Isatis tinctoria, we examined the effects of both antioxidative enzyme activity and antimicrobial activity from the extract of Isatis tinctoria. Ascorbate Peroxidase activities reveal that there is an decrease in order; ethanol extract from its stem (1601.7 Unit/mg protein), methanol extract from its leaf (1133.7 Unit/mg protein) and distilled water extract from its leaf (524.3 Unit/mg protein). Catalase activities reveal that there is an decrease in order; ethanol extract from its flower petal (177.1 Unit/mg protein), methanol extract from its leaf (120.8 Unit/mg protein) and distilled water extract from its flower petal (55.4 Unit/mg protein). Peroxidase activities reveal that there is an decrease in order; ethanol extract from its flower petal (27.1 Unit/mg protein), methanol extract from its flower petal (14.6 Unit/mg protein) and distilled water extract from its stem (10.4 Unit/mg protein). Superoxide dismutase activities reveal that there is an increase in order; distilled water extract from its root (90.8%), methanol extract from its flower petal (80.1%) and ethanol extract from its root (75.5%). Its flower extract showed a antimicrobial activity only against Vibrio parahaemolyticus, its root extract had only against Staphylococcus aureus, and its stem extract had against Bacillus subtilis, Escherichia coli and Staphylococcus aureus, regardless of solvents. Especially, distilled water extract from its leaf showed a high antimicrobial activity against both Bacillus subtilis and Escherichia coli and inhibition diameters against those were 30.0 and 24.0 mm, respectively.
Porphyromonas gingivalis, a Gram negative, anaerobic, asaccharolytic rod, is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is both hemin and iron regulated has recently been purified and characterized in this oral pathogen. This study has identified a hemin-binding P. gingivalis protein by expression of a P. gingivalis genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. A library of genomic DNA fragments from P. gingivalis was constructed in plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$ , and screened for recombinant clones with hemin-binding activity by plating onto hemin-containing agar. Of approximately 10,000 recombinant E. coli colonies screened on LB-amp-hemin agar, 10 exhibited a clearly pigmented phenotype. Each clone contained various insert DNA. The Hind III fragment transferred to the T7 RNA polymerase/promoter expression vector system produced a sligltly smaller (21 kDa) protein, a precursor form, immunoreactive to the antibody against the 24 kDa protein, suggesting that the cloned DNA fragment probably carried an entire gene for the 24 kDa hemin-binding protein.
This experiment was conducted to investigate the effects of different levels of dried leftover food (DLF) in the diet on feed utilization and egg-laying performance of hens. One hundred sixty-eight, 18 week old Tetra brown commercial layers, were assigned to 7 treatments in a completely randomized design. Each treatment has four replications per treatment with six animals per replication. All the experimental animals were fed diets for 7 weeks. The treatments included 1) control group without DLF, 2) diet with 10% DLF, 3) diet with 20% DLF, 4) diet with 30% DLF, 5) 10% higher protein level of diet with 10% DLF, 6) 20% higher protein level of diet with 20% DLF and 7) 30% higher protein level of diet with 30% DLF. Average daily feed intake (ADFI) tended to be improved with DLF feeding. ADFI of group fed diets with 20% was significantly higher than that of control (p<0.05). Feed conversions of DLFfed groups were higher than that of control. Egg production tended to be higher in groups fed diets with 10% DLF than control diet without significant differences (p>0.05). However, those of groups fed diets containing 20 and 30% DLF were lower than that of control. Supplementing protein source to DLF-containing diets improved egg production (p<0.05). Increasing level of DLF in the diet for layer decreased egg weight and egg mass compared to control without significant differences (p>0.05). Protein supplementation to DLF-containing diets increased egg mass without significant difference (p>0.05). The range of egg cholesterol concentration of DLF-fed groups was 11.94-14.10 mg/g while that of control group was 12.31 mg/g although there was no significant difference among treatments (p>0.05).
Defatted sesame flour is the by-products obtained after oil extracting process. Although this flour has high quality and quantity of protein its use is limited only for animal feed and fertilization. Sesame seeds contain antinutrients such as oxalate, phytate and phenol compounds and these compounds lower their nutritive value. recently, ultrafiltration(UF) has been used to concentrate protein from various food sources. This study was carried out to examine the effects of UF with different membrane pore size on the components of sesame protein concentrates including antinutrients and to compare with that of conventional acid-precipitated sesame protein isolate. The protein contents of sesame protein concentrates prepared by JF using 10K, 30K, 100K were 84.2%, 82.7%, 76.4% and the protein yields were 36.44%, 34.69, 31.43% and the protein contents was 88.7% Alkali extraction process at pH 9.0 followed by UF technique reduced oxalate and phytate content. There were 85% and 94% reduction of oxalate and phytate content by UF with membrane pore size of 100K daltons, respectively. However, the content of total phenol compounds was not reduced by this method. About 99% of calcium and 50% of zinc were removed by UF with membrane of 100K daltons. total essential amino acid contents of sesame protein concentrates prepared by UF were decreased slightly when compared with acid-precipitated sesame protein concentrate.
Nef is a lentiviral protein involved in pathogenesis of AIDS, but its molecular mechanism of action remains incompletely understood. Here we report the isolation of the interacting protein with the HIV-1 Nef, using the yeast two hybrid system for expression cloning. One of the positive colonies was selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed that this interacting protein is Human Ikaros/LyF-1. This protein interacted with the C-terminal region of Nef specifically in yeast system, not with the N-terminal region. This interaction was also confirmed by in vitro binding assay.
In order to investigate the effect of dietary zinc and phytic acid levels on protein metabolism in rats, male rats of Sprague-Dawley strains weighing approximately $60\~74g$ were fed different diets which contained 0, 0.35 and $1.05\%$ phytic acid each at 3 levels of zinc(0, 30 and 1,500 ppm zinc) for 28 days. Result obtained in this experiment are summarized as follows; 1. Body weight gait food consumption food efficiency ratio and protein efficiency ratio were lower in the rats fed zinc deficient diet(0 ppm zinc) than in those consuming 30 or 1,500 ppm dietary zinc, and the additional effect of phytic acid were not observed in all of then 2. Liver weight was lower in the rats fed 30 ppm zinc diet than in those fed 0 or 1,500 ppm-zinc diet but kidney and spleen weights were lower in the rats fed zinc deficient diet than in those fed 30 or 1,500 ppm-zinc diet Among organs measured only the liver appeared to be influenced by dietary phytic acid: the more the dietary phytic acid, the more the weight of liver, 3. Fecal nitrogen was decreased in the rats fed zinc deficient diet compared with those fed 30 or 1,500 ppm dietary zinc. Urinary nitrogen was increased in the rats fed $1.05\%$ dietary phytic acid compared with those fed 0.35 or $0\%$ dietary phytic acid Nitrogen retention of rat was influenced by neither dietary zinc nor phytic acid. 4. Urea nitrogen was decreased with increasing dietary zinc levels, and creatinine and uric acid levels were increased with increasing dietary zinc concentration or with additional quantity of phytic acid. Uric acid appeared to be influenced by zinc x phytic acid interaction; especially, the presence of phytic acid in the 30 ppm-zinc diet had significant effect on uric acid content. 5. Hemoglobin concentrations and hematocrit ratio were higher in the rats fed 30 ppm dietary zinc than in those fed 0 or 1,500 ppm-zinc diet Serum zinc concentration was increased with increasing dietary zinc levels. The content of total protein albumin and BUN and the ratio of albumin to globulin in serum, and protein content in liver were influenced by neither dietary zinc nor phytic acid.
Ha, Jung-Hye;Jeong, Mi-Suk;Jo, Wol-Soon;Jeong, Min-Ho;Jang, Se-Bok
Bulletin of the Korean Chemical Society
/
v.31
no.2
/
pp.275-280
/
2010
Actinobacillus actinomycetemcomitans is a gram-negative, nonmotile coccobacillus bacterium that is associated with several human diseases, including endocarditis, meningitis, osteomyelitis, subcutaneous abscesses and periodontal diseases. A full-length Omp1-like protein gene from A. actinomycetemcomitans was cloned into a pQE30 vector and overexpressed in Escherichia coli BL21(DE3) cells. The protein revealed sequence homologies to Seventeen kilodalton proteins (Skp) from Pasteurella multocida and E. coli that have been characterized as periplasmic chaperones. This soluble Omp1-like protein was successfully purified to homogeneity for further folding and functional studies. The purity, identity, and conformation of the protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, fluorescence spectroscopic, and differential scanning calorimetric studies. We showed that the protein formed an oligomer larger than a tetramer. We found, further, that it is comprised of mostly $\alpha$-helices and boasts high thermal stability.
This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.
The experiment was conducted to determine the effects of cutting interval and cutting height on the yield and nutrient composition of hedge lucerne (Desmanthus virgatus) when grown on a sandy soil in the Northeast of Thailand. The cutting intervals compared were 30, 40 and 50 days between harvests and the cutting heights 30, 40 and 50 cm above ground level. The experiment was a $3{\times}3$ factorial layout in a randomized complete block design with 4 replications-giving a total of 36 plots each $3{\times}3m^2$. Harvested plant material was weighed, dried and the ground subsamples taken for analyses of crude protein (CP), crude fiber (CF), ash, ether extract (EE) and nitrogen-free extract (NFE). At the last harvest the hedge lucerne samples were separated to determine leaf to stem ratios and then analyzed for nutrient composition in the leaf and stem. Results showed that increasing the cutting interval (i.e. advancing age of maturity) increased dry matter and nutrient yields significantly. In terms of nutrient content, it also increased the crude fiber, ash, ether extract and nitrogen free extract percent in the plant. However, crude protein percent was markedly decreased as the cutting interval increased. Increasing cutting height had no effect on dry matter yield and yields of nutrients, but in terms of nutrient content, it increased crude protein and ash content, but decreased crude fiber content. The percent EE and NFE in the plant was unaffected by cutting height. From the results presented it is clear that cutting a stand of hedge lucerne every 40 to 50 days will achieve greater dry matter and nutrient yields than cutting more frequently, at 30 days. The cutting height at harvest, whether 30, 40 or 50 cm above ground level had no effect on dry matter or nutrient yields of hedge Lucerne. Hedge lucerne therefore offers the Thai poultry farmer a useful alternative protein supplement for poultry diets rather than relying on the more expensive soybean meal. As it can be readily and successfully grown on a range of soil types and climates throughout Thailand, hedge lucerne also offers the Thai farmer a valuable additional source of income.
An experiment was conducted to examine the effect of a high environmental temperature on the performance, conformation, and carcass characteristics of pigs and the influence of diet. Thirty-six three-way crossed castrated male pigs with average initial body weight of 50.4 kg were used in the experiment. The pig were allocated to the following treatments: two environmental temperatures (20 and $30^{\circ}C$)${\times}$three dietary energy levels (2,980, 3,300 and 3,600 kcal/kg)${\times}$three protein levels (12.8, 15.2 and 17.2%). Daily weight gain was lighter (p<0.01) and feed: gain ratio lower (p<0.05) in pigs at $30^{\circ}C$ than for pigs at $20^{\circ}C$ The pigs at $30^{\circ}C$ were significantly taller with deeper bodies (p<0.05) and significantly longer (p<0.05) both vertically horizontally in the planum nasal when kept at $30^{\circ}C$. The width of body and the circumference of the neck were greater (p<0.05) at $20^{\circ}C$. The lean meat of the loin, middle section, ham, and hind section were significantly greater (p<0.05) in pigs kept at $30^{\circ}C$ and the belly was significantly heavier. The total unsaturated free fatty acids were significantly higher (p<0.05) in the body fat of pigs kept at $20^{\circ}C$ than in that of pigs at $30^{\circ}C$. The results indicated that when pigs are given very restricted same amounts of feed, they may need less energy to maintain their body temperature under moderately high environmental temperature ($30^{\circ}C$); consequently, their performance is better than that of pigs under optimum environmental temperature. The results showed very clearly that weight gain of pigs increased with increasing of dietary protein and energy content. The increasing of dietary protein content seemed more significant when content increasing to 17.2% compared with the 12.8 and 15.2% protein content treatments. The increasing of dietary energy content was more significant when content increasing to 3,600 compared with the other low energy content treatments.
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