• Title/Summary/Keyword: amino acid substitution

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Microwave Assisted Synthesis of 1,3,4-Oxadiazole/Thiohydantoin Hybrid Derivatives via Dehydrative Cycliztion of Semicarbazide

  • Yang, Seung-Ju;Lee, Jae-Min;Lee, Gee-Hyung;Kim, NaYeon;Kim, Yong-Sang;Gong, Young-Dae
    • Bulletin of the Korean Chemical Society
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    • v.35 no.12
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    • pp.3609-3617
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    • 2014
  • A series of compounds containing both 1,3,4-oxadiazole and thiohydantoin were synthesized as a promising scaffold for application in medicinal chemistry. The key step of the synthesis is a microwave-assisted cyclization of semicarbazides possessing a thiohydantoin moiety at one of the acyl termini using $POCl_3$ as a dehydrating reagent. A wide range of semicarbazides were prepared through the substitution of hydrazides with an N-acylated thiohydantoin derived from the cyclization of the corresponding isothiocyanate with an amino acid and subsequent N-acylation of the resultant thiohydrantion. Consequently, the 58 number of 1,3,4-oxadiazole derivatives having a thiohydantoin substituent were prepared in good overall yields.

Improvement of Bacterial Endo-1,4-,\beta-D-glucanase(CMCase) Secretion in Yeast by Mutagenesis of Glucoamylase Signal Sequence. (Glucoamylase 분비신호서열의 돌연변이에 의한 효모에서 세균의 Endo-1,4-\beta-D-glucanase의 분비능 증진)

  • 이준원;강대욱;김보연;오원근;민태익;이상원;변유량;안종석
    • Microbiology and Biotechnology Letters
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    • v.28 no.4
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    • pp.195-201
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    • 2000
  • Glucoamylase of Saccharomyces diastaticus is produced as a large precursor composed of signal peptide (21 amino acid residues), Thr and Ser-rich region and functional glucoamylase. To evaluate the utility of the glucoamylase signal peptide (GSP) for the secretion of foreign proteins, four types of GSP mutants (ml : Pro-18 longrightarrowLeu-18, m2 : Tyr-13 longrightarrowLeu, m3 : Ser-9longrightarrowLeu-9, m4 : Asn-5 longrightarrowPro-5) were constructed and secretion efficiency of each mutant was compared with that of native GSP by the expression and secretion of Bacillus subtilis CMCase under the control of GAP in N-terminal domain and hydrophobic domain. n mutant 4, a polar amino acid was replaced by a helix - breaking Pro residue. CMCase activity assay and Western blot analysis revealed that CMCase secretion by GSP mutants replaced by Leu were increased compared with native GSP. In the case of m2 and m3, the substitution of Leu for Tyr-13 and Ser-9 in the hydrophobic region resulted in a twofold increase in the extracellular CMCase activity.

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Identification of Amino Acid Residues Involved in Feedback Inhibition of the Anthranilate Synthase in Escherichia coli

  • Kwak, Joon-Hyeok;Hong, Kwang-Won;Lee, Sung-Haeng;Hong, Jin-Han;Lee, Se-Yong
    • BMB Reports
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    • v.32 no.1
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    • pp.20-24
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    • 1999
  • The first step of the branch pathway in tryptophan biosynthesis is catalyzed by anthranilate synthase, which is subjected to feedback inhibition by the end product of the pathway. The $trpE^{FBR}$ gene from a mutant Escherichia coli strain coding for anthranilate synthase that was insensitive to feedback inhibition by tryptophan has been cloned. To identify the amino acid changes involved in the feedback regulation of anthranilate synthase, the nucleotide sequence of the mutant $trpE^{FBR}$ gene was determined. Sequence analysis of the $trpE^{FBR}$ gene revealed that four bases were changed in the structural gene while alteration was not found in the 5' control region. Among these base changes, only two base substitutions caused the alterations in amino acid sequences. From the results of restriction fragment exchange mapping, the 61st nucleotide, C to A substitution, that changed $Pro^{21}{\rightarrow}Ser$ was identified as the cause of the desensitization to feedback inhibition by tryptophan. Additional feedback-resistant enzymes of the E. coli anthranilate synthases were constructed by site-directed mutagenesis to examine the effect of the $Ser^{40}\;{\rightarrow}\;Arg^{40}$ change found in the $trpE^{FBR}$ gene of Brevibacterium lactofermentum. From the feedback inhibition analysis, the $Pro^{21}{\rightarrow}Ser$ and $Ser^{40}{\rightarrow}Arg$ mutants maintained about 50% and 90% of their maximal activities, respectively, even at the extreme concentration of 10 mM tryptophan. From these results, we suggest that the $Pro^{21}$ and $Ser^{40}$ residues are involved in the tryptophan binding in the E. coli enzyme.

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Effects of Oatrim on the Quality of Meat Products (일부재료(Oatrim)대체가 육류제품의 품질에 미치는 영향)

  • Jeon, Yoo-Shin;Sohn, Kyung-Hee
    • Korean journal of food and cookery science
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    • v.10 no.3
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    • pp.254-259
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    • 1994
  • Meat provides high quality proteins, lipids, minerals and vitamins. The meat protein is especially high in essential amino acids that are crucial for human health, growth & development and for the formation of enzymes, hormones and antibodies. Relatively cheap and nutritionally sound vegetable proteins that are similar to animal proteins are being developed to replace the animal proteins in texture, nutrition and food characteristics. In this study a nutritionally sound meat lipid replacing food Oatrim that has been produced by converting oat starch into maltodextrin by ${\alpha}$-amylase, have been partially substituted for beef and general component analysis, texture measurement and sensory tests have been conducted. The results are 1. Water content of the non-treated (0% treated) was 67.1% and the treated (10% treated) was 77%. The treated showed better water holding capacity. 2. Protein content of the non-treated was 21.2 g/100 g; the 4% treated, 18.4 g/100 g; the 6% treated, 18.2 g/100 g; the 8% treated, 17.2 g/100g; and the 10% treated, 16.0 g/100 g. The protein content tended significant. 3. Amino acid analysis results showed that glutamic acid content was the highest in Oatrim and as its amino acid make up is exellent, it is valuable as a fine low fat protein food. 4. Sensory tests show that the increased Oatrim content increased the appearance quality but food characteristics were high only in the 4% and 6% treated groups, indicating that the replacement ratio should not exceed 10%. 5. Texture measurement analysis results show that the higher the replacement content, lower the springness, cohesiveness, hardness, chewiness and gumminess, resulting in relatively soft overall texture. However, in order to better the food characteristics, more studies must be continuously done, and so by being able to increase vegetable substitution over meat, it may be able to contribute to the prevention of adult disease.

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The Complete Amino Acid Sequence of Newborn Dog Prochymosin (강아지 프로카이모신의 전 아미노산 서열)

  • Yoon, Joo-Ok;Kim, Hyun-Ku
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.3
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    • pp.289-300
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    • 1997
  • Newborn dog chymosin was extracted from the stomachs of dogs of 2 weeks of age, and was purified by ion exchange chromatography. Half of the sequence was determined by amino acid sequencing and the complete sequence was deduced from a cloned chymosin cDNA Results showed that the zymogen showed 79% sequence identity with calf prochymosin and 54% identity with porcine pepsinogen A The size of the propart and location of the residue which becomes the amino-terminus in the active enzyme was the same in the prochymosins. The maximum general proteolytic activity at pH 3.2 of newborn dog chymosin was 3-4% of that of porcine pepsin A at pH 2, whereas the milk clotting activity relative to the general proteolytic activity of newborn dog chymosin was much higher than that of calf chymosin. Agar gel electrophoresis at pH 5.2 of stomach extracts of individual dogs showed the existence of two predominant genetic variants of zymogen and enzyme. The two variants could not be distinguished by amino acid composition or amino-terminal sequencing, and no differences in the enzymatic properties of the genetic variants were observed. It was concluded that of the residues that participate in the substrate binding, calf and newborn dog chymosin differ in the following positions (porcine pepsin numbering, subsites in parentheses) : Ser 12 Thr(S$_4$), Leu 30 Val(S$_1$/S$_3$), His 74 Gln(S'$_2$), Val 111 Ile(S$_1$/S$_3$), Lys 220 Met(S$_4$). With regard to the low general proteolytic activity of newborn dog chymosin, the substitution Asp303 Val relative to calf chymosin may contribute to an explanation of this.

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Preparation of High Range Water Reducer Containing Carboxylic Acid and Their Cement Absorptivity(I) (카르본산계 고성능감수제의 제조 및 그들의 시멘트 흡착성(I))

  • 김화중;강인규;김성훈;김우성;권영도
    • Magazine of the Korea Concrete Institute
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    • v.7 no.2
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    • pp.175-182
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    • 1995
  • Styrene-Maleic anhydride copolymer (SMA) was prepared by the radical copolmerization of styrene and maleic anhydride using ${\alpha}-{\alpha}'$ azobis(isobutyronitrile) as an initiatrr. SMA was further reacted with m-amino phenol to obtain aminophenol-substituted SMR (mSMA). Sulfonated SMA and mSMA were also prepared by the reaction of copolymers with sulfuric acid The copolyniers were characterized by infrared spectroscopy. It was found from the results of elemental analysis that the substitution degree of aminophenol in the mSMR is 44% and the degree is lowered to 35% after sulfonation. The percentage of copolymers adsorbed on the surface of cement particles was increased with a decrease of added copolymers. While, the arnourit of sulfonated SMA absorbed on the surface of cement particles was larger than that of the sulfonated mSMA The copolymers synthesized in this study are probably expected as a high range water reducer for coiicxte.

Cloning of the Adenosine Deaminase Gene from Pseudomonas iodinum IFO 3558

  • Jo, Young-Bae;Baik, Hyung-Suk;Bae, Kyung-Mi;Jun, Hong-Ki
    • Journal of Life Science
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    • v.9 no.2
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    • pp.9-14
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    • 1999
  • Pseudomonas iodinum IFO 3558 adenosine deaminase(ADA) gene was cloned by the polymerase chain reaction and deduced the amino acid sequence of the enzyme. DNA sequence homology of Pseudomonas iodinum IFO 3558 ADA gene was compared to those of E. coli, human and mouse ADA genes. Unambiguous sequence from both strands of pM21 was obtained for the region believed to encode ADA. The sequence included a 804-nucleotide open reading frame, bounded on one end by sense primer and on the other end by two antisense primer. This open reading frame encodes a protein of 268 amino acids having a molecular weight of 29,448. The deduced amino acid sequence shows considerable similarity to those of E. coli, mouse and human ADA. Pseudomonas iodinum IFO 3558 nucleotide sequence shows 98.5% homology with that of the E. coli ADA sequence and 51.7% homology with that of the mouse ADA sequence and 52.5% homology with that of the human ADA sequence. The ADA protein sequence of Pseudomonas iodinum IFO 3558 shows 96.9% homology with that of the E. coli and 40.7% homology with that of the mouse and 41.8% homology with that of the human. The distance between two of the conserved elements, TVHAGE and SL(1)NTDDP has veen exactly conserved at 76 amino acids for all four ADAs. Two of the four conserved sequence elements shared among the four ADAs are also present in the yeast, rat, human (M), and Human(L) AMP deaminase. The SLSTDDP sequence differs only in the conservative substitution of a serine for an asparagine. A conserved cysteine with conserved spacing between these two regions is also found. Thus, sequence analysis of four ADAs and four AMP deaminases revealed the presence of a highly conserved sequence motif, SLN(S)TDDP, a conserved dipeptide, HA, and a conserved cysteine residue.

Thermotropic Liquid Crystalline Behavior of Aliphatic Acid Esters of N,O-Hydroxypropyl Chitosans (N,O-히드록시프로필 키토산 지방산 에스터들의 열방성 액정 거동)

  • Kim, Hyo Gap;Jung, Seung Yong;Ma, Yung Dae
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.276-287
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    • 2013
  • Two kinds of N,O-hydroxypropyl chitosans (HPCTOs) with degree of substitution (DS) and molar substitution (MS) ranging from 2.15 to 2.39 and 2.9 to 4.1, respectively, and five kinds of aliphatic acid esters of HPCTOs (HPCTOAms, m=0,2,4,7,9, the number of methylene units in aliphatic substituent) based on the HPCTOs were synthesized, and the thermotropic liquid crystalline properties of the derivatives were investigated. All the derivatives formed enantiotropic cholesteric phases whose optical pitches (${\lambda}_m$'s) increased with increasing temperature. However, the glass and clearing temperatures, the magnitude of ${\lambda}_m$ of the mesophase at the same temperature, and the temperature dependence of ${\lambda}_m$ of the investigated derivatives highly depended on MS and m. The thermotropic mesophase properties of HPCTOAms were significantly different from those reported for the aliphatic acid esters of hydroxypropyl celluloses. The results indicate that the secondary amino group in the C-2 position plays an important role in the thermal stabilization and temperature dependence of ${\lambda}_m$ of the cholesteric mesophase.

Synthesis of Anticoagulant 3-(N-Aryalmino)-1,4-Naphthoquinones(III) (항응고성의 3-(N-Arylamino)-1,4-Naphthoquinone 유도체 합성(III))

  • Ryu, Chung-Kyu
    • YAKHAK HOEJI
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    • v.34 no.6
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    • pp.422-428
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    • 1990
  • 2,3-Dibromo-1,4-naphthoquinone was reacted with p-aminobenzoic acid, 2-aminopyridine, 2-amino-4-metylpyridine, m-nitroaniline, sulfathiazol, p-chloroaniline, phenetidine and 2-bromo-3-(N-arylamino)-1,4-naphthoquinones($1{\sim}8$). 2,3-Epoxy-2,3-dihydro-1,4-naphthoquinone was also reacted with p-amonobenzoic acid, p-toluidine, p-chloroaniline, m-chloroaniline, m-nitroaniline, p-phenetidine, N,N-dimethyl-1,4-pheylenediamine as a ring opening and dehydogenation to form 2-hydroxy-3-(N-arylamino)-naphthoquinones ($9{\sim}16$) in good yield. These new compounds($1{\sim}16$) are expected to have a biological activities such as anticoagulant and cytotoxic.

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Preparation and Characteristics of Fish-frame-added Snacks (Fish-frame을 이용한 snack의 제조 및 특성)

  • Kang Kyung-Tae;Heu Min-Soo;Kim Jin-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.3
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    • pp.261-268
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    • 2006
  • Fish-frames are processing byproducts, which are left after obtaining fillets or muscle during fish processing. The fish-frame generally consists of muscle, collagen, calcium, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). We used fish-frame powder (FFP) of chum salmon and skipjack tuna to prepare and characterize snacks for human consumption with different proportions of FFP. The crude protein and lipid contents of fish-frames were 16.3 and 9.4% for chum salmon and 18.6 and 8.3% for skipjack tuna, respectively. The volatile basic nitrogen (30.6 mg/100 g) and browning index (0.393) of FFP from chum salmon were lower than those of FFP from skipjack tuna. Thus, the FFP of chum salmon was better for making snacks than that of skipjack tuna. Five snacks were prepared with 0, 10, 20, 30, and 40% (w/w) substitution ratios of FFP from chum salmon. The moisture content of the snacks decreased (33.6 to 11.5%) with increasing FFP substitution ratio, whereas crude ash (2.9 to 7.5%), protein (11.4 to 18.4%) and lipid (13.7 to 35.1%) increased. Sensory scores for the texture and taste of the snack with 30% FFP were significantly higher (p<0.05) than those for other snacks; the color and flavor scores of all snacks did not differ significantly. The major fatty acids in the snacks were 16:0 and 18:0 as saturates, 18:1n-9 as monoenes, and 18:2n-6 and 18:3n-3 as polyenes. Snacks with FFP contained small amounts of EPA (0.5 to 0.8%) and DHA (1.3 to 1.8%) in the total lipid composition. The total amino acid content (16.08 g/100 g) of the snack with 30% FFP was higher than that of the snack without FFP (11.18 g/100 g), and the major amino acids were aspartic acid, glutamic acid, glycine, leucine, and lysine. The calcium and phosphorus contents of the snack with 30% FFP were 1,272 mg/100 g and 854 mg/100 g, respectively, and their ratio was the optimal range (2:1 to 1:2) for body absorption efficiency.