• BMB Reports
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• v.29 no.6
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• pp.519-524
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• 1996

Peptidylprolyl cis-trans isomerase (PPlase) catalyzes the cis-trans isomerization of prolyl peptide and facilitates the folding of cellular proteins and peptides. PPlase consists of two distinct immunophilins, each specifically binding to the immunosupressive drug cyclosporin A (CsA) or FK506, respectively. A PPlase was isolated and partially purified from porcine spleen. The molecular weight of porcine spleen PPlase was determined to be ~14,000 on the basis of SDS-PAGE. The purified enzyme was strongly inhibited by FK506, but not by CsA. The inhibition constant and the true concentration of enzyme preparations were determined by active site titration using the tight binding inhibitor FK506: $K_{i}=18.7$ nM and $E_{t}=172$ nM. The equilibrium ratio of conformer. [cis]/[trans], of prolyl peptide substrates (N-Suc-Ala-Xaa-Pro-Phe-p-NA) in anhydrous trifluoroethanol/LiCl solvent system varied from 0.24 to 0.85 depending on the nature of Xaa. Overall. in this solvent-salt system, the populations of the cis conformer of substrates in equilibrium are higher than in an aqueous solution so that the substantial error caused by high background absorption can be reduced. The reactivities of porcine spleen PPlase are shown to be highly sensitive to changes in the structure of substrates. Thus, $k_{cat}/K_m$ value for the most reactive substrate (Xaa Leu) is $4.007+10^{6}M^{1}s^{1}$ and, is 2,636 fold higher than that for the least reactive peptide substrate tested, Xaa=Glu.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.22 no.4
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• pp.49-55
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• 2021

The purpose of this study was to compare the quality characteristics of each brown rice glutinous rice red pepper paste prepared. In this study, brown rice, glutinous rice, and red pepper paste were prepared with various enzyme preparations. A 20-day study of the physicochemical and microbiological properties of each enzyme through the results process showed significant differences. In particular, brown rice red pepper paste manufactured using cozies purchased from Sunchang was functionally superior to rice cozies and bean cozies, with probiotics of 7.1-7.5×107 CFU/mL, lactobacillus of 1.9-2.2×103 CFU/mL, 48.67-52.30%, and 58.57-63.90%. When the results were combined, the composition of microorganisms and enzymes of fermented soybean paste purchased from Sunchang was not confirmed, but brown rice chili paste with nutritional and functional properties could be prepared compared to A.oryzae and A. kawachii.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.164-172
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• 2001

The angiotensin-I converting enzyme (ACE) inhibitors from laver hydrolysate was isolated. Among the 13 kinds of proteases, Maxazyme NNP was most effective for preparing the high ACE inhibitory compound. In extraction conditions of ACE inhibitory peptide from laver hydrolysate, ACE inhibitory activity of hydrolysate treated with diethylether for decolorization and that of $70\%$ ethanol soluble fraction among the different ethanol concentrations were higher than other preparations. Low molecular fraction less than 3,000 dalton of layer hydrolysate separated by ultrafiltration had the highest ACE inhibitory activity, for further separation of ACE inhibitory peptide from laver hydrolysate, gel filtration chromatography (Sephadex G-25), reverse-phase HPLC (ODS & Vydac C-18) and gel permeation chromatography (Superdex Peptide HR) were performed. The molecular mass of the ACE inhibitory peptide fractions of gel permeation chromatography determined by electrospray-mass spectrometer were 413.48 (S1O2V2V1P),346.86 (S1O2V2V2P) and 320.32 (S2O6V3V1P) dalton and their amino acid sequence were Val-Gln-Gly-Asn, Thr-Glu-Thr and Phe-Arg, respectively.

• Korean Journal of Food Science and Technology
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• v.24 no.6
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• pp.519-523
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• 1992

The patterns on the proteolysis of mussel protein using a commercial enzyme preparation were investigated. The best one among six commercial enzyme preparations for the manufacture of mussel extract was Corolase PP, based on the degree of hydrolysis (DH). When the raw mussel paste, without water addition, was adjusted to pH 6.5, added 0.1% (w/w dry basis) of Corolase PP. and reacted at $50^{\circ}C$ for four hours, it reached the maximum value of DH (79%). The precooking of raw mussel decreased the efficiency of extraction and hydrolysis of the protein, due to the inactivation of the autolytic enzymes contained in the mussel. During the course of proteolysis, major free amino acids such as glycine, alanine, glutamic acid and lysine, representing a characteristic brothy taste of mussel were replaced with free hydrophobic amino acids including valine, methionine, isoleucine, and leucine. The electrophoretic pattern and HPLC-GPC pattern of mussel protein hydrolysates during the hydrolysis were observed and also discussed.

• Korean Journal of Agricultural Science
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• v.21 no.2
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• pp.142-147
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• 1994

Characteristics of TNP-cellulose which prepared from carboxymethyl cellulose powder, CM32, as substrate for cellulase activity assay were investigated. Enzymatic hydrolysis of TNP-cellulose occured on the cellulose moiety but not on amide bonds, following Michaelis-Menten kinetics. Three cellulase preparations from Trichoderma viride, Aspergillus niger, and Cellulomonas sp. were tested for their pH and temperature dependences and compared with the method determining the increase in reducing power. The enzyme activity was found to have the same temperature range in both methods, however the pH range was broadened in the case of using TNP-cellulose as substrate. The colorimetric method for cellulase assay using TNP-cellulose as substrate was compared with the other methods: one based on determination of the increase in reducing power; and the other based on determining the decrease in viscosity of Na-CM-cellulose solution. The activities measured by the colorimetric method showed a linear correlation with the enzyme concentration of certain range in all three enzymes tested, and the activity values were proportional to those obtained from the other methods. Depending on the enzyme, however, the activity values from this method were not always in proportion to those from the viscometric method. suggesting that this method was not specific for determination of the endo-type cellulase.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.1
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• pp.7-13
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• 2001

Enzymatic hydrolysis of different forms of silk fibroin (soluble, gel and insoluble forms) by industrial and commercial enzyme preparations to obtain aqueous and powdered silk fibroin in relatively mild conditions was investigated. A mono-enzymatic hydrolysate systems were tested for hydrolysis of water-soluble form of fibroin as most productive form of protein substrate. Insoluble forms of substrate usually were hydrolyzed less effective. In some cases from soluble fibroin substrate gel was formed during hydrolysis process. This hindered intermixing and decreased rates of hydrolysis. Insoluble sediments were formed in enzymatic hydrolysates in other cases. These sediments and also sediment after chemical hydrolysis were purified and tested on amino acids content for comparison. Sediments formation in these conditions are considered as pure tyrosine isolation method. Obtained hydrolysates were characterized by gel-chromatography analysis and other standard biochemical methods. Possibility of application of enzymatic hydrolysis for preparation of silk fibroin hydrolysates is discussed.

• Journal of Nutrition and Health
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• v.17 no.1
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• pp.60-67
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• 1984

Inhibiting action of sulfur dioxide $(SO_{2})$ on enzymatic discoloration was investigated with crude enzyme preparations from homogenized tissues or sliced disks of raw potato tubers. $SO_{2}$ appeared to inhibit the formation of dopachrome in a competitive manner. At insufficient concentrations of $SO_{2}, the formation of dopachrome was reinitiated as time elapsed. The present results suggested that $SO_{2}$ would form an additional compounds with certain intermediate during the course of enzymatic browning and delay the enzymatic discoloration. To inhibit the production of dopachrome in sliced disks of raw potato tubers, much higher concentrations of $SO_{2}$ were required than homogenized tissues.

• Applied Microscopy
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• v.13 no.1
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• pp.49-61
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• 1983

Protoplast releasing mechanisms from Trichoderma koningii, fine structures of the released protoplsts, and their regeneration mode were studied by scanning and transmission electron microscopy. Two types of protoplast releasing mechanisms were observed. In one mechanism, cytoplasm emerged through a cell wall pore developed by cell lytic enzymes and formed a spherical protoplast. In the other mechanism, as the cell wall became progressively thinner, the inner cytoplasm partially rounded to form nonspherical bodies which became spherical protoplasts after being released into the enzyme solution. But, these two types of protoplast releasing mechanisms did not seem to be. mutually exclusive but could occur on the same mycelium simultaneously. And it appeared that cytoplasm which did not become a protoplast by the first mechanism could from a protoplast by the second mechanism. The preparations contained two types of protoplasts, released from different sites of the mycelia. Those released from younger mycelia had dense cytoplasm and small vesicles. Those released from the older mycelia had less dense cytoplasm and larger vacuoles. In the case of regeneration, before producing normal mycelia, most of the protoplasts assumed aberrant tube and yeast-like-forms. Normal mycelia were produced at the end of the yeastlike-forms and sometimes in the middle of the aberrant tube.

• Journal of the Korean Society of Tobacco Science
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• v.25 no.1
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• pp.70-79
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• 2003

$\beta$ -Glucosidase-catalysed synthesis of glucosides with aromatic alcohols and monoterpene alcohols as accepters and cellobiose as a donor in the presence of various commercial $\beta$ -glucosidases were described. $\beta$ -Glucosidases from Aspergillus niger spp,. Trichoderma spp., Penicillium sup. and bitter almond have been shown to catalyze synthesis of $\beta$ -glucosides of benzyl alcohol, 2-hydroxybenzyl alcohol, 4-hydroxybenzyl alcohol, 2-phenylethyl alcohol, geraniol and citronellol in the presence of cellobiose as sugar donor. Among enzyme preparations tested, each $\beta$ -glucosides prepared from Aspergillus niger were isolated in the pure state by Diaion HP-20 and silica gel column chromatography. The products were identified as $\beta$ -glucosyl products of benzyl alcohol, 2-hydroxyhenzyl alcohol, 4-hydroxybenzyl alcohol, 2-phenyl ethyl alcohol, geraniol and citronellol by spectrometry (UV, IR, $^1$H-NMR, $^{13}$ C-NMR) and enzymatic hydrolysis with $\beta$ - glucosidase. Monoterpene alcohols with a sterically hindered hydroxyl group, such as linalool, $\ell$-menthol and $\alpha$-terpineol were not used as acceptors in transglycosylation reaction.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.318-324
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• 2017

With the development of next-generation sequencing (NGS), a cutting-edge technology, genotype-by-sequencing (GBS) became available at a low cost per sample. GBS makes it possible to customize the process of library preparation to obtain high-quality single nucleotide polymorphisms (SNPs) in the most efficient way. However, a GBS library is hard to construct due to fine-tuning of concentration of each reagent and set-up. The major reason for this is the presence of undigested genomic DNA (gDNA) owing to the efficiency of different restriction enzymes for different species with unknown reasons. Therefore, this proof-concept study is to demonstrate the unpredictable patterns of enzyme digestion from various plants in order to make the reader aware of the caution needed when choosing restriction enzymes for their GBS library preparations. Indeed, no pattern was found for the digestibility of gDNA samples and restriction enzymes in the current study. We suggest that more data should be accumulated on this matter to help researchers who want to apply GBS technologies in a variety of genetic approaches.