• Korean Journal of Microbiology
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• v.47 no.4
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• pp.302-307
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• 2011

Native chitin deacetylase of Aspergillus nidulans was purified to apparent homogeneity by a combination of phenyl-Sepharose and Q-Sepharose column chromatography. In order to analyze the amino acid residues involved in the enzyme activity, the enzyme was chemically modified with chemical agent, which selectively reacted with the specific amino acid residue on the protein. When the enzyme was chemically modified with diethylpyrocarbonate, which specifically reacted with histidine residues on the protein, the activity was eliminated. The chitin deacetylase, chemically modified with 100 ${\mu}M$ modifier at the residue of arginine or tyrosine, has shown to have decreased activities. It was shown that the modification at aspartic acid or glutamic acid did not affect the enzyme activity to a greater extent, which would not implicate that acid amino residues were directly involved in catalytic reaction and would affect on the global structures of the proteins. This results demonstrated that histidine and tyrosine residues of enzyme would participate in an important function of the chitin deacetylase activity.

• Food Science of Animal Resources
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• v.30 no.1
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• pp.36-41
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• 2010

The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.39-46
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• 2000

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50${\mu}{\textrm}{m}$ $\alpha$-tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100${\mu}{\textrm}{m}$ cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

• BMB Reports
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• v.34 no.6
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• pp.559-565
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• 2001

Protein carboxyl O-methyltransferase (E.C.2.1.1.24) may play a role in the repair of aged protein that is spontaneously incorporated with isoaspartyl residues. The porcine brain carboxyl O-methyltransferase was cloned in the pET32 vector, and overexpressed in E.coh (BL21) that harbors pETPCMT, which encodes 227 amino acids, including tagging proteins at the N-terminus. The protein sequence of the cloned porcine brain PCMT (r-pbPCMT) shares a 98% identity with that of human erythrocyte PCMT and rat brain PCMT. It is 100% identical with that of bovine brain. The r-pbPCMT was purified using Ni-NTA affinity chromatography and digested by enterokinase in order to remove the protein tags. Then Superdex 75HR gel filtration chromatography was performed. The r-pbPCMT exhibited similar in vitro substrate specificities with the PCMT that was purified from porcine brain. The molecular weight of the enzyme was estimated to be 24.5 kDa on the SDS polyacrylamide gel electrophoresis. The $K_m$ value was $1.1{\times}10^{-7}\;M$ for S-adenosyl-L-methionine. S-adnosyl-L-homocysteine was a competitive type of inhibitor with the $K_i$ value of $1.38{\times}10^{-4}\;M$. The enzyme has optimal activity at pH 6.0 and $37^{\circ}C$. These results indicate that the expressed enzyme is functionally similar to the natural protein. It also suggests that it may be a suitable model to further understand the function of the mammalian enzyme.

• Journal of Ginseng Research
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• v.24 no.2
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• pp.94-98
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• 2000

Tetracycline hydrochloride (TC) caused 100%, 50% and 20% mortality rates among rats injected with 40 mg, 30 mg and 20 mg/100g. b.w. respectively; while the morta]ity rates were decreased to 50%, 20% and 10% when Panax ginseng (2 mg/100g. b.w.) injected with TC during 72 hrs. post-injection. Subacute-toxicity study demonstrated that TC caused severe hepato-nephrotoxicity (demonstrated by biochemical analysis of serum including: transferases , alkaline phosphatase, total protein, glucose, cholesterol urea and creatinine) in rats injected i.p. with 10 mg and 5 mg/100g. b.w. for 7 days of daily injection . These signes of toxicity were greatly diminished by P. ginseng addition to TC doses.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.2
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• pp.247-255
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• 2009

This study examined the quality characteristics of brown sauce prepared with rice powder in place of wheat flour, which is the commonly used thickener. With regard to the general components, as the level of rice powder increased and its ratio in the stock increased, moisture decreased and crude protein, lipid, and ash increased. Reducing sugar content was 4.72% in the control group, and as the mixing ratio of stock to rice powder increased from 100:5 (S1) to 100:13 (S5) in the rice powder-added groups, reducing sugar content increased from 3.94% to 4.82%, respectively. In terms of Hunter's color values, as more rice power was added to the beurre mani$\acute{e}$, L, a, and b values increased. Thus, a sauce of a light brown color was appropriately made. For consistency, as more rice powder beurre mani? was inserted into the analyzer, flow was reduced and viscosity significantly increased. The pH of the control (CS) was 4.94, and there were no significant differences between the control and the rice powder-added groups. In sensory evaluations, color presented its highest score as 6.47 when the mixing ratio of stock to beurre mani$\acute{e}$ was 100:7 (S2). S2 also received the highest score for flavor at 6.32. Viscosity was highest in the control (CS) at 6.26, and was not significantly different from the 100:7 (S2) or 100:9 (S3) groups. Finally, the S2 group had the highest scores for taste (6.21) and overall-acceptability (6.30). In correlation analyses between the sensory characteristics, overall-acceptability as well as the color, flavor, viscosity, and taste of the brown sauce all presented comparatively high positive correlations, in which overall-acceptability and color had the highest correlation.

• Korean Journal of Plant Taxonomy
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• v.53 no.3
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• pp.237-241
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• 2023

The genus Scrophularia L. (Scrophulariaceae) comprises 200-270 species worldwide and is a taxonomically challenging lineage, displaying morphological diversity and hybridization. S. kakudensis is morphologically similar to the closely related taxa S. kakudensis var. microphylla, S. pilosa, and S. cephalantha. Therefore, the purpose of this study was to sequence the chloroplast (cp) genome of S. kakudensis using next-generation sequencing and compare it to those of related taxa. The complete cp genome sequence of Scrophularia kakudensis was found to be 152,355 bp long, consisting of a pair of inverted repeats of 25,485 bp that separate a large single-copy (LSC) of 83,479 bp from small single-copy regions of 17,909 bp. The cp genome contained 78 protein-coding genes, 30 tRNAs, and four rRNAs. A phylogenetic analysis based on 78 protein-coding genes from six Scrophularia species showed S. kakudensis and S. cephalantha formed with 100% bootstrap values. We compared the complete cp genomes of S. kakudensis and S. cephalantha and identified seven sequence divergence regions: matK/rps16, rps16/trnQ, trnS/trnG, rpoB/trnC, trnS/trnG, rpl32/trnL, and ndhD/psaC. These regions may be useful for determining the phylogenetic relationships among S. kakudensis-related species.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.275-282
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• 1998

To investigate the increased acid tolerance of a acid-resistant mutant Leuconostoc paramesenteroides P-100 improved as a kimchi starter, proton permeability, ATPase acitivity, glycolysis activity, $Mg^2$sup +/ releasement, and membrane fatty acid composition were studied and comprised with its wild type Leuconostoc paramesenteroides Pw. In the proton permeability experiment, the maximum values of the average half time (t$\_$1/2/) of pH equilibration through the cell membrane of the Pw and the P-100 were about 6.4 min and 7.8 min in 150 mM KCI solution, respectively. In the 3% NaCl solution, the t$\_$1/2/ values of the Pw and the P-100 were 5.5 min and 6.9 min, respectively. The values and pHs of maximal specific activities of ATPase originated from the Pw and the P-100 were 0.5 unit/mg protein and 0.78 unit/mg protein at pH 6.0, respectively. The result of pH dependence of glycolysis showed that the P-100 had higher activities than that of Pw except at pH 7.0. The releases of magnesium from the Pw and the P-100 were observed about 54.5% and 23.2% at pH 4.0 after 2 hours, respectively. The results of comparison of membrane fatty acid composition of the Pw with the P-100 showed that C$\_$8:0/, C$\_$9:0/, C$\_$10:0/, C$\_$11:0/, C$\_$18:0/, and C$\_$19:0,cyclo/ were major different fatty acids between two strains and the content of C$\_$18:1/, and C$\_$19:0,cyclo/ were 2.8%, N.D (not detected) in the Pw and 0.4%, 2.3% in the P-100. These results indicated that acid tolerance of the P-100 was significantly improved in comparison with its wild type Pw.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.8
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• pp.1205-1214
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• 1999

Whole lupinus albus seeds were pressure toasted at temperatures of 100, 118 and $136^{\circ}C$ for 3, 7, 15 and 30 min to study rumen degradation and post-rumen digestion and to determine optimal heating conditions for the Dutch dairy feed industry. In sacco nylon bag and mobile bag techniques were employed for rumen and intestine incubations to determine ruminal degradation characteristics and intestinal digestion of crude protein (CP) in 4 lactation rumen cannulated and 4 lactating intestinal cannulated Dutch dairy cows fed 47% hay and 53% concentrate according to Dutch dairy requirements. Measured rumen degradation characteristics were soluble fraction (S), undegradable fraction (U), potentially degradable fraction (D), lag time (T0) and rate of degradation (Kd) of insoluble but degradable fraction. Percentage bypass feed protein (BCP), ruminal microbial protein synthesized based on available nitrogen (N_MP) and that based on available energy (E_MP), true protein supplied to the small intestine (TPSI), truly absorbed BCP (ABCP), absorbed microbial protein (AVP) in the small intestine, endogenous protein losses in the digestion (ENDP), true digested protein in the small intestine (TAP or DVE in Dutch) and degraded protein balance (PDB or OEB in Dutch) were totally evaluated using the new Dutch DVE/OEB System. Pressure toasting decreased (p<0.001) rumen degradability of CP. It reduced S (p<0.05) and Kd (p=0.06), increased D (p<0.05) and U (p<0.01) but did not alter T0 (p>0.05), thus resulting in dramatically increased BCP (p<0.001) with increasing time and temperature from 73.7 (raw) up to 182.5 g/kg DM ($136^{\circ}C/15min$). Although rumen microbial protein synthesized based on available energy (E_MP) was reduced, true protein (microbial and bypass feed protein) supplied to the small intestine (TPSI) was increased (p<0.001) from 153.1 (raw) to 247.6 g/kg DM ($136^{\circ}C/15min$). Due to digestibility of BCP in the intestine not changing (p>0.05) average 87.8%, the absorbed BCP increased (p<0.001) from 62.3 (raw) to 153.7 g/kg DM ($136^{\circ}C/15min$). Therefore DVE value of true digested protein in the small intestine was significantly increased (p<0.001) from 118.9 (raw) to 197.0 g/kg DM ($136^{\circ}C/15min$) and OEB value of degraded protein balance was significantly reduced (p<0.001) from 147.2 (raw) to 63.1 g/kg DM ($136^{\circ}C/15min$). It was concluded that pressure toasting was effective in shifting degradation of CP of lupinus albus from the rumen to small intestine without changing intestinal digestion. Further studies are required on the degradation and digestion of individual amino acids and on the damaging effects of processing on amino acids, especially the first limiting amino acids.

• Mycobiology
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• v.43 no.2
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• pp.157-162
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• 2015

Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.