• The Korean Journal of Mycology
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• v.17 no.4
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• pp.223-228
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• 1989

This study was performed to obtain higher yield rate of protein-bound polysac-charide extracted from cultured broth and fruit body of Coriolus versieolor, basidiomycetes. 16002 strain produced 0.58%, the highest yield rate among four strains, and there was no significantly different yield in other three strains. The optimal liquid media were CVT-III to obtain higher yield of protein-bound polysaccharide comparing with the various culture media. Compared with several materials for substitution of medium source, the residue extract media of Coriolus versieolor fruit-body were yield rate similar to control, and addition of the extracts of ginseng residues increased 11.4-48%. And also compared with fruit bodies that were artificially cultured on the oak longs, 16002 strain produced the highest yield similar to the result of cultured broth. Fruit body of the prematures stage was more effective to obtain high yield rate.

• Korean Journal of Environmental Agriculture
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• v.37 no.2
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• pp.97-103
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• 2018

BACKGROUND: Dissipation of acetamiprid and thiamethoxam in greenhouse grown chard samples was evaluated at 5 intervals including the pre-harvest interval after application. This study was conducted to determine the residue levels, the biological half-lives and dissipation rate of acetamiprid and thiamethoxam in chard under controlled conditions. METHODS AND RESULTS: Acetamiprid and thiamethoxam were applied in accordance with good agricultural practices for chard. Chard samples were collected at 0, 1, 2, 3, 5, 7, 10 and 14 days after application. Quantitaion was performed by HPLC-DAD system with C18 column. Limit of quantification (LOQ) of acetamiprid and thiamethoxam were both 0.02 mg/kg for chard. The recovery of acetamiprid and thiamethoxam were 77.8~107.5% and 94.3~108.6% at two concentration levels. The half-lives of pesticide dissipation in chard for two fields were 11.9 and 8.2 days for acetamiprid and 3.6 and 3.3 days for thiamethoxam respectively. The dissipation rate of acetamiprid and thiamethoxam were estimated according to the statistics method with a 95% confidence. CONCLUSION: Dissipation of acetamiprid and thiamethoxam in chard were determined under greenhouse. The concentration of acetamiprid and thiamethoxam in chards at 0 days after application were below specified by Korean MRL. Dissipation rate constant will be useful to set the pre-harvest residue limit for public health and consumer protection.

• BMB Reports
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• v.31 no.4
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• Journal of Microbiology and Biotechnology
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• v.29 no.12
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• pp.1882-1893
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• 2019

β-Glucosidases and β-xylosidases are two categories of enzymes that could cleave out non-reducing, terminal β-D-glucosyl and β-D-xylosyl residues with release of D-glucose and D-xylose, respectively. In this paper, two functional β-glucosidase Dth3 and β-xylosidase Xln-DT from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and Xln-DT were relatively stable at 75℃ and were tolerant or even stimulated by glucose and xylose. Dth3 was highly tolerant to glucose with a K_i value of approximately 3 M. Meanwhile, it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the βglucosidase Dth3 and β-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6 and C-3 sugar moieties of ASI, which established an effective and green method to produce the more pharmacologically active CAG, an exclusive telomerase activator. We measured temperature, pH and dosage of enzyme using a single-factor experiment in ASI biotransformation. After optimization, the optimal reaction conditions were as follows: 75℃, pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5% within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific, environment-friendly and cost-effective way to produce CAG.

• Journal of Pharmaceutical Investigation
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• v.37 no.4
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• pp.217-222
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• 2007

To develop cleansing tissue composed of silicone multiple emulsions which could easily remove make-up residues and confer skin protecting effect without damaging skin, we formulated various silicone multiple emulsions and evaluated the physicochemical properties including viscosity, cleansing effect, and safety effect. Also, cleansing tissue incorporated with silicone multiple (W/Si/W) emulsion was stored for 6 months, and judged its stability through microscopes under accelerated and long-term condition. Cleansing effect was evaluated by chromameter. Skin hydration effect was determined by corneometer and incorporation effect into non-woven fabric cotton was evaluated by volunteer survey. Low viscosity ranged from 400 centipoise (cP) to 1,000 cP was obtained from a stabilized W/Si/W emulsion containing more than 10% volatile silicone. Mean diameter of fresh W/Si/W emulsion was $20{\mu}m$, but after storage for 3 months at $45^{\circ}C$, the particle size of the W/Si/W emulsion increased up to $50{\mu}m$. Both W/Si/W emulsion-incorporated cleansing tissue and commercial product showed equally good cleansing effect. In addition, skin allergies such as erythema, edema, scaling itching, stinging, burning, tightness and prickling were not observed through macroscopic examination. From the transepidermal water loss results, the cleansing tissue consisting of W/Si/W emulsion showed superior hydration effect to commercial product. In conclusion, this study suggests cleansing tissue using W/Si/W emulsion could be used for an excellent efficacy compared with commercialized cleansing tissue.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.1
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• pp.1-5
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• 2002

A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2036-2045
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

• Mass Spectrometry Letters
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• v.14 no.2
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• pp.42-47
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• 2023

Carthamus tinctorius L. (known as safflower) is a valuable oil plant whose importance is increasing rapidly in the world due to its high adaptation to arid regions. The seeds of this unique plant are especially used in edible oil, soap, paint, varnish and lacquer production. Its flowers are used in vegetable dye production and medicinal purposes beside its features as a coloring and flavoring in food. After the oil is removed, the remaining pulp and plant parts are used as animal feed, and dry straw residues are used as fuel. Beside all these features, its usage as a herbal medicinal plants for various diseases has gained importance on recent years. In this study, it was designed a plant metabolomic approach which transfers all the recent data processing strategies of untargeted metabolomics in clinical applications to the present study. Q-TOF LC/MS-based analysis of the extracts (70% ethanol, hexane, and chloroform) for both seed and flowers was performed using a C18 column (Agilent Zorbax 1.8 µM, 100 × 2.1 mm). Differences were observed in seed and fruit extracts and these differences were visualized using principal component analysis (PCA) plots. The total number and intersections of the peaks in the extracts were visualized using peak count comparison graph. Based on the experimental results, the number of the detected peaks for seeds was higher than the ones for the flowers for all solvent systems to extract the samples.

• Archives of Pharmacal Research
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• v.23 no.6
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• pp.633-636
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• 2000

Translationally controlled tumor protein (TCTP), also known as IgE-dependent histamine-releasing factor, is a growth-related tumor protein. Although the primary sequence of rat TCTP does not reveal any recognizable $Ca^{2+}$ -binding motif, previous studies have demonstrated that rat TCTP consisting of 172 amino acids is a $Ca^{2+}$ -binding protein. However. the region of TCTP required for $Ca^{2+}$ interaction has not been mapped to the molecule. Here, we reported that the $Ca^{2+}$ binding region of TCTP which was mapped by using a combination of deletion constructs of rat TCTP and $^{45}Ca^{2+}$-overlay assay. was confined to amino acid residues 81-112. This binding domain did not show any peculiar loop of calcium- binding motif such as CaLB domain and EF hand motif and it seems to be constituted of random coil regions neighboring the a helix. Thus, our data confirm that TCTP is a novel family of $Ca^{2+}$ -binding protein.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.459-464
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• 1997

The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a .gamma. gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. There were three potential asparagine-linked glycosylation sites at residues 67, 68, and 315. In order to obtain UDPGTh2 protein encoded from cloned human liver UDP-glucuronosyltransferase cDNA, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. The presence of a transferase with Mr-52,000 in transfected cells cultured in the presence of $[^{35}S]$ methionine was shown by immunocomplexed products with goat antimouse transferase IgG and protein A-Sepharose and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The expressed UDPGT was a glycoprotein as indicated by electrophoretic mobility shift in Mr-3,000-4,000 when expressed in the presence of tunicamycin. The extent of glycosylation was difficult to assess, although one could assume that glycosyl structures incorporated at the level of endoplasmic reticulum were always the core oligosaccharides. Thus, it is likely that at least two moieties inserted can account for the shift of Mr-3,000-4,000. This study demonstrates the cDNA and deduced amino acid sequence of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2.