• Title/Summary/Keyword: artificial polypeptide

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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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    • v.2 no.1
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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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Flow Characteristics of Elastin-Like Polypeptide(ELP) Coated Packed Bed (온도 감응성 폴리펩티드로 표면 처리된 충전층의 유동 특성)

  • Kim, Duck-Jong;Lee, Jong-Hwan;Park, Sang-Jin;Hyun, Jin-Ho
    • Proceedings of the KSME Conference
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    • 2007.05b
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    • pp.3187-3190
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    • 2007
  • Elastin-Like polypeptide (ELP) composed of elastin-based repeating units is an artificial biomaterial which is biocompatible and non-immunogenic. ELP shows a characteristic inverse phase transition between hydrophobic and hydrophilic phase by external stimuli such as salt, pH and temperature. In this study, ELP coated PS (polystyrene) beads are packed in tubing and the thermo -responsive flow characteristics of the packed bed are investigated. Preliminary test results show that the control of the fluid flow can be achieved by using the temperature driven phase transition effect of the ELP coated beads in a microchannel.

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SYNTHESIS OF BLOCK COPOLYMERS CONTAINING POLYPEPTIDE AND ITS BIOMEDICAL APPLICATION

  • Cho, Chong-Su
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 1997.06a
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    • pp.159-169
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    • 1997
  • ABA-type(or AB) block copolymers composed of poly(${\gamma}$-alkyl L-glutamate) (PALG)[or poly(L-leucine)] as the A component and polyether[or poly (N-isopropy1 acrylamide) as the B component were synthesized by polymerization of (${\gamma}$-alkyl L-glutamate N-carboxyanhydride initiated by primary amined located at both(or one) ends of the polymer chains. Structural studies of the block copolymers were performed in the solution and solid state. Also, artificial skin, drug delivery system of the block copolymers and cell attachment onto the copolymer were carried out for biomedical applications.

Studies on leucocytozoonosis of chickens in Honam districts (호남지방의 닭 Leucocytozoon증에 관한 연구)

  • Rim, Byung-moo;Suh, Min-seuk;Rhee, Jae-ku;Lee, Joo-muk;Kim, Young-jin;Choi, In-yeol;Han, Kyu-sam
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.135-139
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    • 1994
  • In the year 1992/93 leucocytozoonosis could be first diagnbosed in 87 chickens of 4 chicken farms in Honam districts. The diagnosis was confirmed by detection of the blood merozoites or gametocytes and histological finding of the schizonts from various organs with some clinical signs. Cases of leucocytozoonosis only occurred from the end of June to the middle of September. Artificial infection could be observed by means of inoculation of infected blood merozoites. The schizonts were found in the liver and cardiac muscle of the different chickens recovered from the natural infection, respectively, in September and next February. Thus the relapse or long-term infection in cold seasons might be possible. The unique gametocyte antigen polypeptide was 50.1 kD.

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Heterologous Expression of Daptomycin Biosynthetic Gene Cluster Via Streptomyces Artificial Chromosome Vector System

  • Choi, Seunghee;Nah, Hee-Ju;Choi, Sisun;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1931-1937
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    • 2019
  • The heterologous expression of the Streptomyces natural product (NP) biosynthetic gene cluster (BGC) has become an attractive strategy for the activation, titer improvement, and refactoring of valuable and cryptic NP BGCs. Previously, a Streptomyces artificial chromosomal vector system, pSBAC, was applied successfully to the precise cloning of large-sized polyketide BGCs, including immunosuppressant tautomycetin and antibiotic pikromycin, which led to stable and comparable production in several heterologous hosts. To further validate the pSBAC system as a generally applicable heterologous expression system, the daptomycin BGC of S. roseosporus was cloned and expressed heterologously in a model Streptomyces cell factory. A 65-kb daptomycin BGC, which belongs to a non-ribosomal polypeptide synthetase (NRPS) family, was cloned precisely into the pSBAC which resulted in 28.9 mg/l of daptomycin and its derivatives in S. coelicolor M511(a daptomycin non-producing heterologous host). These results suggest that a pSBAC-driven heterologous expression strategy is an ideal approach for producing low and inconsistent Streptomyces NRPS-family NPs, such as daptomycin, which are produced low and inconsistent in native host.

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

  • Lee, Soo-Young;Ra, Kyung-Soo;Baik, Hyung-Suk;Park, Hee-Sung;Cho, Hoon-Sik;Lee, Young-Se;Choi, Jang-Won
    • Journal of Life Science
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    • v.12 no.1
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    • pp.96-105
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    • 2002
  • To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.

Expression and Purification of an ACE-Inhibitory Peptide Multimer from Synthetic DNA in Escherichia coli

  • OH, KWANG-SEOK;YONG-SUNG PARK;HA-CHIN SUNG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.59-64
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    • 2002
  • An angiotensin I-converting enzyme (EC 3.4.15.1) (ACE), which can convert inactive angiotensin I into angiotensin II, a vasoconstrictor, is one of the key enzymes in controlling hypertension. It is suggested that the inhibition of ACE prevents hypertension, and many inhibitory peptides have already been reported. In the current study, oligonucleotides encoding ACE inhibitory peptides (IY, VKY) were chemically synthesized and designed to be multimerised due to isoschizomer sites (BamHI, BglII). The cloned gene named AP3 was multimerised up to 6 times in pBluescript and expressed in BL2l containing pGEX-KG. The fusion protein (GST-AP3) was easily purified with a high recovery by an affinity resin, yielding 38 mg of synthetic AP3 from a 1-1 culture. The digestion of AP3 by chymotrypsin exhibited an $IC_50$ value of $18.53{\mu}M$. In conclusion, the present experiment indicated that AP3 could be used as a dietary antihypertensive drug, since the potent ACE inhibitory activity of AP3 could be activated by chymotrypsin in human intestine.

Effect of Serum IGF-I on Progesterone Concentration during Early Pregnancy in Korean Native Cattle (Hanwoo)

  • Ryu, Yang-Hwan;Yang, Jong-Yong;Seo, Dong-Sam;Ko, Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.176-179
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    • 2003
  • Insulin-like growth factor-I (IGF-I) is a polypeptide that has the function of regulating the expression of steroid hormones through endocrine, paracrine, and autocrine actions in reproductive organs. Moreover, IGF-I is involved in ovulation, implantation, maintenance of pregnancy, and development of fetuses in animals. Therefore, this study was conducted to investigate the effects of serum IGF-I concentration on progesterone ($P_4$) concentration and pregnancy rates in Korean native cattle (Hanwoo). Blood was collected at estrus (Day 0) and Day 11. Artificial insemination was performed at Day 0. Serum IGF-I and $P_4$ concentrations were measured by radioimmunoassay (RIA). Overall, $P_4$ concentration was higher at Day 11 than Day 0, whereas the pattern of IGF-I concentration was reversed. When animals were divided into two groups depending on the pregnancy status, $P_4$ concentrations of the pregnant group was significantly higher than that of the non-pregnant group at Day 0 (p<0.05) and Day 11 (p<0.05). But, lower IGF-I concentrations were detected in the pregnant group at Day 0 (p<0.05) and Day 11 (p<0.05) compared to the non-pregnant group. In conclusion, these results indicated that serum IGF-I is inversely associated with $P_4$ concentration during early pregnancy in Hanwoo.