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전통누룩 진균류를 이용한 입국의 제조 및 입국곰팡이의 동정
김재호 ( Jae Ho Kim ),권영희 ( Young Hee Kwon ),이애란 ( Ae Ran Lee ),김혜련 ( Hye Ryun Kim ),안병학 ( Byung Hak Ahn ) 한국균학회 2012 韓國菌學會誌 Vol.40 No.4
다양한 향미를 가진 막걸리의 개발을 위해 전통누룩으로부터 분리한 곰팡이로 입국을 제조한 후 품질특성을 분석하여 입국의 규격에 적합하며 이취가 없고 관능이 우수한 9균주를 입국 제조용 우수균주로 최종 선발하였다. 선발된 균주는 Aspergillus oryzae(C1-5-2-2, C20-7-3, CN1.3.1-4, CN16.19.1-1, N152-1, N220-1), Mycocladus corymbiferus (N162-2), Rhizopus oryzae(N20), Lichtheimia corymbifera (N21)로 동정되었으며, 제조한 입국의 산도는 5.0~6.8, 당화력은 128~241sp이었다. Various koji were prepared by fungi isolated from traditional nuruk and their quality characteristics were investigated. Acidity and saccharification power of their koji were ranged in 5.0~6.8 and 128sp~241sp. Nine fungi which were showed good quality and sensory evaluation were identified by analysis of their nucleotide sequences with PCR-amplified 18S rDNA internal transcribed spacer-1(ITS-1) and ITS-4 genes. Among them, six strains were identified as Aspergillus oryzae and the other strains were identified as Mycocladus corymbiferus, Rhizopus oryzae, Lichtheimia corymbifera.
Purification and preliminary analysis of the lysophospholipase from Cutibacterium acnes
Ae-Ran Kwon 한국구조생물학회 2020 Biodesign Vol.8 No.1
Acne is one of the most common dermatological conditions, but the detailed pathology is unclear and current treatment regimens are plagued by side effects. Cutibacterium acnes is known as a major acne associated bacterium that derives energy from lipase mediated sebum lipid degradation. C. acnes is commensal, but free fatty acids digested by C. acnes lipases from triglyceride and lysophosphatidyl choline induce marked inflammation. Up to date, it remains to be elucidated the lipase activity mechanism at the molecular level. In this study, we report the purification and crystallization of the fulllength lysophospholipase from C. acnes. The crystal diffracted X-rays to a 1.7 Å resolution, revealing that the crystals belong to space group P21212, with cell dimension parameters of a = 94.5, b = 129.7, and c = 55.5 Å. Two protomers are present in the crystallographic asymmetric unit, with a calculated crystal volume per protein weight (VM) of 2.50 Å3Da-1 and a solvent content of 50.79%. This structure will contribute towards revealing a detailed mechanism of acne development.
Kwon, Ae-Ran,Kim, Ji-Hun,Park, Sung Jean,Lee, Ki-Young,Min, Yu-Hong,Im, Hookang,Lee, Ingyun,Lee, Kyu-Yeon,Lee, Bong-Jin Oxford University Press 2012 Nucleic acids research Vol.40 No.9
<P>VapD-like virulence-associated proteins have been found in many organisms, but little is known about this protein family including the 3D structure of these proteins. Recently, a relationship between the Cas2 family of ribonucleases associated with the CRISPR system of microbial immunity and VapD was suggested. Here, we show for the first time the structure of a member of the VapD family and present a relationship of VapD with Cas2 family and toxin–antitoxin (TA) systems. The crystal structure of HP0315 from <I>Helicobacter pylori</I> was solved at a resolution of 2.8 Å. The structure of HP0315, which has a modified ferredoxin-like fold, is very similar to that of the Cas2 family. Like Cas2 proteins, HP0315 shows endoribonuclease activity. HP0315-cleaved mRNA, mainly before A and G nucleotides preferentially, which means that HP0315 has purine-specific endoribonuclease activity. Mutagenesis studies of HP0315 revealed that D7, L13, S43 and D76 residues are important for RNase activity, in contrast, to the Cas2 family. HP0315 is arranged as an operon with HP0316, which was found to be an antitoxin-related protein. However, HP0315 is not a component of the TA system. Thus, HP0315 may be an evolutionary intermediate which does not belong to either the Cas2 family or TA system.</P>
ermK Leader Peptide : Amino Acid Sequence Critical for Induction by Erythromycin
Kwon, Ae-Ran,Min, Yu-Hong,Yoon, Eun-Jeong,Kim, Jung-A,Shim, Mi-Ja,Choi, Eung-Chil The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.12
The ermK gene from Bacillus lichenformis encodes an inducible rRNA methylase that confers resistance to the macrolide-lincosamide-streptogramin B antibiotics. The ermK mRNA leader sequence has a total length of 357 nucleotides and encodes a 14-amino acid leader peptide together with its ribosome binding site. The secondary structure of ermK leader mRNA and a leader peptide sequence have been reported as the elements that control expression. In this study, the contribution of specific leader peptide amino acid residues to induction of ermK was studied using the PCR-based megaprimer mutation method. ermK methylases with altered leader peptide codons were translationally fused to E. coli ${\beta}-galactosidase$ reporter gene. The deletion of the codons for Thr-2 through Ser-4 reduced inducibility by erythromycin, whereas that for Thr-2 and His-3 was not. The replacement of the individual codons for Ser-4, Met-5 and Arg-6 with termination codon led to loss of inducibility, but stop mutation of codon Phe-9 restored inducibility by erythromycin. Collectively, these findings suggest that the codons for residue 4, 5 and 6 comprise the critical region for induction. The stop mutation at Leu-7 expressed constitutively ermK gene. Thus, ribosome stalling at codon 7 appears to be important for ermK induction.
Role of Disulfide Bond of Arylsulfate Sulfotransferase in the Catalytic Activity
Kwon, Ae-Ran,Choi, Eung-Chil The Pharmaceutical Society of Korea 2005 Archives of Pharmacal Research Vol.28 No.5
Bacterial arylsulfate sulfotransferase (ASST) catalyzes the transfer of sulfate group from a phenyl sulfate ester to a phenolic acceptor. The promoter region and the transcripti on start sites of Enterobacter amnigenus astA have been determined by primer extension analysis. Northern blot analysis resolved two mRNA species with lengths of 3.3 and 2.0 kb, which correspond to the distances between the transcriptional initiation sites and the two inverted repeat sequences (IRSs). By length, the 3.3 kb RNA could comprise the three-gene (astA with dsbA and dsbB) operon. ASST has three highly conserved cysteine residues. Reducing and non-reducing SDS-PAGE and activity staining showed that disulfide bond is needed for the activity of the enzyme. To identify the cysteine residues responsible for the disulfide bond formation, a series of Cys to Ser mutants has been constructed and the enzymatic activity was measured. Based on the results, we assumed that the first cysteine (Cys349) might be involved in disulfide bond mainly with the second cysteine (Cys445) and result in active conformation.
Kwon, Ae-Ran,Yoon, Eun-Jeong,Yoon, Jong-Min,Kim, Hyun-Jee,Shim, Mi-Ja,Choi, Eung-Chil 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.7
The antibacterial activities of clindamycin, synercid, telithromycin, linezolid and mupirocin were evaluated against erythromycin-resistant Gram-positive coccal clinical isolates collected in Korean hospitals. In Staphylococcus aureus, synercid, linezolid and mupirocin were the most active agents. Against coagulase-negative staphylococci (CNS), synercid, linezolid and mupirocin were also active. Telithromycin and synercid resistance was common against enterococci, only linezolid and mupirocin were active. The reason of low activity of telithromycin against staphylococci and enterococci is because most of the isolates were constitutively resistant to erythromycin. Synercid, telithromycin, linezolid and mupirocin were active against streptococci.
( Ae Ran Lee ),( Jeong A Park ),( Young Eun Kim ),( Hyung Joo Kwon ),( Young Hee Lee ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.14
Stromal cell-derived factor-1 (SDF-1/CXCL12) enhances the survival of hematopoietic stem cells, progenitor cells, and a human progenitor cell line MO7e, especially in synergy with other cytokines. Previously, we reported that c-Fos was synergistically induced by combined stimulation with SDF-1/CXCL12 plus other cytokines. Here, we aimed to evaluate that induced c-Fos expression is required for the survival enhancement in MO7e cells. First by using RNase protection assay, we confirmed that synergistic induction of c-Fos was unique among several stress-related genes. Since c-Fos is known as one of the components of transcription factor AP-1, we performed electrophoretic mobility shift assay. The DNA binding activity of AP-1 was also synergistically enhanced by combined stimulation, and c-Fos was identified in the DNA bound complex. To further evaluate the significance of c-Fos expression for survival of MO7e cells, we employed cell permeable c-Fos dominant negative mutant form (A-Fos-MTS) and protein transduction procedure. Inhibition of c-Fos by treatment with A-Fos-MTS significantly reduced survival of MO7e cells in the presence of SDF-1/CXCL12 plus low concentration of GM-CSF. Therefore, we suggest that induction of c-Fos and enhanced AP-1 activity contribute to survival effect of cytokines in MO7e cells.
In vitro and in vivo activities of DW-224a, a novel fluoroquinolone antibiotic agent
Kwon, Ae-Ran,Min, Yu-Hong,Ryu, Jei-Man,Choi, Dong-Rack,Shim, Mi-Ja,Choi, Eung-Chil Oxford University Press 2006 The Journal of antimicrobial chemotherapy Vol.58 No.3
<P><I>Objectives</I>: The objective of the present study was to assess the <I>in vitro</I> and <I>in vivo</I> activities of DW-224a in order to eventually use it as an antibiotic.</P><P><I>Methods</I>: DW-224a was compared with DW286, ciprofloxacin and trovafloxacin. MICs of DW-224a, DW286, ciprofloxacin and trovafloxacin were determined against several groups of clinical isolates. In addition, intraperitoneal infection was induced with various organisms in mice. Test compounds were administered once orally to mice immediately after infection. The 50% protective dose (PD<SUB>50</SUB>) was calculated from the survival rates on day 7 after infection.</P><P><I>Results</I>: Against Gram-positive bacteria, the <I>in vitro</I> activity of DW-224a was stronger than those of ciprofloxacin and trovafloxacin, but slightly weaker than that of DW286. Against Gram-negative bacteria, the activity of DW-224a was similar to those of trovafloxacin and DW286, but weaker than that of ciprofloxacin. In experimental systemic infections in mice with various organisms, like DW286, DW-224a demonstrated potent activity against Gram-positive bacteria and somewhat less activity against Gram-negative bacteria.</P><P><I>Conclusions</I>: DW-224a has a broad spectrum of antimicrobial activity, which is especially potent against Gram-positive bacteria.</P>