• 제목/요약/키워드: Tripeptide

검색결과 42건 처리시간 0.023초

Portage Transport of Toxophoric Agent, N-hydroxyalanine, through Oligopeptide Permease in Escherichia coli

  • Hong, Nam-Joo;Park, Yeong-Taek
    • Bulletin of the Korean Chemical Society
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    • 제14권6호
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    • pp.674-678
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    • 1993
  • Di-and tri-peptides containing DL-N-hydroxyalanine were prepared. DL-N-Hydroxyalanine was linked, via its primary amino group, to the ${\alpha}$-carbon of glycine residues in dipeptide synthon (L-alanyl(${\alpha}$-DL-N-hydroxyalanyl)DL-glycine) 5, and tripeptide synthon (L-alanyl-L-alanyl(${\alpha}$-DL-N-hydroxyalanyl) DL-glycine) 12. 5 proved to be 19 times more potent than DL-N-hydroxyalanine when tested in vitro for the ability to inhibit the growth of E coli. However, 12 gave comparable potency to DL-N-hydroxyalanine itself.

Expression of Antihypertensive Peptide, His-His-Leu, as Tandem Repeats in Escherichia coli

  • Jeong, Do-Won;Shin, Dong-Seok;Ahn, Chang-Won;Song, In-Sang;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.952-959
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    • 2007
  • His-His-Leu (HHL), a tripeptide derived from a Korean soybean paste, is an angiotensin-I-converting enzyme (ACE) inhibitor. We report here a method of producing this tripeptide efficiently by expressing tandem multimers of the codons encoding the peptide in E. coli and purifying the HHL after hydrolysis of the peptide multimers. The HHL gene, tandemly multimerized to a 40-mer, was ligated with ubiquitin as a fusion gene (UH40). UH40 was inserted into vector pET29b; the UH40 fusion protein was then produced in E. coli BL21. The recombinant UH40 protein was purified by cation-exchange chromatography with a yield of 17.3mg/l and analyzed by matrixassisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry and protein N-terminal sequencing. Leucine aminopeptidase was used to cleave a 405-Da HHL monomer from the UH40 fusion protein and the peptide was purified using reverse-phase high-performance liquid chromatography (HPLC) on a C18 HPLC column, with a final yield of 6.2mg/l. The resulting peptide was confirmed to be HHL with the aid of MALDI-TOF mass spectrometry, glutamine-TOF mass spectrometry, N-terminal sequencing, and measurement of ACE inhibiting activity. These results suggest that our production method is useful for obtaining a large quantity of recombinant HHL for functional antihypertensive peptide studies.

The penetration enhancement and the lipolystic effects of TAT-GKH, in both In vitro, Ex vivo, and In vivo.

  • Lim, J.M.;Chang, M.Y.;Park, S.G.;Kang, N.G.;Song, Y.S.;Lee, Y.H.;Yoo, Y.C.;Cho, W.G.;Han, S.G.;Kang, S.H.
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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    • pp.87-107
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    • 2003
  • It was demonstrated that Transactivating transcriptional activator(TAT) protein from HIV-1 shown to enter cells when added to the surrounding media. TAT peptide chemically attached to various proteins was able to deliver these proteins to various cell and even in tissues in mice with high levels in heart and spleen. In this study, the tripeptide GKH(Glycine-Lysine-Histidine) derived from Parathyroid hormone (PTH), which was known as lipolytic peptide, is attached to 9-poly Lysine(TAT) to be used as a cosmetic ingredient for slimming products. When Glycerol release, expressed as extracellular glycerol concentration, is lipolysis index, TAT-GKH at $10^{-5}$mo1/L induces approximately 41.5% maximal lipolytic effects in epididymal adipocytes isolated from rats, compared with basal lipolysis. Epididymal adipose tissues of male rats is assessed ex vivo by microdialysis. Probes are perfused with Ringer solution in which increasing concentrations of TAT-GKH. The perfusion of TAT-GKH induces lipolytic effect. Penetration study showed that TAT-GKH efficiently elevates 36 times higher penetration into the excised hairless mice skin than GKH. in vivo study showed that TAT-GKH had a better effect upon the relative volume of eye bag after 28 days of application on twenty(+2) healthy female volunteers. It was identified that TAT-GKH increases penetration enhancement and lipolytic effects in both in vitro, ex vivo and in vivo.

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Degumming of Antheraea yamamai silkworm cocoon

  • Shin, Bong-Seob;Jeon, Jong-Young;Kim, Jong-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제31권2호
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    • pp.127-131
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    • 2015
  • Oak silkworm, Antheraea yamamai (A. yamamai), has been used for clothing and surgical suture and considered as biomaterial due to RGD tripeptide. This paper reported the degumming conditions of A. yamamai using sodium oleate, high pressure and temperature, and sodium carbonate. Degumming ratio of A. yamamai cocoon using sodium oleate was less than 10%. High pressure and temperature treatment induced 30% weight loss of A. yamamai cocoon. The concentration, treatment temperature and time using sodium carbonate was examined and revealed the following conditions for degumming; 5% owf, 60 min at 100℃. The degummed solution was confirmed using UV and FT-IR spectrometer. Our results can be used to handle A. yamamai silkworm cocoon for further application including material processing.

Identification of a Gene Required for Gliding Motility in Myxococcus xanthus

  • Lee Cha-Yul;Chung Jin-Woo;Kim Ji-Hoon;Cho Kyung-Yun
    • Journal of Microbiology and Biotechnology
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    • 제16권5호
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    • pp.771-777
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    • 2006
  • A novel gene (agiA) required for adventurous gliding motility in Myxococcus xanthus has been identified. Null mutations in this gene caused defects in the gliding movement of isolated cells, suggesting that it belongs to one of the A-motility genes. The isolated agiA mutant cells neither glided nor produced slime trails on agar surface. However, agiA was different from other known A-motility genes in that the agiA mutant created in the $S^-$ mutant background glided in the swarm of cells, since other known A-motility mutants created in the $S^-$ mutant background do not move in the swarm of cells. The agiA mutant was also defective in fruiting body development. Sequence analysis predicted that agiA encodes a 787-amino-acid protein with eight tripeptide repeat motifs.

Synthesis of $\alpha$-L-Aspartyl-L-phenylalanine Methyl Ester from an Artificial Polypeptide

  • Choi, Soon-Yong;Kim, Hyun-Soo;Lee, Se-Yong
    • Journal of Microbiology and Biotechnology
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    • 제2권1호
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    • pp.1-6
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    • 1992
  • The aspartame, $\alpha$-L-aspartyl-L-phenylalanine methylester, is an artificial sweetener. Taking advantage of the fact that the aspartame is a derivative of dipeptide, synthesis of aspartame from the artificial polypeptide made by an artificial gene has been attempted. The artificial polypeptide (LAP32), a polymer of tripeptide (aspartyl-phenylalanyl-lysine), was purified from the E. coli cells harboring a recombinant plasmid containing the artificial gene. This polypeptide was then digested with trypsin and carboxypeptidase B to produce dipeptide (Asp-Phe). Using the esterase activity of $\alpha$-chymotrypsin, the dipeptide was directly converted into Asp-Phe methylester in a water-methanol system. When the methanol concentration in reaction mixture was 25%, 50% of dipeptide was converted to the dipeptide methylester without producing any by-products.

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숫컷 생쥐에서 타우린 투여에 의한 간내 글루타치온의 감소 (Reduction of Hepatic Glutathione by Acute Taurine Treatment in Male Mice)

  • 이선영;곽혜은;김영철
    • 약학회지
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    • 제47권4호
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    • pp.218-223
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    • 2003
  • Effect of taurine treatment on metabolism of glutathione (GSH) was studied in adult male ICR mice. An acute injection of taurine (250 mg/kg, ip) resulted in a significant decline of hepatic GSH level at t = 6 hr, but plasma GSH level was not altered. The activity of GSH-related enzyme in liver, such as GSH peroxidase, GSSG reductase, GSH S-transferases, ${\gamma}$-glutamylcysteine synthetase or ${\gamma}$-glutamyltranspeptidase, was not affected by taurine at t = 2.5 or 6 hr. Plasma cysteine and cystine levels were elevated rapidly following taurine treatment. Hepatic cysteine level was decreased by taurine, reaching a level approximately 70% of control at t = 4 and 6 hr. In conclusion, the results indicate that an acute dose of taurine decreases hepatic GSH level by reducing the availability of cysteine, an essential substrate for synthesis of this tripeptide in liver. It is also suggested that taurine may decrease the cysteine uptake by competing with this S-amino acid for a non-specific amino acid transporter.

Chromatographic Determination of Amino Acids in Nonprotein and Protein Fraction Of Undaria Pinnatifida

  • 권태완;이태영
    • Applied Biological Chemistry
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    • 제1권
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    • pp.55-61
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    • 1960
  • Ion 교환수지(交換樹脂) cdumn chromatography법(法)에 의(依)해서 식용해조(食用海藻)인 '미역'의 단백성질소획분(蛋白性窒素劃分) 및 비단백성질소획분(非蛋白性窒素劃分)의 amino산함량(酸含量)을 정량(定量)하였다. 1. 단백성획분(蛋白性劃分)의 질소함량(窒素含量) (34.1mg/10g)은 비단백성획분질소(非蛋白性劃分窒素)(13i3mg/10g)의 약(約) 10배(倍)이다. 2. 양자(兩者)의 amino산조성(酸組成)은 서로 대조적(對照的)이고 그중(中) 단백성획분(蛋白性劃分)에는 필수(必須) amino산(酸)들이 고루 함유(含有)되어 있으며 그들 함유율(含有率)은 대두단백(大豆蛋白)의 경우(境遇)보다 높았다. 3. 비단백성획분(非蛋白性劃分) (80% ethanol가용(可溶))에는 상당량(相當量)의 Citrulline이 함유(含有)되어 있는것 이는 Ion 교환수지(交換樹脂) Column에서의 용리순위(溶離順位), 일차원(一次元) 및 이차원(二次元) paper chromatogram 상(上)에서의 위치(位置), 그리고 Ninhydrin 및 Ehrlich 시약(試藥)에 대(對)한 양성반응(陽性反應)에 의(依)해서 증명(證明)하였으나 그 정확(正確)한 함량(含量)은 150-cm column에서 Glycine과 중첩(重疊)되어서 용리(溶離)되는 관계(關係)로 알수 없고, 다만 이들 혼합물(混合物)의 질소량(窒素量)이 이획분전질소중(劃分全窒素中) 약(約) 30%에 상당(相當)함을 밝혔다. 4. 비단백성획분(非蛋白性劃分)에서 유리상태(遊離狀態)로 존재(存在)하는 새로운 peptide를 분리(分離)해 냈는데, 이는 염산가수분해후(鹽酸加水分解後) paper chromatography에 의(依)해서 조사(調査)한 바 Alanine, Gluatmic acid 및 Aspartic acid로 조성(組成)된 Tripeptide 임을 알았다. 5. '미역'(건조물(乾燥物))에는 약(約) 1%에 해당(該當)하는 다량(多量)의 Alanine이 함유(含有)되어 있는데 그 중(中) 약(約) 1/3량(量)이 유리상태(遊離狀態)로 존재(存在)함을 알았다.

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콩고오지 제조중(製造中)의 peptide에 관(關)한 연구(硏究) -제3보(第三報) 콩고오지 제조중(製造中)에 생성(生成)되는 저급(低級) peptide의 구조(構造)- (Studies on peptide during soybean-koji preparation -Part III Amino acid sequence of oligopeptides formed during soybean-koji preparation-)

  • 김재욱
    • Applied Biological Chemistry
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    • 제6권
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    • pp.107-117
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    • 1965
  • (1) 콩고오지 제조중(製造中) 생성(生成)되는 저급(低級) peptde의 종류(種類)를 구명(究明)하는 동시(同時)에 겸(兼)하여 Aspergillus soya protease의 작용(作用) specificity를 구명(究明)하기 위(爲)하여 콩고오지 제조중(製造中) 생성(生成)되는 저급(低級) peptide의 N-terminal 및 C-terminal을 동정(同定)하고 dipeptide 및 tripeptide에 대(對)하여서는 amino acid sequence를 결정(決定)하여 다음과 같은 결과(結果)를 얻었다. Gly, Glu. Ala. Ser. Glu. Ser. Ala. Val (Cys, Glu, Ser, Ala, Arg, Try, Leu or Ileu) Asp. Phe (His, Arg, Cys, Asp, Ser, Ala, Leu or Ileu) Glu. Ala (Cys, Gly, Met) Glu. Ala (Asp, Glu,) Gly. Met (Asp, Glu, Ala, Tyr, Leu or Ileu, Lys,) Gly. Leu or Ileu (His, Asp, Glu, Gly, Ser, Lys, Thr, Phe,) Cys. Gly (Asp, Tyr,) Glu. Pro (Asp, Glu, Ser, Gly, Thr, Ala, Val, Leu or Ileu) Try. Ser (Gly, Glu, Arg, Ala, Met, Leu or Ileu,) Asp. Met (Asp, Glu, Ala, Try, Pro, Leu or Ileu,) His Thr (Ser, Gly, Tyr, Pro, Leu or Ileu,) Glu. Gly (Asp, Ala, Ser, Glu,) Leu or Ileu (2) Aspergillus soya protease의 작용(作用) specificity는 비교적(比較的) 광범위(廣範圍)하며 carboxyl기(基)를 가진 amino acid가 acidic amino acid일 때 잘 작용(作用)하는 것으로 보인다.

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담배식물체에서 필수아미노산인 lysyl-glutamyl-tryptophan을 암호화하는 인공유전자의 발현 (Expression of an artificial gene encoding a repeated tripeptide lysyl-g1utamyl-tryptophan in Tobacco Plant)

  • 이수영;나경수;백형석;박희성;조훈식;이용세;최장원
    • 생명과학회지
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    • 제12권1호
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    • pp.96-105
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    • 2002
  • 식물 단백질의 영양가 향상을 위한 일환으로 필수아미노산의 조성이 풍부한 인공단백질을 암호화하는 인공유전자를 담배 식물체에서 발현을 시도하기 위하여, 식물에서 외래유전자의 발현에 널리 사용되는 Cauliflower mosaic virus (CaMV)의 35S promoter를 이중으로 중첩되도록 하고, (Lys-Glu-Trp)이 64번 반복되는 인공유전자 및 nopaline synthase (nos) terminator를 갖고있는 binary vector pART4-4를 구성하였다. 이 재조합 플라스미드는 Agrobacterium tumefaciens를 이용한 형질전환에 의해 Nicotiann tabacum (Var. Xanthi)으로 도입되었다. Kanamycin이 포함된 신초 유도 배지 및 뿌리 유도배지를 이용하여 정상적으로 재생된 담배 식물체로부터 도입된 인공유전자의 발현을 분석하였다. 추출한 genomic DNA를 EcoRI으로 자른 다음 Southern blot 분석에 의하면, 효소 절단 시 예상되는 1.1 kb에서 band를 형성하였으며 각각의 형질전환 식물체에 인공유전자가 1 또는 3 개씩 도입되어 있음을 확인하였다. Northern blot 분석에 의하면 약 1.2 kb 전사체가 비교적 안정하게 발현되었으며, 잎, 줄기, 뿌리로부터 RNA를 분리하여 promoter의 조직 특이성 발현을 분석한 결과, 잎에서 생성되는 RNA가 줄기나 뿌리 조직보다 안정하게 발현되었다. 형질전환 식물체에서 Western blot에 의한 단백질 분석 결과, 잎에서 추출한 단백질로부터 원하는 크기인 33 kDa의 인공단백질이 생성됨을 확인하였으며 발현 수준은 전체 세포 단백질의 0.1%로서 낮은 수준이었다.