• BMB Reports
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• v.32 no.6
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• pp.579-584
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• 1999

Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

• Korean Journal of Microbiology
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• v.9 no.4
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• pp.163-168
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• 1971

Neutral protease which was obtained from a genus of Aspergilli as the crystal form were investigated for their purification and properties. The results of biochemical and enzymatic studies for their purification and properties in this enzyme were as follows. 1) On the wheat media containing 70%-water and $CaCo_{3}$, Aspergilus oryzae S.H.W. 131 is satisfactorily grown under the basic optimum conditions temperature $27^{\circ}C$- $30^{\circ}C$at relative humidity 100% for three days. 2) The enzyme solution extracted with water is successively purified through the passing on column of Asmti-177N for decolorization of it. And ion exchanger such as DEAAE Sphadex A-50 or Shepadex G-100 and fraction collector is necessary for the sepearte treatments of this enzyme. After washing it with organic solvents as aceton-EtOH, etc., it should be dried on the vacuum dryer at $40^{\circ}C$) The protease activity is determined by the amounts of amino acids, tyrosine. 4) The optimum pH of neutral protease is 6.0-8.0. 5) In effectively decomposing with this neutral protease, the optimum temperature is $35^{\circ}C$. 6) It is interesting that the amounts of metal ion affects the activity of neutral protease. For examples, if it were treated with manganic ion, its activity would be more effective than any other that.

• Journal of Applied Biological Chemistry
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• v.44 no.1
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• pp.6-11
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• 2001

An ${\omega}-3$ fatty acid desaturase gene which is involved in de novo synthesis of -Iinolenate was isolated from cDNA library of Perilla frutescens. A cDNA library was constructed with mRNA extracted from perilla seeds of 12 DAF. The cDNA clone consisting of 1317-bp open reading frame encoding 438 amino acids with a relative MW of 50kDa, was isolated and showed 65-83% similarities to other known genes. This cDNA is deduced to encode a plastidal ${\omega}-3$ fatty acid desaturase based on the fact that it has higher homology to plastidal ones than to microsomal ones and its N-terminal sequence shares several characteristics of transit peptides of chloroplast proteins. Southern blot analysis of genomic DNA indicated that more than one gene or alleles for ${\omega}-3$ fatty acid desaturase are present in the genome of perilla. Northern blot analysis showed that the ${\omega}-3$ fatty acid desaturase gene is mainly revealed in early developing seeds and has different expression patterns depending on tissue types compared to the microsomal ones.

• Preventive Nutrition and Food Science
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• v.12 no.2
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• pp.89-94
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• 2007

Nutrients and hormone levels of dried pig placenta were studied. Placentas were freeze-dried (FD), oven-dried at 60 (OD-60), and 90$^{\circ}C$ (OD-90) and then crushed by a blender into small pieces. FD and OD-60 pig placenta had a higher moisture content than did OD-90, with no difference between FD and OD-60. There were no large differences in compositions of crude protein, crude fat, and crude ash of dried placenta among the treatments and the contents of K, Fe, and ${\alpha}$-tocopherol were highest in FD (p<0.05). Glutamine and glycine were the most abundant amino acids in all dried placenta and tyrosine was highly retained in FD placenta, compared with OD (p<0.05). Estradiol was the major sex hormone, followed by progesterone and testosterone in all dried placentas. Antibiotics including amoxicillin, sulfamethazine, tylosin, and chlorotetracyclin were not detected from the pig placentas tested. These results demonstrate that placenta is a good biomaterial with high nutritional quality, and that freeze drying is superior to oven drying for processing pig placenta.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.413-418
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• 2012

The purpose of this study is to determine the possibility of using Crataegi fructus as a natural food source. To accomplish this purpose, the contents of general and biological activities were measured. The contents of carbohydrate, crude protein, crude lipid and ash are 85.6%, 2.4%, 1.9% and 0.4%, respeectively. Further, the calories of Crataegi fructus was 369.1 kcal. The contents of essential and non-essential amino acids were 852.26 mg and 1,178.29 mg, respeectively. The K was the largest mineral followed by Ca, P, Mg, which means Crataegi fructus is an alkali material. Crataegi fructus extracts slightly(17.6~32.8) inhibited ${\alpha}$-glucosidase activity. However, there is no inhibitory activity against ${\alpha}$-amylase. In terms of proteslytic activity, Crataegi fructus extracts showed a strong activity than pancreatin(used as a positive control). These results indicate that Crataegi fructus can be used as a natural resource for material aiding digestion.

• Korean Journal of Food Science and Technology
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• v.33 no.5
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• pp.603-610
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• 2001

The quality characteristics of soy yogurt prepared with different proteolytic enzymes and starter culture were evaluated. In order to facilitate the growth of lactic acid bacteria and subsequent production of lactic acid, soy protein isolate(SPI) was hydrolyzed using three kinds of proteases; one extracted from Aspergillus oryzae, bromelain and ${\alpha}-chymotrypsin$. The cultural systems employed thereafter for lactic fermentations were: 1) Bifidobacterium bifidum, 2) B. bifidum and Lactobacillus acidophilus, 3) B. bifidum and Lactobacillus bulgaricus. In soy yogurt, pH was more decreased by mixed culture method than single culture method with the accumulation of lactic acid. Viable cells of lactic acid bacteria in soy yogurts were measured $10^8$ CFU/g by the single culture method while $10^9$ CFU/g by the mixed culture method except ${\alpha}-chymotrypsin$ treatment. The amount of free amino acids in soy yogurts were substaintially increased by enzyme treatment. Viscosity was decreased by enzyme treatment, resulting in higher viscosity by ${\alpha}-chymotrypsin$ treatment. Water holding capacity was found to be higher in the single culture method in case of enzyme treatment. Among the various volatile flavor components isolated and identified after enzyme hydrolysis, acetaldehyde, ethanol, diacetyl, butyl alcohol contents tended to increase by lactic fermentation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.316-324
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• 1985

Vacuum-packed and seasoned smoked-dried products of red squid, Ommastrephes bartrami, caught in the Northern Pacific Ocean, were prepared and stored at room temperature for 90 days to test their keeping quality. Defrosted squids were eviscerated, skinned, and cut. The mantle meats were flavored with seasoning powders prepared from sugar, sorbitol, salt, monosodium glutamate, or smoke flavor (Smoke-EZ, Alpha Foods Co., Ltd.). After seasoning, the mantle meats were dried at $45^{\circ}C$ for 7 hours, vacuum packed in plastic film bags, and pasteurized in water at $95^{\circ}C$ for 30 minutes. Three kinds of products were prepared : control products (seasoned-dried), solid smoked seasoned-dried and liquid smoked seasoned-dried. The moisture level, water activity, color value (L, a and b value), texture, and viable cell counts of bacteria in these products were determined during storage at room temperature, $5^{\circ}C\;and\;35^{\circ}C$, respectively. The results showed that the products could be preserved at good condition for 90 days though they developed pale brown color during storage. The contents of free amino acids, nucleotides and their related compounds, and the compositions of fatty acids of raw squid and smoked products were analysed. In the amino acids, arginine, taurine, glycine and proline were abundant in raw and smoked products. The contents of hypoxanthine of raw and smoked products were higher than the other nucleotides and their related compounds. In fatty acid compositions of raw and smoked products, the dominant fatty acids were docosahexaenoic acid (22:6), hexadecanoic acid(16:0) and eicosapentaenoic acid (22:5).

• Journal of Microbiology and Biotechnology
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• v.8 no.2
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• pp.152-157
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• 1998

To analyze the role of the C and D domains in the cyclization activity of cyclodextrin glucanotransferase (CGTase), two plasmids, pKB1ΔC300 and pKB1ΔD96, were constructed in which DNA regions encoding 100 and 32 amino acids, respectively, from the C and D domains of B. stearothermophilus NO2 CGTase were deleted. The mutated CGTase from the pKBlΔC300 produced much lower amounts of ${\alpha}$-, ${\beta}$-, and $\gamma$-cyclodextrin (CD) than the parental CGTase. However, the mutated CGTase from the pKBlΔD96 showed a similar production pattern of CDs to wild-type CGTase. The production ratios of the ${\alpha}$-, ${\beta}$- and $\gamma$-CDs were not affected by the deletions, when compared to those of parental CGTase. The optimum temperature of the mutated CGTase from the pKBlΔC300 was decreased from $60^{\circ}C$ to $55^{\circ}C$. The optimum pH of the mutated CGTase from the pKB1D96 was shifted from 6.0 to 7.0. The thermostability of the two mutant CGTases were not changed. From these results, it is suggested that the C and D domains are not related to cyclization activity directly because mutant-enzymes deleted C or D domains still possessed their activity. However, they are important for other enzymatic properties such as productivity and pH optimum as a partition of CGTase tertiary structure.

• Applied Biological Chemistry
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• v.45 no.4
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• pp.203-206
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• 2002

${\gamma}-Tocopherol$ methyltransferase (TMT) is an enzyme catalyzing ${\gamma}-tocopherol$ into ${\alpha}-tocopherol$ at the final step of ${\alpha}-tocopherol$ synthesis pathway. Putative TMT cDNA clone specific to Perilla frutescens immature seeds was isolated from cDNA library. The cDNA clone consisted of 1369 bp open reading frame encoding 369 amino acids with a relative Mw of 42 kDa. Results revealed the CDNA has 60% homology to Arabidopsis thaliana TMT, and possesses methyltransferase and S-adenosyl methionine-binding domains, suggesting that cDNA encodes a ${\gamma}-tocopherol$ methyltransferase To characterize the properties of the TMT gene, the cDNA sequences coding for mature TMT were expressed in E. coli and assayed to determine the enzyme activity in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.615-621
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• 2007

AMP-activated protein kinase alpha 2 (PRKAA2) plays a key role in regulation of fatty acid and cholesterol metabolism. This study investigated the porcine PRKAA2 gene as a positional candidate for intramuscular fat and backfat thickness traits in pig chromosome 6. A partial fragment of the porcine PRKAA2 gene, amplified by PCR, contained a putative intron 3 including a part of exon 3 and 4, comparable with that of human PRKAA2 gene. Within the fragment, several single nucleotide polymorphisms were identified using multiple sequence alignments. Of these, TaqI restriction enzyme polymorphism was used for genotyping various pig breeds including Korean reference family. Using linkage and physical mapping, the porcine PRKAA2 gene was mapped in the region between microsatellite markers SW1881 and SW1680 on chromosome 6. Allele frequencies were quite different among pig breeds. The full length cDNA of the porcine PRKAA2 (2,145 bp) obtained by RACE containing 1,656 bp open reading frame of deduced 552 amino acids, had sequence identities with PRKAA2 of human (98.2%), rat (97.8%), and mouse (97.5%). These results suggested that the porcine PRKAA2 is a positional candidate gene for fat deposition trait at near telomeric region of the long arm of SSC 6.