• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• BMB Reports
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• v.29 no.3
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• pp.183-191
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• 1996

DNA fragments, homologous to the dTDP-D-glucose 4,6-dehydratase gene, obtained from the genomic DNA of Streptomyces antibioticus $T\ddot{u}99$, a producer of the unusual macrolide antibiotic chlorothricin, were cloned and sequenced. This dehydratase gene was designated as oxil. The coding region of the oxil gene is composed of 987 bp, and analysis of the DNA sequence data reveals sequences for the gene products of 329 amino acids (molecular weight of 36,037). The deduced amino acids are 59% identical to the StrE, dTDP-D-glucose 4,6-dehydratase from the streptomycin pathway. The oxil's function was examined by expressing it in E. coli using the T7 RNA polymerase/promoter system (pRSET) to produce an active fusion protein including a his tag. This enzyme shows specificity of substrate, specific only to dTDP-D-glucose.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.383-386
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• 2022

A gene encoding thermostable cellulase B (TmCelB) was isolated from Thermotoga maritima. The open reading frame (ORF) of TmCelB gene was 825bp long which predicted to encode 274 amino acid residues with a molecular weight of 31,732 Da. The 17 amino acid residues from N-terminal of the TmCelB was known as signal peptides. To analyze the enzymatic activity and biochemical properties, the ORF of TmCelB gene excluding a putative signal sequence encoding 17 amino acids were introduced into the E. coli expression vector, pRSET-B, and overexpressed in E. coli BL21. The optimum temperature of recombinant TmCelB was around 95 ℃, and the optimum pH of recombinant TmCelB was around pH 4.5. The recombinant TmCelB was stable at temperature below 100 ℃.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.777-782
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• 2004

The arylsulfatase gene (astA, 984 bp ORF) from the P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 (6.4 kb) was introduced into E. coli BL21(DE3), the transformant on the LB plate containing IPTG showed a hydrolyzing activity for 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. The highest arylsulfatase activity (2.1 unit/ml) was obtained at 10 mM IPTG. Most arylsulfatase activity was found in the cell lysate, whereas no significant activity was detected in the culture supernatant. The molecular weight of the recombinant enzyme was estimated to be 33.1 kDa by SDS-PAGE. After the reaction of agar with arylsulfatase for 12 h at $40^{\circ}C$, the gel strength of the agar increased by 2-fold, and 73% of the sulfate in the agar had been removed. This result suggests that arylsulfatase expressed in E. coli could be useful in the production of electrophoretic grade agarose.

• Journal of Life Science
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• v.5 no.4
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• pp.162-169
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• 1995

The glpD genes encoding gly-3-p dehydrogenase is essential for the aerobic growth of E. coli on glycerol or gly-3-p. The glpE gene, the function of which is unknownm is transcribed divergently with respect to glpD gene. Expression of the adjacent but divergently transcribed glpD the glpE genes is positively regulated by the cAMP-CRP complex. In this study, for a precise investigation of the functional elements in the regulatory region for transcription activation by cAMP-CRP, deletion mutation have been introducted into the regulatory region. The effect of the deletion mutant on transcriptional regulation was tested in vivo by $\beta$-galctosidase activity. Deletion mutants in the regulatory region of glpD demonstrated that the presence of the CRP-binding site resulted in an sixfold increase in promoter activity. And also deletion mutants of glpE gene demonstrated that the presence of the CRP-binding site resulted in an eightfold increase in promoter activity. Insertion of 22 bp oligomer in the deletion mutants has shown that the CRP binding site is need for maximal expression of glpD and glpE genes. glpD and glpE gene, cAMP-CRP complex, deletion mutant, transcriptional regulation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1095-1101
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• 2003

This study was carried out to investigate the effect of Gastrodiae Rhizoma (G. Rhizoma) on blood pressure-lowering in spontaneously hypertensive rats (SHR) fed high-fat diet supplemented with 10% (w/w) of lard during the experimental period of 8 weeks. Forty of male SHR weighing approximately 100 g were randomly divided into eight groups; A: negative control (lard 10%), B: positive control (lard 10% ＋ basal diet ＋ 5 brix water extract), C: lard 10% ＋ 1% G. Rhizoma powder, D: lard 10%＋5% G. Rhizoma powder, E: lard 10%＋2 brix 50% ethanol extract, F: lard 10%＋10 brix 50% ethanol extract, G: lard 10%＋2 brix water extract, H: lard 10% ＋ 10 brix water extract. A gain in weight did not differ significantly among dietary groups, but a little higher in control groups than in G. Rhizoma dietary groups. Except for spleen, weights of liver, kidney and testis are significantly different among dietary groups. Serum total cholesterol concentration was markedly higher in control groups than in G. Rhizoma groups (p<0.05), however, there was no significant difference in serum triglyceride. Except for negative control (A) and group D, serum HDL concentration was significantly higher in G. Rhizoma groups (p<0.05). On the other hand, serum LDL concentration was significantly higher in two control groups (A, B) and markedly lower in E and G groups of hot water extract of G. Rhizoma (p<0.05). Reference systolic blood pressure (BP) showed average 185.7$\pm$5.8 mmHg for 4 weeks after feeding high-fat diet, and the pressure was measured on every 7 days intervals after feeding of G. Rhizoma diet. Comparing with reference BP before feeding of G. Rhizoma diet, the groups of 50% ethanol (E, F) and water (G) extracts on BP level after 28 days were shown to be reduced at 16.8, 20.2 and 11.7 mmHg, respectively. When the pressure (187 mmHg) of group A was considered as 100%, the reduction rate of BP in group F was 11% (20.5 mmHg). These results indicated that the groups treated with ethanol extracts of G. Rhizoma showed to have lower blood pressure level compacred to the groups treated with whole powder or water extracts of G.Rhizoma in SHR fed with high-fat diet.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.111-117
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• 1995

In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.4
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• pp.561-566
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• 2014

The complete coding sequence of Wild Argali ISG15 cDNA was generated by rapid amplification of cDNA ends. The ISG15 cDNA was 642 bp with an open reading frame of 474 bp, which encoded a 17.47 kDa protein composed of 157 amino acids. Its amino acid sequence shared 97.9%, 80.8%, 91.4%, 94.3%, 78.3% identity with those of ISG15cDNA from Ovis aries (accession no. NM001009735.1), Capra hircus (accession no. HQ329186.1), Bos taurus (accession no. BC102318.1), Bubalus bubalis (accession no. HM543269.1), and Sus scrofa (accession no. EU647216.1), respectively. The entire coding sequence was inserted into the pET-28a vector and expressed in E. coli. The recombinant protein corresponded to the expected molecular mass of 25 kDa as judged by SDS-PAGE, and it was detected in the bacterial inclusion bodies. The expressed protein could be purified by $Ni^{2+}$ chelate affinity chromatography and the results from the lymphocyte proliferation test showed that the product could stimulate lymphocyte proliferation very well (p<0.05), which further confirmed its biological activity.

• ETRI Journal
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• v.46 no.3
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• pp.485-500
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• 2024

A hardware architecture is presented to decode (N, K) polar codes based on a low-density parity-check code-like decoding method. By applying suitable pruning techniques to the dense graph of the polar code, the decoder architectures are optimized using fewer check nodes (CN) and variable nodes (VN). Pipelining is introduced in the CN and VN architectures, reducing the critical path delay. Latency is reduced further by a fully parallelized, single-stage architecture compared with the log N stages in the conventional belief propagation (BP) decoder. The designed decoder for short-to-intermediate code lengths was implemented using the Virtex-7 field-programmable gate array (FPGA). It achieved a throughput of 2.44 Gbps, which is four times and 1.4 times higher than those of the fast-simplified successive cancellation and combinational decoders, respectively. The proposed decoder for the (1024, 512) polar code yielded a negligible bit error rate of 10^-4 at 2.7 E_b/N_o (dB). It converged faster than the BP decoding scheme on a dense parity-check matrix. Moreover, the proposed decoder is also implemented using the Xilinx ultra-scale FPGA and verified with the fifth generation new radio physical downlink control channel specification. The superior error-correcting performance and better hardware efficiency makes our decoder a suitable alternative to the successive cancellation list decoders used in 5G wireless communication.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.293-297
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• 2012

Of the 169 strains of microorganisms stored in Polar and Alpine Microbial Collection of Korea Polar Research Institute, 27 strains were selected for their chitinase activity on ZoBell plates supplemented with 0.4% colloidal chitin. Among them, PAMC 21693 strain have shown the highest chitinolytic enzyme activity toward pNP-$(GlcNAc)_1$ at low temperature and the highest growth rate at $4^{\circ}C$. We cloned a full-length chitinase gene of 2,857 bp which contains an open reading frame of 2,169 bp encoding 872-amino acid polypeptide. Recombinant chitinase protein was expressed in E. coli and its molecular weight was confirmed 96 kDa. In this paper, we suggest the potential use of cold-active chitinase from polar microorganisms in the field of biotechnology.