• Molecules and Cells
• /
• 제28권6호
• /
• pp.575-581
• /
• 2009

Ornithine decarboxylase (ODC) is a rate-limiting enzyme in the biosynthesis of polyamines, which are essential for cell growth, differentiation, and proliferation. This report presents the characterization of an ODC-encoding cDNA (SlitODC) isolated from a moth species, the tobacco cutworm, Spodoptera litura (Lepidoptera); its expression in a polyamine-deficient strain of yeast, S. cerevisiae; and the recovery in polyamine levels and proliferation rate with the introduction of the insect enzyme. SlitODC encodes 448 amino acid residues, 4 amino acids longer than B. mori ODC that has 71% identity, and has a longer C-terminus, consistent with B. mori ODC, than the reported dipteran enzymes. The null mutant yeast strain in the ODC gene, SPE1, showed remarkably depleted polyamine levels; in putrescine, spermidine, and spermine, the levels were > 7, > 1, and > 4%, respectively, of the levels in the wild-type strain. This consequently caused a significant arrest in cell proliferation of > 4% of the wild-type strain in polyamine-free media. The transformed strain, with the substituted SlitODC for the deleted endogenous ODC, grew and proliferated rapidly at even a higher rate than the wild-type strain. Furthermore, its polyamine content was significantly higher than even that in the wild-type strain as well as the spe1-null mutant, particularly with a very continuously enhanced putrescine level, reflecting no inhibition mechanism operating in the putrescine synthesis step by any corresponding insect ODC antizymes to SlitODC in this yeast system.

• 폴리머
• /
• 제30권5호
• /
• pp.373-379
• /
• 2006

플로로글루시놀과 중합가능한 피리딘 유도체의 수소결합을 이용해 디아세틸렌과 아크릴로일 그룹을 함유 한 새로운 중합가능한 원반형 액정들을 제조하였고 제조된 액정들의 광중합 거동을 조사하였다. 합성된 원반형 액정 복합체들은 메소겐의 방향족 고리의 개수에 따라 원반형 컬럼상과 장방형 컬럼상을 형성하였다. 원반형 액정복합체들의 광중합은 액정상에서 자외선을 조사하여 수행하였다. 자외선 조사 후 디아세틸렌과 아크릴로일 그룹이 선택적으로 중합되었으며, 짧은 공액구조의 디아세틸렌 올리고머를 가지는 가교 고분자들이 1,4-반응에 의해 형성됨을 적외선 분광법과 자외선-가시광선 분광분석을 통해 확인하였다. X-선 회절 실험 결과, 페닐피리딘을 함유한 원반형 액정 복합체의 컬럼상 질서는 광중합 후에도 유지되었고, 바이페닐 단위를 지닌 액정 복합체의 장방형 컬럼상 질서는 라멜라 질서로 변화됨을 확인하였다.

• Bulletin of the Korean Chemical Society
• /
• 제31권11호
• /
• pp.3359-3365
• /
• 2010

The incorporation of microwave irradiation with the prevalent "click chemistry" is currently of considerable synthetic interest. We describe here the introduction of such laboratorial shortcut into carbohydrate-based drug discovery, resulting in the rapid formation of a series of triazole-linked salicylic $\beta$-D-O-glycosides with biological activities. All "clicked" products were achieved in excellent yields ($\approx$ 90%) within only a quarter. In addition, based on the structural characteristics of the afforded glycomimetics, their inhibitory activities were evaluated toward protein tyrosine phosphatases 1B (PTP1B) and a panel of homologous protein tyrosine phosphatases (PTPs). Docking simulation was also conducted to plausibly propose binding modes of this glycosyl salicylate series with the enzymatic target.

• 원예과학기술지
• /
• 제34권1호
• /
• pp.32-45
• /
• 2016

Fruit bagging has been widely practiced in peach cultivation to produce high quality and unblemished fruit. Moreover, fruit bagging has been utilized to study the effect of shading on the quality of fruit. We conducted a proteomic analysis on peach fruit to elucidate the biochemical and physiological events that characterize the effect of bagging treatment. Comparative analysis of 2D electrophoresis (2-DE) gels showed that relative protein levels differed significantly at 125 DAFB (days after full bloom), as well as at 133 DAFB in fruit that had been bagged until 125 DAFB, followed by exposure to sunlight. Most of the proteins with altered expression were identified by MALDI TOF/TOF. Twenty-one proteins with differential expression among the groups were identified at 125 DAFB, while thirty proteins with differential expression among the groups were identified at 133 DAFB. The analysis revealed that expression of proteins involved in photosynthesis, stress responses, and biochemical processes influencing metabolism were altered during bagging treatment, suggesting that regulation of the synthesis of carbohydrates, amino acids, and proteins influenced fruit size, solid/acid ratio, and peel color. This work provides the first characterization of proteomic changes in peach in response to fruit bagging treatment. Identifying and tracking protein changes may allow us to better understand the mechanisms underlying the effects of bagging treatment.

• Journal of Microbiology and Biotechnology
• /
• 제30권5호
• /
• pp.785-792
• /
• 2020

ʟ-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce ʟ-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the ʟ-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50℃, 7, and 5 mM MnCl₂, respectively. Additionally, ATP was found to be an important factor for producing high concentration of ʟ-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher ʟ-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM ʟ-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of ʟ-theanine.

• 유기물자원화
• /
• 제31권1호
• /
• pp.57-68
• /
• 2023

본 연구에서는 커피 폐기물 기반의 질소가 포함된 다공성 탄소 섬유 형태로 제조하여 고에너지 EDLC용 탄소 소재로 활용하고자 하였다. 커피 폐기물은 분쇄과정을 거쳐 폴리비닐피롤리돈과 용매인 다이메틸폼아마이드에 혼합한 후 전기방사를 통해 커피 폐기물 기반의 섬유 형태(Bare-CWNF)의 물질로 만들었으며, 질소 분위기의 900℃에서 탄화를 진행하여 커피 폐기물 기반의 질소가 포함된 다공성 섬유 형태(Carbonized-CWNF)의 물질을 제조하였다. Carbonized-CWNF는 Bare-CWNF와 같이 섬유 형태를 유지하였으며 질소 함량 역시 유지되는 것을 확인하였다. 커피 폐기물의 탄화 탄소(Carbonized-CW)및 폴리아크릴로나이트릴 기반의 탄소섬유(Carbonized-PNF)를 Carbonized-CWNF와 -1.0-0.0V의 전압 범위에서 전기화학적 성능을 비교한 결과, Carbonized-CWNF가 가장 높은 비정전용량(123.8F g^-1 @ 1A g^-1)을 확보할 수 있었다. 이를 통해 커피 폐기물 기반의 질소가 함유된 다공성 탄소 섬유가 고에너지 EDLC(Electric double layer capacitor)용 전극으로 우수한 성능을 나타내는 것을 확인하였다. 최종적으로, 환경 오염의 원인이 되는 식물성 바이오매스 중 커피 폐기물을 활용하여 친환경성을 확보하였고, 식물성 바이오매스와 같은 폐기물을 슈퍼커패시터와 같은 고성능 에너지 저장 매체로의 탈바꿈 할 수 있는 가능성을 제시하였다.

• 한국수산과학회지
• /
• 제28권2호
• /
• pp.209-218
• /
• 1995

본 연구에서는 유전자 복제기작을 생화학적, 분자생물학적 방법을 사용하여 bacteriphage T7을 대상으로 연구하였다. Bacteriophage T7의 유전자 복제, 재조합, 수선시 필수 단백질로 작용하는 gene 2.5 단백질의 생체내 기능에 대한 유전학적 연구와 단백질을 분리 정제하여 복제 단백질들과의 상호작용에 대한 연구를 수행하였다. 연구결과 gene 2.5 단백질은 DNA복제시 필수 구성단백질로 작용하며, 복제과정에서 유전자 복제에 관여하는 핵심 단백질들인 DNA polymerase, helicase/primase와 직접 단백질-단백질 상호 협동 작용을 하는 r것을 증명하였다. 특히 gene 2.5 단백질의 C-terminal domain이 절편된 변이체의 경우 복제 단백질들과 상호작용이 결여되었다. 따라서 C-terminal domain이 gene 2.5 단백질의 기능에 필수적으로 관여함을 입증하였다.

• Journal of Periodontal and Implant Science
• /
• 제24권1호
• /
• pp.144-154
• /
• 1994

Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

• 생명과학회지
• /
• 제31권9호
• /
• pp.862-872
• /
• 2021

최근 생물의학 분야에서의 광범위한 응용 가능성에 의하여 식물이나 미생물을 이용한 은(Ag), 금(Au), 백금(Pt), 세륨(Ce), 아연(Zn), 구리(Cu) 등의 금속 나노물질 합성에 관한 연구가 주목받고 있다. 식물은 플라보노이드, 알칼로이드, 사포닌, 스테로이드 탄닌과 각종 영양 성분과 같은 생리 활성 물질을 풍부하게 가지고 있으며, 유사하게 미생물들도 단백질과 같은 생리활성 대사산물이나 색소, 항생제 및 산과 같은 가치가 있는 화학물질을 분비한다. 최근 보고된 바에 의하면, 나노입자의 생체 합성은 무해한 방법일 뿐만 아니라 항균, 항진균, 세포 증식 억제 및 항플라스모디아 활성과 같은 생물의학 분야로서의 적용에 주요한 후보로 여겨진다. 나노입자의 이러한 생리 활성은 농도에 의존적이며, 나노입자의 모양과 크기에도 따라 달라질 수 있다. 미생물과 식물은 나노입자의 친환경적합성에 사용되는 대사산물이나 화학물질 등의 훌륭한 공급원으로서 생물 의학 분야에서 유용하게 사용된다. 미생물 또는 식물 원료를 사용하여 합성된 나노입자는 화학적인 방법으로 합성된 나노입자보다 더 낮은 독성을 나타낸다. 본 리뷰논문에서는 미생물이나 식물과 같은 생물학적 재료를 이용한 나노입자의 합성과, 합성에 사용되는 다양한 기술의 특성 및 나노입자의 항균 분야에서의 적용에 대하여 중점적으로 서술하였다.

• Journal of Microbiology
• /
• 제45권6호
• /
• pp.534-540
• /
• 2007

The Streptomyces coelicolor A3(2) genome contains six operons (rrnA to F) for ribosomal RNA synthesis. Transcription from rrnD occurs from four promoters (p1 to p4). We found that transcripts from the p1 and p3 promoters were most abundant in vivo in the early exponential phase. However, at later phases of exponential and stationary growth, transcripts from the p1 promoter decreased drastically, with the p3 and p4 transcripts constituting the major forms. Partially purified RNA polymerase supported transcription from the p3 and p4 promoters, whereas pure reconstituted RNA polymerase with core enzyme (E) and the major vegetative sigma factor ${\sigma}^{HrdB}$ ($E{\cdot}{\sigma}^{HrdB}$) did not. In order to assess any potential requirement for additional factor(s) that allow transcription from the p3 and p4 promoters, we fractionated a partially purified RNA polymerase preparation by denaturing gel filtration chromatography. We found that transcription from the p3 and p4 promoters required factor(s) of about 30-35 kDa in addition to RNAP holoenzyme ($E{\cdot}{\sigma}^{HrdB}$). Therefore, transcription from the p3 and p4 promoters, which contain a consensus -10 region but no -35 for ${\sigma}^{HrdB}$ recognition, are likely to be regulated by transcription factor(s) that modulate RNA polymerase holoenzyme activity in S. coelicolor.