• BMB Reports
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• v.47 no.11
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• pp.649-654
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• 2014

Neutrophils play an important role in the initiation of innate immunity against infection and injury. Although many different types of G-protein coupled receptors are functionally expressed in neutrophils, no reports have demonstrated functional expression of umami taste receptor in these cells. We observed that mouse neutrophils express the umami taste receptor T1R1/T1R3 through RNA sequencing and quantitative RT-PCR analysis. Stimulation of mouse neutrophils with L-alanine or L-serine, which are ligands for the umami taste receptor, elicited not only ERK or p38 MAPK phosphorylation but also chemotactic migration. Moreover, addition of L-alanine or L-serine markedly reduced the production of several cytokines including $TNF-{\alpha}$ induced by lipopoly-saccharide (LPS) through inhibition of $NF-{\kappa}B$ activity or STAT3 phosphorylation in neutrophils. Our findings demonstrate that neutrophils express the umami taste receptor, through which tastants stimulate neutrophils, resulting in chemotactic migration, and attenuation of LPS-induced inflammatory response.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2190-2198
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• 2017

Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade integral chicken feathers completely at $60^{\circ}C$, resulting in the formation of 3.24 mg/ml of free amino acids from 50 ml of culture containing 10 g/l chicken feathers. Strain YT06 could grow and secrete keratinase using feather as the only carbon and nitrogen sources without other supplement, but complementation of 10 g/l sucrose and 4 g/l $NaNO_3$ increased the production of the keratinolytic enzyme. The maximum protease activity obtained was 110 U/ml and for keratinase was 42 U/ml. The keratinase maintained active status over a broad pH (pH 8-11) and temperature ($60-75^{\circ}C$). It was inhibited by serine protease inhibitors and most metal ions; however, it could be stimulated by $Mn^{2+}$ and the surfactant Tween-20. A reductive agent (${\beta}$-mercaptoethanol) was observed to cleave the disulfide bond of keratin and improve the access of the enzyme to the keratinaceous substrate. Zymogram analysis showed that strain YT06 primarily secreted keratinase with a molecular mass of approximately 35 kDa. The active band was assessed by MALDI-TOF mass spectrometry and was observed to be completely identical to an alkaline serine protease from Thermoactinomyces sp. Gus2-1. Thermoactinomyces sp. strain YT06 shows great potential as a novel candidate in enzymatic processing of hard-to-degrade proteins into high-value products, such as keratinous wastes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.5
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• pp.727-733
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• 1995

This stduy was attempted to establish the basic data for evaluating chemical components in the sap from birches(Betula platyphylla Sukatschev, Betula costata Trautv, Betula davurica Pallas), bamboos(Phyllostachys pubescens, Phyllostachys bambusoides, Phyllostachys nigra), Darae(Actinidia arguta). Calcium and potassium in five kinds of mineral detected in the sap were dominant mineral, magnesium, sodium and iron in order and calcium, potassium and magnesium are abundant in the sap from bamboo more than the other sample and the contents were 242.0~422.1mg/L, 793.8~ 2504.1mg/L and 72.6~165.9mg/L, respectively. Free sugars of the sap determined were glucose, fructose and sucrose, but maltose was not detected. The contents of glucose and fructose of the sap from Betula platyphylla Sukatschev(#2) were the highest and 42.1g/L and 36.9g/L, respectively. The detectabel nucleotides and their related compounds were CMP, UMP, GMP, IMP, AMP and hypoxanthine. The total contents of composition amino acids detected from eighteen kinds of the sap were in the range of 2.4~30.4mg%. The major amino acids were taurine, glycine, lysine, alanine and threonine in the sap from birch(#1, #2), glutamic acid and lysine in the sap from Betula costata Trauty(#3) and Betula davurica Pallas(#4), lysine, valine, alanine, serine, tyrosine and glutamic acid in the sap from bamboos, and glutamic acid, leucine, alanine in the sap from Darae.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.100-110
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• 2020

Objective: The objective of this study was to isolate proteolytic microorganisms and evaluate their effects on proteolysis in total mixed ration (TMR) silages of soybean curd residue. Methods: TMRs were formulated with soybean curd residue, alfalfa or Leymus chinensis hay, corn meal, soybean meal, a vitamin-mineral supplement, and salt in a ratio of 25.0: 40.0:30.0:4.0:0.5:0.5, respectively, on a basis of dry matter. The microbial proteinases during ensiling were characterized, the dominate strains associated with proteolysis were identified, and their enzymatic characterization were evaluated in alfalfa (A-TMR) and Leymus chinensis (L-TMR) TMR silages containing soybean curd residue. Results: Both A-TMR and L-TMR silages were well preserved, with low pH and high lactic acid concentrations. The aerobic bacteria and yeast counts in both TMR silages decreased to about 10⁵ cfu/g fresh matter (FM) and below the detection limit, respectively. The lactic acid bacteria count increased to 10⁹ cfu/g FM. The total microbial proteinases activities reached their maximums during the early ensiling stage and then reduced in both TMR silages with fermentation prolonged. Metalloproteinase was the main proteinase when the total proteinases activities reached their maximums, and when ensiling terminated, metallo and serine proteinases played equally important parts in proteolysis in both TMR silages. Strains in the genera Curtobacterium and Paenibacillus were identified as the most dominant proteolytic bacteria in A-TMR and L-TMR, respectively, and both their proteinases were mainly with metalloproteinase characteristics. In the latter ensiling phase, Enterococcus faecium strains became the major sources of proteolytic enzymes in both TMR silages. Their proteinases were mainly of metallo and serine proteinases classes in this experiment. Conclusion: Proteolytic aerobic bacteria were substituted by proteolytic lactic acid bacteria during ensiling, and the microbial serine and metallo proteinases in these strains played leading roles in proteolysis in TMR silages.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.26-32
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• 2002

For commercial production of Korean fermented anchovy sauce through rapid fermentation, a bacterial strain which showed the high protease activity was isolated from a commercially fermented anchovy sauce. The isolate was Bacillus amyloliquefaciens, and named as B. amyloliquefaciens HTP-8. The incubation temperature, initial pH, and cultivation time for optimal production of protease by B. amyloliquefaciens HTP-8 were $30^{\circ}C$, 7.0, and 3 days, respectively. In jar fermenter, B. amyloliquefaciens HTP-8 showed higher protease activity when grown at pH 7.0. The protease was partially purified by 80% ammonium sulfate precipitation and CM-Sephadex C-50 ion exchange chromatography. The partially purified enzyme had specific activity of 103.3 units/mg, yield of 0.4%, and purification fold of 43.0. The optimal pH and temperature for the protease activity were 10.0 and $50^{\circ}C$, respectively. The protease was relatively stable at the pH range of 7.0~12.0 and at the temperatures below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{＋}$ /, $Ba^{2＋}$ and selectively inhibited by PMSF, suggesting that it is a serine protease.

• Development and Reproduction
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• v.7 no.2
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• pp.113-118
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• 2003

Follicular fluid(FF) of mammalian Graafian follicles contains various kinds of proteins and proteinases that are believed to play important roles during follicular growth oocyte maturation and ovulation of mature oocytes. Previous studies of human FF(hFF) demonstrated the presence of many serine/threonine proteinases and matrix metalloproteinases such as gelatinases, however, little is known about the caseinases. Present study was aimed to examine the presence and the property of caseinolytic enzyme in hFF. Using casein zymographic method, it was found that hFF, human adult serum and cord serum exhibited one intense 80 kDa and another weak 78 kDa bands having caseinolytic activity. When inhibitors were added to the zymographic substrate buffer, caseinolytic activity of both 80 kDa and 78 kDa proteins were inhibited by othylenediarnine tetraacetic acid(EDTA) or soybean trypsin inhibitor(SBTI), but not by E-64, phenylmethylsulfonyl fluoride(PMSF) or 1,10-phenanthroline. Thus both enzymes appear to belong to a family of trypsin-like enzyme. Addition of EDTA to the zymographic substrate buffer almost abolished the caseinolytic activity of both enzymes. However, further addition of a divalent metal ion such as CaC $l_2$, MgC $l_2$, MnC $l_2$ or ZnC $l_2$ to the same buffer fully restored the enzyme activity at 5 mM concentration despite the presence of EDTA. Based upon these observations, 80 kDa and 78 kDa caseinolytic enzymes are present in human follicular fluid and they appear to be trypsin-like enzymes of which caseinolytic activity needs the presence of $Ca^{++}$, aM $g^{++}$, M $n^{++}$ or Z $n^{++}$././././.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.10
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• pp.1314-1319
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• 2007

This study is to compare food components and the sensory properties of common and functional Porphyra yezoensis, Pyroligneous liquor, Salicornia herbacea L. and Salicornia herbacea L. treated with oak charcoal. The samples used in this study were provided by a local manufacturer. We analyzed their proximate composition, mineral content, heavy metal content and amino acids. We conducted the sensory evaluation before and after grilling Porphyra yezoenis. Common components are more plentiful in functional Porphyra yezoenis than in common except for carbohydrate and crude fiber. In the case of mineral content, generally functional Porphyra yezoensis contains much more mineral than common except for magnesium and zinc. Especially Salicornia herbacea L. treated with oak charcoal contains abundant iron. Also, Salicornia herbacea L. contains a lot of calcium potassium, copper and sodium. There is a little more heavy metal except for chromium, cadmium and lead in Salicomia herbacea L. treated with oak charcoal than in common that contains a lot of chromium, cadmium and lead, although a wide difference was not discovered between them. In the case of composed amino acid, there are much glutamic acid, glycine, isoleucine and phenylalanine in both Porphyra yezoensis. Especially alanine and phenylalanine are trebled in functional Porphyra yezoensis. Also, there are far more serine and lysine in Salicornia herbacea L. and Salicornia herbacea L. treated with oak charcoal than in common Porphyra yezoensis and Pyroligneous liquor. In the case of the sensory properties, there isn't a big difference before grilling Porphyra yezoensis; however, after grilling, most functional Porphyra yezoenis had good sensory properties results, especially Salicornia herbacea L. were better than Pyroligneous liquor.

• Bulletin of the Korean Chemical Society
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• v.32 no.9
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• pp.3367-3371
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• 2011

An easy and convenient synthetic route to (S)-2-hydroxy-2'-(3-phenyluryl-benzyl)-1,1'-binaphthyl-3-carboxaldehyde (1), capable of recognizing tryptophan by fluorescence has been developed. The binol carboxaldehyde 1 exhibited a high selectivity to L-tryptophan over other examined L-${\alpha}$-amino acids such as alanine, phenylalanine, glutamine, arginine, lysine, serine, threonine, aspartat, valine, histidine and cysteine, with a fluorescence "turn-on" signal. In addition, 1 displayed chiral discrimination with good enantioselectivity toward L-tryptophan over D-tryptophan through different fluorescence enhancement factors.

• Journal of Sericultural and Entomological Science
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• v.22 no.1
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• pp.26-45
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• 1980

Silk protein is synthesized in the silkgland of silkworm, Bombyx mori L. It is evident that silk productivity is one of the high heritable characters from the genetical aspects. It is also changed with the environmental circumstances. With this regard, this paper dealt with the varietal patterns of silkgland development and the factors concerning to the silk productivity of silkgland of silkworm by the synthesis of nucleic acids, profiles of amino acids and histological basis, using the eight parent silkworm varieties and their F$_1$ hybrids. 1. The weight of silkgland per larva increased proportionally in the F$_1$ hybrids which were crossed between longer silk yielding varieties. The silk content to the weight of the silkgland was higher in the longer silk yielding varieties than that in the shorter silk yielding varieties. 2. It was observed that the morphological changes of nuclei took place in the posterior silkgland cells with the larval development of the 5th instar. In varietal aspect, Jam 107 and Jam 108, longer silk yielding varieties, showed more branches in nuclei than the $N_2$ and $C_{60}$ which were shorter yielding ones. 3. It was observed that there was a high correlation between RNA content per unit weight of silkgland on the 6th day stage of 5th instar and silk productivity both in the parents and their F$_1$ hybrids. 4. RNA and DNA synthesis brought about thirty percent increase in the posterior silkgland of the longer silk yielding varieties during the 2nd day to the 4th day stages of the 5th instar, when compared with those in the posterior silkgland of the shorter silk yielding varieties. 5. RNA/DNA ratio in the posterior silkgland on the 2nd day and 4th day stages of the 5th instar was more increased in the longer silk yielding varieties than the shorter silk yielding varieties. 6. It was shown that DNA content for the longer silk yielding varieties came to be 374$\mu\textrm{g}$ per larva in the posterior division of silkgland on the 4th day stage of 5th instar, whereas it was 199$\mu\textrm{g}$ per larva for the shorter silk yielding varieties. 7 There was 34.8％ Alanine, 22.8％ Glycine, 9.1％ Serine and 7.3％ Tyrosine in the posterior division of silkgland as major amino acids. It is noticed that there was a little differences between the amino acids composition of posterior silkgland and silk fibroin. 8. There was some differences in the amino acids composition of posterior silkgland between pure lines and their hybrids. Glycine, Serine and acidic amino acids, essential to silk formation, seemed to be increased in the F$_1$ hybrids, whereas other amino acids such as Valine, Iso-leucine, Leucine, Lysine. Phenylalanine, Histidine and Arginine were reduced. 9. The content of Glycine, Alanine and Serine in the posterior division of silkgland was elevated in the longer silk yielding varieties than the others. Consequently. these three amino acids in the posterior silkgland seemed to be related to the silk yielding ability in the longer silk yielding varieties.s.

• Molecules and Cells
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• v.19 no.2
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• pp.289-293
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• 2005

BAF53 is an actin-related protein that shuttles between nucleus and cytoplasm. In the nucleus, it constitutes an integral component of many chromatin-modifying complexes such as the SWI/SNF, TIP60, TRRAP, and TIP48/49 complexes. BAF53 is essential for growth, but its function remains elusive. BAF53 homologues from yeast to humans have a conserved N-terminal motif, MS_(G/A)(G/A)__(V/L)YGG, which is unique to these proteins. Previously we showed that over-expression of an N-terminal deletion mutant of BAF53 ($BAF53_-{\Delta}N$) reduced the viability of HEK293 and HeLa cells. When we replaced the serine 2 and tyrosine 6 of this N-terminal motif with alanine, over-expression of the alanine-replaced BAF53 strongly impaired the growth of HEK293 cells whereas replacement with aspartate/glutamate had no effect. The alanine-replaced BAF53 mutants also stimulated p53-dependent transcription, in which the SWI/SNF and TRRAP complexes are involved. Our results demonstrate that serine 2 and tyrosine 6 play important roles in BAF53 activity.