• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.905-913
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• 2014

Lj-RGD3, an RGD (Arg-Gly-Asp) toxin protein from the salivary gland of Lampetra japonica, exhibits antifungal activity against Candida albicans. Lj-RGD3 has three RGD motifs and shows homology to histidine-rich glycoprotein. We synthesised two mutant derivatives of Lj-RGD3: Lj-26, which lacks all three RGD motifs and contains no His residues; and Lj-112, which lacks only the three RGD motifs. We investigated the effects of the wild-type and mutated toxins on a gram-positive bacterium (Escherichia coli), a gram-negative bacterium (Staphylococcus aureus), and a fungus (C. albicans). rLj-RGD3 and its mutants exhibited antifungal but not antibacterial activity, as measured by a radial diffusion assay. The C. albicans inhibition zone induced by rLj-112 was larger than that induced by the other proteins, and its inhibitory effect on C. albicans was dose-dependent. In viable-count assays, the rLj-112 MIC was $7.7{\mu}M$, whereas the MIC of the positive control (ketoconazole) was $15{\mu}M$. Time-kill kinetics demonstrated that rLj-112 effectively killed C. albicans at $1{\times}$ and $2{\times}$ MIC within 12 and 6 h, respectively. Electron microscopy analysis showed that rLj-RGD3 and rLj-112 induced C. albicans lysis. Our results demonstrate a novel anticandidal activity for rLj-RGD3 and its mutant derivatives.

• Journal of Ginseng Research
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• 제36권2호
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• pp.176-189
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• 2012

This study examined the effects of Korean red ginseng (KRG) on obese women and aimed to confirm that the effects of KRG on obesity differ dependently on a gene. Fifty obese women were recruited and randomized to receive KRG (n=24) or placebo (n=26) for 8 wk. Measurements of blood pressure, height, weight, waist circumference, waist-hip ratio (WHR), total fat mass, percentage of body fat, resting metabolic rate, basal body temperature, and daily food intake (FI), blood test (serum lipid, liver and renal function), Korean version of obesity-related quality of life scale (KOQOL), and a gene examination were performed. Comparisons of subjects before and after the administration of KRG revealed significant improvements of obesity in terms of weight, body mass index (BMI), WHR, FI, and KOQOL. However, in the comparison between KRG group and placebo group, only KOQOL was significantly different. KRG displayed significant efficacy on BMI and KOQOL in the CT genotype of the G protein beta 3 gene, but not in the CC genotype, on blood sugar test in the Trp64/Arg genotype of the beta 3 adrenergic receptor gene, but not in Trp64/Trp genotype, on KOQOL in the DD genotype of the angiotensin I converting enzyme gene, but not in the ID and DD genotypes. The effects of KRG on obesity were confirmed to some extent. However, a distinct effect compared to placebo was not confirmed. KRG is more effective for improving the secondary issues of the quality of life derived from obesity rather than having direct effects on the obesity-related anthropometric assessment and blood test indices.

• Clinical and Experimental Reproductive Medicine
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• 제27권2호
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• pp.173-177
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• 2000

연구목적: 한국여성의 다낭성 난포증후군 환자에서 황체형성 호르몬 exon 2 유전자의 변이를 탐색하여 이들 변이와 질환과의 관련성 여부를 밝히고자 하였다. 연구재료 및 방법: 21명의 다낭성 난포증후군 환자를 대상으로 황체형성 호르몬 exon 2(Trp8Arg;TGG to CGG and Ile15Thr; ATC to ACC)의 변이를 탐색하였다. 혈액에서 Genomic DNA를 추출하여 PCR로 증폭한 후 RFLP 방법으로 변이형을 구분하였다. 결과: 황체형성 호르몬 exon 2의 변이형이 다낭성 난포증후군 환자에서 28.6%로 이미 조사된 바 있는 대조군의 16.7% 보다 약간 높게 나타났으나 통계적으로 유의한 차이는 없었다(p>0.05). 결론: 황체형성 호르몬 exon 2의 변이가 한국인의 다낭성 난포증후군 발병과 관련이 있는지를 밝히기 위해 더 많은 개체에 대한 연구가 요구된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권2호
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• pp.179-182
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• 2000

This study was performed to determine whether the $LH{\beta}$-subunit gene missense mutation is present in Korean infertile patients with 46,XX POF women. The variants of $LH{\beta}$ exon 2 (Trp 8Arg; TGG to CGG and Ile15Thr; ATC to ACC) were studied in forty-four 46,XX idiopathic POF and 54 nonpregnant women. The $LH{\beta}$ exon 2 variants were more frequent in POF patients (20.5%) than nonpregnant (16.7%) women (p>0.05). POF patients with the variant was slightly higher than nonpregnant women with the variant.

• Genomics & Informatics
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• 제12권4호
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• pp.268-275
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• 2014

The harshness of legionellosis differs from mild Pontiac fever to potentially fatal Legionnaire's disease. The increasing development of drug resistance against legionellosis has led to explore new novel drug targets. It has been found that phosphoglucosamine mutase, phosphomannomutase, and phosphoglyceromutase enzymes can be used as the most probable therapeutic drug targets through extensive data mining. Phosphoglucosamine mutase is involved in amino sugar and nucleotide sugar metabolism. The purpose of this study was to predict the potential target of that specific drug. For this, the 3D structure of phosphoglucosamine mutase of Legionella pneumophila (strain Paris) was determined by means of homology modeling through Phyre2 and refined by ModRefiner. Then, the designed model was evaluated with a structure validation program, for instance, PROCHECK, ERRAT, Verify3D, and QMEAN, for further structural analysis. Secondary structural features were determined through self-optimized prediction method with alignment (SOPMA) and interacting networks by STRING. Consequently, we performed molecular docking studies. The analytical result of PROCHECK showed that 95.0% of the residues are in the most favored region, 4.50% are in the additional allowed region and 0.50% are in the generously allowed region of the Ramachandran plot. Verify3D graph value indicates a score of 0.71 and 89.791, 1.11 for ERRAT and QMEAN respectively. Arg419, Thr414, Ser412, and Thr9 were found to dock the substrate for the most favorable binding of S-mercaptocysteine. However, these findings from this current study will pave the way for further extensive investigation of this enzyme in wet lab experiments and in that way assist drug design against legionellosis.

• 한국식품과학회지
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• 제17권5호
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• pp.371-376
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• 1985

내한유채를 $25^{\circ}C$에서 60시간동안 발아시켜 건물량, 뿌리신장도, 아미노산 및 지방산조성의 변화를 조사한 결과는 다음과 같다. 유채의 건물량은 발아30시간에서 비교적 급격한 감소를 나타냈고 그 이후는 완만한 감소를 나타냈으며, 우리는 15시간까지는 관찰되지 않았으나 발아30시간이후는 일정한 속도의 신장도를 나타낸다. 유채는 발아함에 따라 수분, 조회분은 큰 영향을 받지 않았으나 조지방 조단백질은 감소하였으며 조섬유는 증가하였다. 유채의 아미노산은 Glu, Lys, Asp가 많고 lie, Met가 적었으며 제 1제한 아미노산은 Ile 이었다. 유채의 아미노산은 발아함에 따라 Lys, His, Arg, Asp, Thr, Ser, Ala, Val, lie, Leu는 증가하였지만 Clu, Met Tyr, Phe은 감소하여 발아함에 따라 Met이 제 1제한아미노산이 되었다. 지방산은 oleic acid (49.3%). linoleic acid(22.0%), eicosenoic acid (10.5%), Palmitic acid(5.9%), linolenic acid(5.7%), stearic acid(3.2%), erucic acid(2.8%)가 검출되었는데. 발아에 따라 oleic acid는 감소하였으며 Palmitic acid, linolenic acid 발아중기까지 증가하다가 그 이후는 감소하였고 linolenic acid eicosenoic acid 및 erucic acid는 발아중기까지는 감소하다가 그 이후는 증가하였다.

• KSBB Journal
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• 제10권3호
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• pp.343-348
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• 1995

LeI은 스테로이드를 통울에 투여하였을 때 분비가 촉진되어 항염증성 효과를 나타내는 calcium 의 존성 phospholipid 결합 단백질이다. S. cerevisiae는 대장균과 통물세포의 장점을 모두 가지고 있으므로 동물세포 유래의 이종 단백질의 분비 생산에 많이 이용되고 있다. 본 연구에서는 GAL10 promoter­p ppL-LCI유전자 LCI terminator로 구성된 pYGLPT5 로 LCI을 S. cerevisiae SEY2102에서 발현 분비시키고 각 분획으로 나누어 LCI양을 비교한 결과 protoplast 68.6 %. periplasmic 24 %, culture supernatant 7.4%로 분포하였다. pYGLPT5로 형질전환된 S. cereviswe 2102를 유가 배양한 결과, 최종적인 LCI의 생산량은 약 $500mg/\ell$ 였다. LCI은 N 말단 부근에 $CA^{++}$ 결합부위가 있으므로 이를 이용하여 hydroxylapatite column chromatography로 정 제하 였다. 배지로 분비된 34kDa LCI을 ultrafiltration 과 hydroxylapatite column chromatography 의 두 단계로 순도 99% 이상으로 정제할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1077-1086
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• 2016

Glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone (DHA). The gldA gene encoding a putative GlyDH was cloned from Thermoanaerobacterium thermosaccharolyticum DSM 571 (TtGlyDH) and expressed in Escherichia coli. The presence of Mn²⁺ enhanced its enzymatic activity by 79.5%. Three highly conserved residues (Asp¹⁷¹, His²⁵⁴, and His²⁷¹) in TtGlyDH were associated with metal ion binding. Based on an investigation of glycerol oxidation and DHA reduction, TtGlyDH showed maximum activity towards glycerol at 60℃ and pH 8.0 and towards DHA at 60℃ and pH 6.0. DHA reduction was the dominant reaction, with a lower K_m(DHA) of 1.08 ± 0.13 mM and V_max of 0.0053 ± 0.0001 mM/s, compared with glycerol oxidation, with a K_m(glycerol) of 30.29 ± 3.42 mM and V_max of 0.042 ± 0.002 mM/s. TtGlyDH had an apparent activation energy of 312.94 kJ/mol. The recombinant TtGlyDH was thermostable, maintaining 65% of its activity after a 2-h incubation at 60℃. Molecular modeling and site-directed mutagenesis analyses demonstrated that TtGlyDH had an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg⁴³, both related to dinucleotide binding.

• Applied Biological Chemistry
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• 제5권
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• pp.39-42
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• 1964

1) 진석화(眞石花)젓은 그 수분함량(水分含量)이 생(生)굴에 비(比)하여 상당(相當)히 감소(減少)하여 농축(濃縮)된 상태(狀態)이며, glycogen의 함량(含量)도 격감(激減) 하였음에 반(反)하여 유리당(遊離糖)은 현저(顯著)하게 증가(增加)되었다. 2) Paper chromatography로 시료(試料) 즙액중(汁液中)에서 검출(檢出) 확인(確認)된 유리(遊離) 아미노산(酸)는 aspartic acid, glutamic acid, cystine, serine, glycine, lysine, threonine histidine, alanine, arginine, valine, methionine, tyrosine, phenylalanine, leucine, iso-leucine, tryptophan 등 17종(種)이 였으며, 영양상(榮養上) 필수(必須) amino acid를 전부(全部) 함유(含有)하고 있었다. 3) 비색정량법(比色定量法)에 의(依)해서 정량(定量)된 시료(試料) 즙액중(汁液中) 유리(遊離) 아미노산(酸)의 함량(含量)은 Tyr. 0.264mg/ml, Arg. 0.304mg/ml, Try. 0.046mg/ml, His. 0.460mg/ml, Met. 0.350mg/ml, Cys. 0.440mg/ml, phe. 0.115mg/ml 이었다.

• Macromolecular Research
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• 제16권6호
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• pp.517-523
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• 2008

A RGD (Arg-Gly-Asp) conjugated chitosan hydrogel was used as a cell-supporting scaffold for articular cartilage regeneration. Thermosensitive chitosan-Pluronic (CP) has potential biomedical applications on account of its biocompatibility and injectability. A RGD-conjugated CP (RGD-CP) copolymer was prepared by coupling the carboxyl group in the peptide with the residual amine group in the CP copolymer. The chemical structure of RGD-CP was characterized by $^1H$ NMR and FT IR. The concentration of conjugated RGD was quantified by amino acid analysis (AAA) and rheology of the RGD-CP hydrogel was investigated. The amount of bound RGD was $0.135{\mu}g$ per 1 mg of CP copolymer. The viscoelastic parameters of RGD-CP hydrogel showed thermo-sensitivity and suitable mechanical strength at body temperature for cell scaffolds (a> 100 kPa storage modulus). The viability of the bovine chondrocyte and the amount of synthesized glycosaminoglycans (GAGs) on the RGD-CP hydrogels were evaluated together with the alginate hydrogels as a control over a 14 day period. Both results showed that the RGD-CP hydrogel was superior to the alginate hydrogel. These results show that conjugating RGD to CP hydro gels improves cell viability and proliferation, including extra cellular matrix (ECM) expression. Therefore, RGD conjugated CP hydrogels are quite suitable for a chondrocyte culture and have potential applications to the tissue engineering of articular cartilage tissue.