• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.179-188
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• 2017

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

• Journal of the East Asian Society of Dietary Life
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• v.12 no.1
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• pp.15-22
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• 2002

This study was carried out to investigate antimutagenic and cytotoxic effects of Korean traditional Mackjang added with sea tangle powder. The content percentage of most of minerals among general composition increased by adding sea tangle powder. By the Ames test, the antimutagenic effect of ethanol extract of Korean traditional Mackjang added with 5% sea tangle powder was strongest exceeding the control, 10%, and 15% sea tangle additions. The ethanol extract (400$\mu$g/plate) of Mackjang added with 5% sea tang1e powder in the S. typhimurium TA 100 strain showed inhibition rate of 95.0% against the mutagenesis induced by MNNG. The inhibition rate of ethanol extract (400$\mu$g/plate) of Mackjang added with 5% sea tang1e powder in the S. typhimurium TA98 and TA100 strains was 81.4% and 88.8%, respectively, against the mutagenesis induced by 4NQO. Under the same conditions, the suppression against B($\alpha$)P and Trp-P-1 in the TA98 and TA100 strains were 85.3% and 91.0%, and 96.5% and 96.5%, respectively. For the anticancer effects, an investigation was done on the cytotoxicity of Mackjang added with 5% sea tangle powder on the cell lines with human lung carcinoma (A549), human hepatocellular carcinoma (HepG2), and human gastric carcinoma (KATOIII). The cytotoxicities were inhibited with increasing the extract concentration. The treatment of 1.0 mg/mL Mackjang added with 5% sea tang1e powder showed relatively strong cytotoxicity of 61.2%, 61.8% and 66.8% against A549, KATOIII, and HepG2, respectively.

• Food Science and Preservation
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• v.9 no.1
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• pp.1-7
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• 2002

This study was carried out to investigate antimutagenic and anticancer effects of Korean traditional Kochujang added with sea tangle (Laminuia longissima). Most of the mineral content in Kochujang added with sea tangle was increased when compared to traditional Kochujang. In the Ames test, the inhibition rate of ethanol extract (200 $\mu\textrm{g}$/plate) of Kochujang added with 5% sea tangle in the S. typimurium TA100 strain showed 87.2% against the mutagenesis induced by MNNG. And also, the inhibition rate of ethanol extract (200 $\mu\textrm{g}$/plate) of Kochujang added with 5% sea tangle in the S. typhimurium TA98 and TA100 strains was 62.9% and 71.6%, respectively, against the mutagenesis induced by 4NQO. The suppression under the same condition against B($\alpha$)P and Trp-P-1 in the TA98 and TA100 strains showed 70.2% and 80.8%, and 62.9% and 73.1%, respectively. In the anticancer effects, the treatment of 1.0 mg/mL Kochujang added with 5% sea tangle showed strong cytotoxicity of 78.6% and 79.4% against KATOⅢ and HepG2, respectively.

• Applied Microscopy
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• v.41 no.3
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• pp.169-177
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• 2011

To evaluate the whitening effect of Camellia sinensis water extract (CSWE), CSWE was treated to melan-a cells. Total polyphenol contents and flavonoid contents of CSWE were 102 mg/g and 87 mg/g, respectively. The electron-donating ability of CSWE revealed a dose-dependent response, showing the excellent ability of 82% at 800 ${\mu}g$/mL, and which was higher than the arbutin (48%). The CSWE significantly (p<0.001) suppressed the melanin synthesis and the development of melanocyte dendrites was inhibited in a dose-dependent manner. The CSWE significantly (p<0.001) inhibited both intra-cellular and cell-extracted tyrosinase activities. And inhibitory efficacies of CSWE on both melanin synthesis and tyrosinase activity were significantly (p<0.001) higher than the arbutin. The tyrosinase protein expression was not influenced by arbutin treatment. However, CSWE treatment significantly (p<0.001) reduced it. Both arbutin and CSWE treatment did not influence on mRNA expressions of tyrosinase, tyrosinase-related protein-1 and tyrosinase related protein-2.

• Korean Journal of Microbiology
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• v.24 no.4
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• pp.357-363
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• 1986

Synthesis of threonine and methionine in yeast, Saccharomyces cerevisiae shares a common pathway from aspartate via homoserine. HOM6 gene encodes homoserine dehydrogenase (HSDH) which catalyzes the inter-conversion of beta-aspartate semialdehyde and homoserine. The level of HSDH is under methionine specific control. A recombinant plasmid (pEK1: 13.3kb), containing HOM6 gene, has been isolated and cloned into E. coli by complenemtary transformation of a homoserine auxotrophic yeast strain M-20-20D (hom6, trp1, ura3) to a prototrophic M20-20D/pEK1, using a library of yeast genomic DNA fragments in a yeast centromeric plasmid, YCp50(8.0kb). Isolation of HOM6has been primarily confirmed by retransformation of the original yeast strain M20-20D, using the recombinant plasmid DNA which was extracted from M20-20D/pEK1 and subsequently amplified in E. coli. Eleven cleavage sites in the insery (5.3kb) have been localized through fragment analysis for 8 restriction endonucleases; Bgl II(2 site), Bgl II(1), Cla I(3), Eco RI(1), Hind III(2), Kpn I (1), Pvu II(1) and Xho I(1).

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.2
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• pp.87-92
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• 2007

To develop the skin whitening agent, we investigated the effects of Acanthopeltis japonica, a rhodophyta on the coast of Jeju island, on melanogenesis. Dried A. japonica was refluxed with 70 % aqueous ethanol and the extract was evaporated to dryness. To validate the activity as a depigmenting agent, various in vitro tests, polyphenol contents, and free radical scavenging activity were performed. In addition, cellular tyrosinase activity and protein expression of p-ERT, tyrosinase, TRP-1, and TRP-2 were measured in B16/F10 murine melanoma cells. A. japonica had low polyphenol contents and low free radicals scavenging activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. A. japonica suppressed cellular tyrosinase activity up to 86.9 % at $100{\mu}g/mL$ with inhibition or tyrosinase and TRP-1 expression in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-treated B16/F10 melanoma cells. Our results suggest that inhibitory effects of A. japonica on melanogenesis are due to inhibiting the pathways involving ${\alpha}$-MSH-induced ERK activation. Therefore, A. japonica nay be useful as a skin whitening agent associated with the suppressive effect of melanotrophin-induced signaling pathway to inhibit melanin synthesis.

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2577-2583
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• 2011

We have designed a 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) with high bacterial cell selectivity. CAMA-P2 is an ${\alpha}$-helical antimicrobial peptide designed from a CAMA hybrid peptide and substitution of Gly-Ile-Gly hinge sequence of CAMA to Pro influences the flexibility at central part of CAMA. Based on structure-activity relationships of CAMA peptides, to investigate the effects of the total positive charges on antimicrobial activity of CAMA-P2, the $Ser^{14}{\rightarrow}$Lys analogue (CAMA-syn1) was synthesized. The role of tryptophan at C-terminal ${\alpha}$-helix on its antimicrobial activity as well as synergistic activity was also investigated using $Ser^{14}{\rightarrow}$Lys/$Phe^{18}{\rightarrow}$Trp analogue (CAMA-syn2). Also, we designed CAMA-syn3 by substitution of $Lys^{16}$ located opposite side of substituted $Lys^{14}$ of CAMA-syn1 with Leu residue, resulting in increase of hydrophobicity and amphipathicity of the peptide. All of CAMA-syn analogues showed good antimicrobial activities similar to those of CAMA and CAMA-P2. The CAMA-syn1 and CAMA-syn2 showed low hemolytic activity and cytotoxicity against human keratinocyte Haca-T cells while CAMA-syn3 showed hemolytic activity and cytotoxicity at its MIC value. We then investigated their abilities to act synergistically in combination with the antimicrobial flavonoids and synthetic compounds screened in our laboratory. The results showed that all peptides exhibited synergistic effects with dihydrobinetin, while only CAMA-syn2 exhibited synergistic effects with YKAs3001 against both S. aureus and MRSA, suggesting that Trp residue at C-terminus of CAMA-syn2 may facilitate the polar antibiotic flavonoids and synthetic compounds to permeabilize the membrane. This study will be useful for the development of new antibiotic peptides with potent antimicrobial and synergistic activity but without cytotoxicity.

• Journal of the East Asian Society of Dietary Life
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• v.11 no.1
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• pp.19-25
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• 2001

This study was performed to examine the antimutagenic and cytotoxic effects of potato vinegar and commercial vinegars(cider, brown rice, persimmon vinegars) on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, all vinegars did notexhibit any mutagenicity , but showed substantial inhibitory effects against N- methyl - N -nitro - N- nitrosog -uanidine(MNNG) , 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indol(Trp-P-1)and benzo( $\alpha$ )pyrene(B( $\alpha$ )P). The number of revertants per plate decreased significantly when these vinegars(80 ug/plate) were added to the assay system using TA100 strain. Especially, potato vinegar(80 ug/plate) showed high inhibition rate of 69.9% against mutagenicity of B( $\alpha$ )P on TA100 strain. In the cytotoxicity assay, these vinegars also showed prominent cytotoxic activity against human cancer cell lines. Potato vinegar(10 ug/well) showed the strongest cytotoxic effect against HT1080 (fibrosacoma cell) andK562 ( myelogenous leukemia) at the same concentration when compared with other vinegars.

• Toxicological Research
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• v.26 no.3
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• pp.171-175
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• 2010

The human genome contains approximately 13 orphan cytochrome P450 (P450, CYP) genes, of which the apparent function or substrate has not been identified. However, they seem to possess their own biological relevance in some tissues or developmental stages. Here, we characterized the heterologously expressed CYP2W1, an orphan P450 enzyme. The recombinant CYP2W1 protein containing a $6{\times}$(His)-tag at Nterminus has been expressed in Escherichia coli and purified. Expression level of CYP2W1 holoenzyme was around 500 nmol P450 holoenzyme per liter culture medium. The reduced CO difference spectrum of CYP2W1 showed a maximum absorption at 449 nm. CYP2W1 indicated the significant induction to bioactivate Trp-P-1, MeIQ, and IQ in E. coli DJ701 tester strain. However, the bioactivation of B[$\alpha$]P, and NNK by CYP2W1 was relatively low. The model structure of CYP2W1 suggested the characteristic P450 folds with the lengths and orientations of the individual secondary elements. The F-G loop is situated on the distal side of heme to accommodate the flexibility of active site of CYP2W1. These studies can provide useful information for the finding of its biological roles and structure-function relationships of an orphan CYP2W1 enzyme.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.1065-1070
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• 1999

This study was performed to determine the antimutagenic and cytotoxic effect of Aster scaber root ethanol extract on Salmonella typhymurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. Cancer cell lines include chronic myelogenous leukemia(K562), human gastric carcinoma(KATOIII), human hepatocellular carcinoma(Hep3B) and human breast adenocarcinoma(MCF-7). Futher fractionations with hexane, chloroform, ethyl acetate, butanol and water from ethanol extract of Aster scaber root were performed to obtain effective fraction. Ethanol extract and ethyl acetate fraction showed 79% and 82% inhibitory effect on the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) against TA100, while 48% and 60% inhibition was observed on the mutagenesis induced by 4-nitroquinoline-l-oxide(4NQO) against TA98. In the meanwhile, ethyl acetate fraction showed 78% and 85% inhibitory effect on the mutagenesis induced by benzo(${\alpha}$)pyrene[B(${\alpha}$)P] against TA98 and TA100, respectively, while 83% inhibition was observed on the mutagenesis induced by 3-amino-l,4-dimethyl-5H-pyrido(4,3-b) indole(Trp-P-1) against TA98. Ethyl acetate fraction (0.125 mg/ml) showed the strongest cytotoxic effect against K562, KATOIII, Hep3B and MCF-7 at the same concentration compared to those of other fractions. Ethanol extract and water fraction showed the least inhibitory effect.