• Journal of Life Science
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• v.14 no.2
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• pp.325-330
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• 2004

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (Homologue of RAD3 gene). The over-expressed HRD3 protein was estimated to be a 75 kDa in size which is in good agreement with the estimated by the nucleotide sequence of the cloned gene. Two-dimensional gel electrophoresis showed that a number of other protein spots dramatically disappeared when the HRD3 protein was overexpressed. The overexpressed RAD3 protein showed a toxicity in E. coli host, suggesting that this protein may be involved in the inhibition of protein synthesis and/or degradation of host protein. To determine which part of HRD3 gene contributes to the toxicity in E. coli, various fusion plasmids containing a partial sequence of HRD3 and lac'Z gene were constructed. These results suggest that the C-terminal domain of HRD3 protein may be important for both toxic effect in E. coli and for its role in DNA repair in S. pombe.

• Journal of Life Science
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• v.30 no.1
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• pp.106-112
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• 2020

Aminoacyl tRNA synthetase complex interacting multifunctional protein 2 (AIMP2) is a scaffolding protein required for the assembly of multi-tRNA synthetase, and it can exert pro-apoptotic activity in response to DNA damage. In the presence of DNA damage, AIMP2 binds to mouse double minute 2 homolog (MDM2) to protect p53 from MDM2 attack. TGF-β signaling results in the nuclear translocation of AIMP2, whereby AIMP2 interacts with FUSE-binding protein, and, thus, suppresses c-myc. TNF receptor-associated factor 2 (TRAF2) is an important mediator between TNF-receptors 1 and 2 which are involved in the signaling of c-Jun N-terminal kinase (JNK), nuclear factor κB (NF-κB), and p38 mitogen-activated protein kinases (MAPKs). TRAF2 is required for the activations of JNK and NF-κB via TNF-α and the mediation of anti-apoptosis signaling. AIMP2 can also enhance pro-apoptosis in the TNF-α signaling. During this signaling, AIMP2 assists the association of E3 ubiquitin ligase, the cellular inhibitor of apoptosis protein 1 (c-IAP1) which is well known and responsible for the degradation of TRAF2. The formation of a complex among AIMP2, TRAF2, and c-IAP1 results in proteasome-mediated TRAF2 degradation. AIMP2 can induce apoptosis via downregulation of TRAF2 to interact directly in TNF-α signaling. This review provides new insight into the molecular mechanism responsible for AIMP2 and TRAF2 complex formation and treatments for TNFα-associated diseases.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.1
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• pp.68-72
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• 2004

The effects of the addition of yeast on in vitro roughage degradability and methane production were investigated in order to clarify the effects of yeast on the rumen microbes and to establish methods of rumen manipulation. Three roughages (whole crop corn, rice straw and Italian ryegrass) were incubated for 3, 6, 12 and 24 h with or without dried beer yeast following the method described by Tilley and Terry. Using the same method, these roughages were incubated with or without yeast extract, albumin or purified DNA. In vitro methane production was measured with or without dried beer yeast at 12 and 24 h. The degradability of yeast was found to be 57 and 80% at 12 and 24 h, respectively. The rate of degradation of fraction b was 6.16%/h. There was a significant increase in roughage degradability at 6 h (p<0.05), 12 h (p<0.05) and 24 h (p<0.01) by dried yeast addition. The degradability of all three roughages was higher in the samples treated with yeast extract than in the no addition samples except in the case of rice straw incubated for 12 h. Nevertheless, the magnitude of increment was smaller with the addition of yeast extract than without the addition of yeast. With the addition of purified DNA, there were significant increases in roughage degradability at 6 h (p<0.01), 12 h (p<0.01) and 24 h (p<0.05); however, higher degradability values were detected in the samples to which albumin was added, particularly at 6 h. If the degradability values of the no addition samples with those of samples containing yeast, yeast extract, DNA and albumin were compared, the largest difference was found in the samples to which yeast was added, although it is worth noting that higher values were observed in the yeast extract samples than in the DNA or albumin samples, with the exception of the case of rice straw incubated for 24 h. Methane production was significantly increased at both 12 and 24 h incubation. The increment of roughage degradation and methane production brought about by the addition of dried beer yeast to the samples was thought to be due to the activation of rumen microbes. Water soluble fraction of yeast also seemed to play a role in ruminal microbe activation. The increment of degradability is thought to be partially due to the addition of crude protein or nucleic acid but it is expected that other factors play a greater role. And those factors may responsible for the different effects of individual yeast on ruminal microbes.

• Journal of Animal Science and Technology
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• v.47 no.4
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• pp.615-624
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• 2005

The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.

• Korean Journal of Microbiology
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• v.33 no.2
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• pp.149-156
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• 1997

We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.

• Journal of Life Science
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• v.12 no.3
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• pp.340-348
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• 2002

Equisetum arvense L. and Lactuca dentata Makino. var. Flaviflora Makino. of samples relatively showed anticancer effects on MCF-7 mammary gland adenocarcinoma cell. The most active plant among the samples was Capsicum annuum L. var angulosum Mill. We studied that MCF-7 cells were changing chromosomal morphology and apoptosis on these samples. Capsicum annuum L. var. angulosum Mill. of samples relatively showed good anticancer effects. The cells became vague after 2 days and then destroyed. The supernatant of the cells including medium was measured by UV absorbance. The results showed that Capsicum annuum L. var. angulosum Mill also exerted high level. We also used electrophoresis in order to observe apoptic characterization of DNA fragmentation. The cells treated with Capsicum annuum L. var. angulosum Mill showed the apoptotic characterization. The chromosome of the cells were observed on those samples. The cells treated with Capsicum annuum L. var. angulosum Mill among them were shown the fastest changes. The cells were aggregated and destroyed by treatment with some edible plants. Especially, the case of Capsicum annuum L. var. angulosum Mill, it led MCF-7 cell to apoptosis faster than others. And we can observe chromosomal changes and dispersion by PI staining. These results showed that each sample exerted anticancer effects on MCF-7 cells. Especially Capsicum annuum L. var. angulosum Miff exerted significant anticancer effects.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.198-205
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• 2011

This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.483-489
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• 2004

Pseudomonas sp. S-47 is a bacterium capable of degrading benzoate as well as 4-chlorobenzoate (4CBA). Benzoate and 4CBA are known to be degraded via a meta-cleavage pathway characterized by a series of enzymes encoded by xyl genes. The meta-cleavage pathway operon in Pseudomonas sp. S-47 encodes a set of enzymes which transform benzoate and 4CBA into TCA cycle intermediates via the meta-cleavage of (4-chloro )catechol to produce pyruvate and acetyl-CoA. In the current study, the meta-pathway gene cluster was cloned from the chromosomal DNA of S-47 strain to obtain pCS1, which included the degradation activities for 4CBA and catechol. The genetic organization of the operon was then examined by cloning the meta-pathway genes into a pBluescript SKII(+) vector. As such, the meta-pathway operon from Pseudomonas sp. S-47 was found to contain 13 genes in the order of xylXYZLTEGFlQKIH. The two regulatory genes, xylS and xylR, that control the expression of the meta-pathway operon, were located adjacently downstream of the meta-pathway operon. The xyl genes from strain S-47 exhibited a high nucleoside sequence homology to those from Pseudomonas putida mt-2, except for the xylJQK genes, which were more homologous to the corresponding three genes from P. stutzeri AN10. One open reading frame was found between the xylH and xylS genes, which may playa role of a transposase. Accordingly, the current results suggest that the xyl gene cluster in Pseudomonas sp. S-47 responsible for the complete degradation of benzoate was recombined with the corresponding genes from P. putida mt-2 and P. stutzeri AN10.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.114-121
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• 2012

The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

• Journal of the Korea Organic Resources Recycling Association
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• v.14 no.4
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• pp.141-150
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• 2006

To develop the microbial agents for the environmentally-friendly treatment and recycling of food waste, useful microorganisms, which showed higher degradation activities to various organic compounds and possessed flocculating activities, were isolated from the earthworm. One of the isolated strains, named JS-01, was further selected due to its higher flocculating activity against 0.5% Kaolin clay. JS-01 was identified as Bacillus pumilus sp. by 16s rDNA analysis. The optimal temperature and pH for the growth of JS-01 to express the flocculating activity was found to be at $37^{\circ}C$ in pH 7.0 of EPS broth medium. JS-01 also expressed good degrading activity against cellulose, which is one of the representative organic materials in food waste. We propose that JS-01 will be a good candidate for the efficient treatment of food waste and leachate due to the property to degrade cellulose and flocculating activity.