• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.921-928
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• 2002

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

• Journal of Microbiology and Biotechnology
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• v.21 no.5
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• pp.483-493
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• 2011

An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

• Journal of Microbiology and Biotechnology
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• v.21 no.10
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• pp.1021-1025
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• 2011

A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and $60^{\circ}C$, respectively. The purified chitosanase was continuously thermostable at $40^{\circ}C$. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.

• Korean Journal of Poultry Science
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• v.39 no.4
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• pp.311-315
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• 2012

The increasing demand for Korean native chicken meat indicates that the discovery of haplotypes is very important from both economic and conservation points of view. In this study, mtDNA D-loop sequences from two crossbred Korean native chicken populations of 138 individuals were investigated. Twenty six nucleotide substitutions were identified from sequence analysis and were classified into 12 haplotypes. The haplotype H_8 represents 73.47% of Woorimatdag (chicken population) sequences, which were identified in all five Woorimatdag chicken populations investigated. The H_7 haplotype (Dhap1) for D population covers 45% sequences, which indicate maternal inheritance from black Korean native chicken. On the other hand, Chap3 and Chap4 for C population are specific haplotypes, as H_5 and H_2, respectively. Based on the network profiles, six SNPs (C199T, A239G, G242A, A291G, T330C and C391A) of the D-loop region are effective markers for discrimination between Woorimatdag and Hanhyup chicken populations. Also, the phylogenetic analyses of Woorimatdag and Hanhyup chicken populations were used to identify the genetic relationships among the haplotypes. The results presented here can be used for developing molecular markers to discriminate between two commercial Korean native chickens.

• Animal cells and systems
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• v.6 no.1
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• pp.45-51
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• 2002

We compared the complete mitochondrial cytochrome b gene sequences of Korean and European cobitids to provide independent evidence for assessment of systematic and biogeographic relationships of species in the genus Cobitis. The data suggested monophyly of the genus Cobitis and the inclusion of Korean Cobitis species within the group having one lamina circularis, a primitive condition. Also, all the phylogenetic analyses using maximum parsimony, maximum likelihood, and neighbor joining methods showed a monophyletic relationship among Cobitis. The basal position of the Caspian C. cf. sibirica reported here reflects the eastern Asiatic origin cf. the European Cobitis and establishes C. cf. sibirica as an independent lineage. The Korean C. pacifica diverged next to C. cf. sibirica in basal group from the genus Cobitis. This result is in agreement with the hypothesized Asiatic origin of some European freshwater fish lineages. The phylogenetic relationships in this study showed a close affinity between C. zanadreai and C. sinensis. Two new species, C. tetralineata and C. pacifica in Korea also are closely related to monophyletic group clustering the type species of the Acanestrinia subgenus (C. elongata) with all the endemic Italian species (C. bilineata and C. zanandreai). This may suggest that the affinity between the Korean and Danubian-Italian imply genetic convergence or genetic plesiomorphic state between allopatric species that are separated for the Miocene. The mtDNA-based phylogeny for the species of the genus Cobitis from Kores and Europe permits phylogenetic assessment of the morphological transitions of Iamina circularis.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.171-177
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• 2013

The dairy industry has consistently grown via the expansion of dairy-based food categories. Dairy product consumption is stable since the nutrient composition in dairy products is ideal for human health. However, dairy products are highly susceptible to food-borne pathogens. Controlling the safety of dairy products is thus important when considering the nutrient-rich matrix of this food category. Currently, immunoassays or molecular biology techniques have been used to evaluate the safety of dairy products in Korea. These methods are based on the detection of proteins and thus have low reproducibility and sensitivity. Recent techniques to detect food-borne pathogens have focused on genetic analyses. Rapid detection methods for food-borne pathogens in milk and dairy products using polymerase chain reaction (PCR) techniques, such as conventional PCR, real-time PCR, repetitive sequence-based (rep)-PCR, PCR-denaturing gradient gel electrophoresis (DGGE), and digital PCR, are reviewed in this article. The aim of this review was to contribute knowledge of the relationship between microflora and the quality of dairy products. This study will also assist in the immediate monitoring of food-borne pathogens in milk and dairy products when an outbreak related to this food category occurs.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.139-146
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• 2006

The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

• The Journal of Korean Society of Virology
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• v.29 no.2
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• pp.65-74
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• 1999

Reovirus was found to inhabit both the respiratory and the enteric tract of human and animals. The genome of reovirus comprises 10 segments of double-stranded RNA, total size 24 kbp. Nine strains of reovirus were isolated from human and field mice in Korea. Aseptically collected sera from human and lung tissues from field mice were used for virus isolation. For serotype determination, hemagglutination inhibition test was used, and three strains were confirmed to type 2 and six strains to type 3. To determine the genomic diversity and molecular phylogeny of reoviruses isolated in Korea, part of S4 genomic segment of reovirus was enzymatically amplified and directly sequenced. In nucleotide level, Apo98-35 strain showed 15.4%, 19.3%, and 14.4% differences compared to type 1 (T1L, Lang), type 2 (T2J), and type 3 reference strains, respectively. In amino acid level, Apo98-35 strain showed 10.5%, 13.7%, and 9.5% differences compared to type 1, type 2, and type 3 reference strains, respectively. Using the maximum parsimony method based on 285 bp spaning region of the S4 genomic segment, phylogenetic analysis indicated that Apo98-35 from Korea formed different phylogenetic branch. Our data obtained by sequence and phylogenetic analyses of reoviruses are consistent with the distinct geographically dependent evolution of reoviruses in Korea.

• Fisheries and Aquatic Sciences
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• v.11 no.2
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• pp.88-97
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• 2008

Lipoproteins are complexes of lipids and specific apolipoproteins that are involved in lipid transport and redistribution among various tissues. In this study, we isolated full-length apolipoprotein cDNA sequences encoding apolipoprotein A-I (apoA-I), apoE, apoC-II, and apo-14 kDa in barred knifejaw, Oplegnathus fasciatus. In addition, we reconstructed phylogenetic trees and investigated mRNA tissue distributions. Alignment analyses of amino acid sequences revealed that secondary structures of the polypeptides apoA-I, apoE, and apoC-II in barred knifejaw are well conserved with their teleostean and mammalian counterparts in terms of characteristic tandem repetitive units forming amphipathic ${\alpha}$-helices. Both the sequence alignment data and cleavage sites of apo-14 kDa indicated a clear differentiation between Percomorpha and Cypriniformes. Meanwhile, the phylogenetic trees of apolipoprotein sub-families suggested that the common ancestor prior to the split of the Actinopterygii (ray-finned fishes) and Sarcopterygii (tetrapods) would have possessed the primordial protein-encoding genes. Tissue distribution of each apolipoprotein transcript determined by semi-quantitative RTPCR showed that barred knifejaw apoA-I transcripts were more or less ubiquitously expressed in the liver, intestines, brain, muscle, spleen, and kidney. The most striking difference from previous observations on barred knifejaw was the ubiquitous expression of apoE across all somatic tissues. Barred knifejaw apoC-II showed tissue-specific expression in the liver and intestines, while the liver and brain were the major sites of apo-14kDa mRNA synthesis.

• CELLMED
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• v.5 no.3
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• pp.15.1-15.12
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• 2015

Comparative studies have established that the North-Eastern (NE) region of India which is a part of the Eastern Himalayan region is affluent in both traditional knowledge based phytomedicine and biodiversity. About 1953 ethno-medicinal plants are detailed from the NE region of India out of which 1400 species are employed both as food and ethnopharmacological resources. Nearly 70% of species diversity has been reported from the two Indian biodiversity hotspots-The Western Ghats and the Eastern Himalayas and these hotspots are protected by tribal communities and their ancient traditional knowledge system. Paris polyphylla Smith belongs to the family Melanthiaceae and is a traditional medicinal herb which is known to cure some major ailments such as different types of Cancer, Alzheimer's disease, abnormal uterine bleeding, leishmaniasis etc. The major phytoconstituents are dioscin, polyphyllin D, and balanitin 7. Phylogeny of Paris was inferred from nuclear ITS and plastid psbA-trnH and trnL-trnF DNA sequence data. Results indicated that Paris is monophyletic in all analyses. Rhizoma Paridis, which is the dried rhizome of Paris polyphylla is mainly used in Traditional Chinese Medicine and its mode of action is known for only a few cancer cell lines. The current review determines to sketch an extensive picture of the potency, diversity, distribution and efficacy of Paris polyphylla from the Eastern Himalayan region and the future validation of its phytotherapeutical and molecular attributes by recognizing the Intellectual Property Rights of the Traditional Knowledge holders.