Seventeen extracellular protein producing bacteria were isolated from soil samples, among which T219 strain having a strong capability of producing the protein was selected and identified for investigation of biological characteristics. The factors which affect the protein production were investigated and the results are summarized as follows. T219 strain which produces the most extracellular protein was identified as Bacillus sp. Optimum temperature and pH for production of the extracellular protein by T219 strain were $25^{\circ}C$ and 7.5 respectively. Almost no activities of protease and amylase were observed in the protein produced by the protein producing bacteria. In the medium containing yeast extract, the cell growth was moderately high, but almost no accumulation of protein was observed. However, polypeptone had significant effects on both the cell growth and the protein accumulation. The addition of glycine and L-isoleucine to the medium containing polypeptone, yeast extract and meat extract had a great effect on the protein production; 4mg/ml of protein accumulation was observed.
Rashidan, Kianoush Khajeh;Nassoury, Nasha;Giannopoulos, Paresa N.;Guertin, Claude
BMB Reports
/
v.35
no.6
/
pp.595-603
/
2002
A gene that encodes a homologue to baculoviral ODVP-6E/ODV-E56, a baculoviral envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). The ChfuGV odvp-6e/odv-e56 gene was located on an 11-kb BamHI subgenomic fragment using different sets of degenerated primers, which were designed using the results of the protein sequencing of a major 39 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1062 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 353 amino acids with a predicated molecular mass of 38.5 kDa. The amino acid sequence data that was derived from the nucleotide sequence in ChfuGV was compared to those of other baculoviruses. ChfuGV ODVP-6E/ODV-E56, along with othe baculoviral ODVP-6E/ODV-E56 proteins, all contained two putative transmembrane domains at their C-terminus. Several putative N-and O-glycosylation, N-myristoylation, and phosphorylation sites were detected in the ChfuGV ODVP-6E/ODV-E56 protein. A similar pattern was detected when a hydrophobicity-plots comparison was performed on ChfuGV ODVP-6E/ODV-E56 with other baculoviral homologue proteins. At the nucleotide level, a late promoter motif (GTAAG) was located at -14 nt upstream to the start codon of the GhfuGV odvp-6e/odv-e56 gene. a slight variant of the polyadenylation signal, AATAAT, was detected at the position +10 nt that is downstream from the termination signal. A phylogenetic tree for baculoviral ODVP-6E/ODV-E56 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV ODVP-6E/ODV-E56 is most closely related to those of Cydia pomonella granulovirus (CpGV) and Plutella xylostella granulovirus (PxGV).
The effects of dietary protein and lysine levels on lactating multiparous sows and litter performance were studied. Sixty-two crossbred multiparous sows ($Landrace{\times}Yorkshire$) were used. Thirty-three and twenty-nine sows were studied in their second parity and third parity respectively. The three dietary treatments were: (1) the control diet containing 15% CP and 0.75% lysine, (2) a diet containing 13% CP and 0.75% lysine (0.60% natural+0.15% synthetic), and (3) a diet containing 13% CP and 0.60% lysine. They were fed twice daily and allowed ad libitum access to food and water throughout a 28 day lactation from parturition until weaning. The results of this experiment showed that body weight and backfat losses of the sows from farrowing to weaning were significantly affected (p<0.01) by reducing dietary protein. Neither average daily feed intake nor weaning to estrus interval of sows were significantly different among treatments. Supplementing lower dietary protein with synthetic lysine could mitigate backfat losses, but could not prevent body weight losses in lactating multiparous sows. A corn-soybean meal diet containing 13% crude protein and 0.60% lysine did not significantly affect litter size and survival rate of weanling piglets compared with the 15% crude protein diet. There was a tendency towards decreased piglet weight at weaning (p<0.10) and reduced daily gain of piglets (p<0.11) when the multiparous sows were fed the 13% protein diet during lactation. We found a severe loss of body weight and backfat when reducing dietary protein for lactating multiparous sows.
Kim, Joo-Hee;Park, Ju-Yeon;Hong, So-Young;Kim, Mi-Kyung
Nutrition Research and Practice
/
v.3
no.3
/
pp.200-207
/
2009
This study examined the effects of com gluten (CG) and its hydrolysate consumptions on weight reduction in rats fed a high-fat diet. Eight-month-old male Sprague-Dawley rats (n=40) were fed a high-fat diet (40% calorie as fat) for 4 weeks. They were then randomly divided into four groups and fed the isocaloric diets with different protein sources for 8 weeks. The protein sources were casein (control group), intact CG (CG group), CG hydrolysate A (CGHA group, 30% of protein as peptides and 70% as free amino acids) and CG hydrolysate P (CGHP group, 93% of protein as peptides and 7% as free amino acids). Body weight gain, adipose tissue weights, nitrogen balance, absorptions of energy, protein and fat, lipid profiles in plasma, liver and feces and hepatic activities of camitine palmitoyl transferase (CPT), fatty acid synthase (FAS), malic enzyme (ME) and glucose-6-phosphate dehydrogenase (G6PDH) were assessed. The CGHA diet had the highest amount of BCAAs, especially leucine, and most of them existed as free amino acid forms. The CGHA group showed significant weight reduction and negative nitrogen balance. Protein absorption and apparent protein digestibility in the CGHA group were significantly lower than those in other groups. Adipose tissue weights were the lowest in the CGHA group. Activity of CPT tended to be higher in the CGHA group than in other groups and those of FAS, ME and G6PDH were significantly lower in the CGHA group than in other groups. In conclusion, the CGHA diet which had relatively high amounts of free amino acids and BCAAs, especially leucine, had a weight reduction effect by lowering adipose tissue weight and the activities of FAS, ME and G6PDH in experimental animals, but it seemed to be a negative result induced by lowering protein absorption, increasing urinary nitrogen excretion and protein catabolism.
Effect of the amount of fertilizer, plant density, and harvesting time on the production of tobacco leaf protein and fresh biomass was investigated. Flue-cured tobacco(M tabacum, L., cv. NC 82) seedlings were transplanted in the field dressed 200kg N per ha at 1$\times$105, 3$\times$105, 5$\times$105, 7$\times$105 plants per ha, and were harvested at the time when 6 and 10 weeks after transplanting, respectively. Harvest at 10 weeks after transplanting increased greatly number of leaves per plant and fresh weight of a plant, Precentage of senescent leaf weight, but significantly decreased fresh weight of a leaf and total protein contents g-1 fresh weight of leaf and stalk over the amount obtained from the harvest at 6 weeks after transplanting. Also, fresh leaf numbers of a plant, fresh weight of a leaf and of a plant, and total protein contents g-1 fresh weight of biomass were more decreased, but percentage of senescent leaf weight were remarkably increased under higher plant density. Therefore, it was seemed that harvesting at 6 weeks after transplanting under 1$\times$105 plant density per ha is more effective for producing higher yield of biomass and protein per plant than 10 weeks harvesting with 7$\times$105 population per ha. A trend was observed that biomass and protein yields per ha are positively correlated with plant population. Biomass yield per ha was the greatest at 7$\times$105 density(80.5t), but the peak of protein yield was at the near of 5$\times$105 population(2454kg as total protein) per ha on the regression curve. It was assumed that if tobacco plants are transplanted under 5$\times$105 plant density at the mid of May, and thereafter harvest at 6 weeks repeatedly during the growing season, it is possible to harvest 2~3 times per year, and to yield more 6.024kg of protein and over 140me1ric tons of fresh biomass ha 1 year 1 statistically in the korea tobacco growing regions.
A phylogenetic comparison of homologous protein can often supplement genetic and biochemical analysis by revealing conserved structures that are critical for function(Waugh et al., 1989). I therefore isolated a secY homologue from B. subtilis, a gram positive bacterium evolutionary distant from E. coli. The comparison and interplay between these two bacterial systems should contribute greatly to our understanding of the functions and interactions within systems evolved for protein translocation in both prokaryotic and eukaryotic organisms.
Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
Journal of Plant Biotechnology
/
v.43
no.2
/
pp.231-239
/
2016
To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.
Yoon, Sun Young;Joo, Jong Hyuck;Kim, Joo Heon;Kang, Ho Bum;Kim, Jin Sook;Lee, Younghee;Kwon, Do Hwan;Kim, Chang Nam;Choe, In Seong;Kim, Jae Wha
IMMUNE NETWORK
/
v.4
no.1
/
pp.23-30
/
2004
Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.
Our nation is confronted with the situation that the rice, a principal food, short of some essential amino acids, leads to imbalanced meals insufficient in the nutrient of Protein, to bring many difficulties in the elevation of nutritional state in our nation. While. our country has been produced much amounts of Panax Ginseng roots which has a stimulating effects on the metabolism of protein, lipid and nucleic acids in the body. And the leaf and trunk of Panax Ginseng were also produced a considerable amounts as the by-products. Author believe that these by-products (leaf and trunk) of Panax Ginseng might have some components possessing simillar activity with Panax Ginseng root although the quantity and qualify of the functional components may more or less be different. Therefore, this study was demised to observe the supplemental effect of the Panax Ginseng-by-Products on the dietary protein efficiency and nutritional state of rats. The feeds used for this experiment were rice containing 30% barely, fish four, and the leaf, trunk and small root of the Panax Ginseng, and the contents of the general nutrients including protein, lipid and carbohydrate etc. in each feed were analyzed for the combination of each feed. And, being based on analytical values of Protein in food. fish Pour as Protein source was added were rice containing 30% barely to be include 8.6 to 8.7%, 12%, 15% and 18% of protein. Then 2% of the leaf, trunk or small reef of Panax Ginseng was supplemented into each of above protein diet group, ton 16 kinds of diets were Prepared. The male albino rats from a Pure strain, weighing 70g to 80g. were used for experimental animals. They were maintained with coresponding fist for f and 8 weeks, and the growth rate, consumption of diets and protein, efficiency of feed and Protein in animals were determined. The lipids, proteins and cholesterols in serum and liver were also determined quantitatively after they were sacrificed in coresponding term. The results obtained are summarized as follows: 1. Body weigh of diet group containing 8.6 to 8.7%,12%, and 15% of protein are increased remarkably by supplement of 2% of the leaf or small root of Panax Ginseng in comparison with each of controls. But this tendency could not observed in diet group containing 18eA Proteins. 2. Feed efficiency showed same tendency in comparison with changes of gained body weight. Specially, in each of diets containing 8.7%, 12%, 15% and 18% of Proteins, supplement of the leaf of Panax Ginseng showed the better feed efficiency than supplement of the trunk or small root. 3. In feeding group for 8 weeks, protein efficiency showed worst efficiency in diet containing 18% proteins and showed the best efficiency was the diet group containing 12% Proteins. And the efficiency was improved according to supplement of the leaf of Panax Ginseng. 4. Nitrogen contents in serum and liver did not show large differences each other in all diet groups. But contexts of total cholesterol and 1ipid were decreased markedly in diet groups containing 12%, 15% and 18% of proteins in comparison with diet group containing 8.6% to 8.8% of proteins.
Journal of the Korean institute of surface engineering
/
v.37
no.2
/
pp.76-79
/
2004
Nickel chloride coated protein chip plate was developed by using a spin coating method. The ability of histidine tagged protein adsorption was investigated at various solvents. The surface of plate has a large aggregated nickel complex with high density in water. However, the surface of plate has a very small size of aggregated nickel complex with low density in isopropanol. The ability of protein adsorption decreased as increasing the size of alkyl chain in various alcohol solvents. The mechanism on the ability of protein adsorption at the plate surface is discussed.
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