• Title/Summary/Keyword: Functional Amino Acids

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Cloning, Expression, and Functional Characterization of the Dunaliella salina 5-enolpyruvylshikimate-3-phosphate Synthase Gene in Escherichia coli

  • Yi, Yi;Qiao, Dairong;Bai, Linhan;Xu, Hui;Li, Ya;Wang, Xiaolin;Cao, Yi
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.153-157
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    • 2007
  • 5-enolpyruvylshikimate-3-phosphate synthase (EPSP synthase, EC 2.5.1.19) is the sixth enzyme in the shikimate pathway which is essential for the synthesis of aromatic amino acids and many secondary metabolites. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide glyphosate. In this study, the Dunaliella salina EPSP synthase gene was cloned by RT-PCR approach. It contains an open reading frame encoding a protein of 514 amino acids with a calculated molecular weight of 54.6 KDa. The derived amino acid sequence showed high homology with other EPSP synthases. The Dunaliella salina EPSP synthase gene was expressed in Escherichia coli and the recombinant EPSP synthase were identified by functional complementation assay.

Molecular Analysis of Korean Isolate of Barley mild mosaic virus (Iks Isolate)

  • Choi, Min-Kyung;Kamala-Kannan, Seralathan;Oh, Byung-Taek;Park, Jong-Chul;Lee, Gun-Woong;Lee, Kui-Jae;Park, Yool-Jin
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.157-164
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    • 2009
  • The complete nucleotide sequences of both RNA of an isolated Barley mild mosaic virus (BaMMV) from Iksan, Korea, have been determined. RNA1 was 7273 nucleotides long and encodes for a polyprotein of 2261 amino acids, which contains the eight putative functional proteins. RNA2 was 3520 nucleotides long and encodes for a polyprotein of 894 amino acids, which contains two functional proteins. Results of multiple sequence alignment showed 92.9% similarity with Na1 isolate, followed by Sil, UK(F), Asl1, Remis M and UK(M) isolates, respectively. Comparison of the BaMMV-Iks polyproteins with the corresponding proteins of BaMMV-Na1 isolates showed 95% amino acid sequence identity. The phylogenetic analysis revealed that Iks isolate was closely related to Na1 strain and have a common origin.

Functional Ingredient Compositions of Soybean Curds(Tofu) Made with Black Soybeans(Huktae) and White Soybeans(Baktae) (백태와 흑태를 원료로 하여 제조된 두부의 기능성 성분 분석)

  • Kim, Kang-Sung
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.158-163
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    • 2007
  • The compositions of tofus made with Baktae and Huktae were compared to those made with other respective types of soybeans. The lipid and protein contents of Baktae were 20.8% and 39.7%, while those of Huktae were 15.8% and 41.1%, respectively. In both soybean types, linoleic acid(18:2), oleic acid(18:1), and palmitic acid(16:0) accounted for more than 80% of the total fatty acids. The tofu made from Baktae was whiter than the tofu made from Huktae, as exhibited by a higher Hunter's L-value. The lipid content of the Baktae tofu(4.41%) was higher than that of the Huktae tofu(3.26%). The amino acid compositions of the Huktae and Baktae tofus were similar, with glutamic acid and aspartic acid being the most abundant amino acids in both tofus. However, the content of the limiting amino acid, methionine, increased 2.5-fold in the tofus as compared to the soybeans, on a per gram protein basis. Isoflavones were lost during tofu making as contents for the tofu were 20${\sim}$25% of those for the soybeans.

Simultaneous GC/MS Analyses of Organic acids and Amino acids in Urine using TMS-TFA derivative (TMS-TFA 유도체화를 이용한 소변여지 중 유기산과 아미노산의 GC/MS 동시분석)

  • Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.19 no.1
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    • pp.107-114
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    • 2006
  • Early diagnosis and medical intervention are critical for the treatment of patients with metabolic disorders. A rapid analytical method was developed for simultaneous quantification of organic acids and amino acids in urine without labor-intensive pre-extraction procedure showing high sensitivity and specificity. A new method consisted of simple two-step trimethylsilyl (TMS)-trifluoroacetyl (TFA) derivatization using GC/MS-selective ion monitoring (SIM). Filter paper urine specimens were dried under nitrogen after being fortified with internal standard (tropate) in a mixture of distilled water and methanol. Methyl orange was added to the residue as indicator reagent. Silyl derivative of carboxylic functional group was followed by trifluoroacetyl derivative for amino functional group. N-methyl-N-(trimethylsilyl-trifluoroacetamide) and N-methyl-bistrifluoroacetamide were consecutively added and heated for 15-20 min at $65^{\circ}C-70^{\circ}C$, for TMS-TFA derivative, respectively. This reactant was analyzed by GC/MS-SIM. Linear dynamic range showed 0.001-50 mg with the detection limit of (S/N=3) 10-200 ng, and the quantification limit of 80-900 ng in urine. Correlation coefficient of regression line was 0.994-0.998. When the method was applied to the patients 'urine, it clearly differentiated the normal from the patient with metabolic disorder. The study showed that the developed method could be the method of choices in rapid and sensitive screening for organic aciduria and amino acidopathy.

Cloning of the Transketolase Gene from Erythritol-Producing Yeast Candida magnoliae

  • Yoo, Boung-Hyuk;Park, Eun-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1389-1396
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    • 2014
  • The entire nucleotide sequence of the TKL1 gene encoding transketolase (TKL) in an erythritol-producing yeast of Candida magnoliae was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed an open reading frame of C. magnoliae TKL1 (CmTKL1) that spans 2,088 bp and encodes 696 amino acids, sharing 61.7% amino acid identity to Kluyveromyces lactis TKL. Functional analysis showed that CmTKL1 complemented a Saccharomyces cerevisiae tkl1 tkl2 double mutant for growth in the absence of aromatic amino acids and restored transketolase activity in this mutant. An enzyme activity assay and RT-PCR revealed that the expression of CmTKL1 is induced by fructose, $H_2O_2$, and KCl. The GenBank accession number for C. magnoliae TKL1 is KF751756.

Varietal Comparison of Composition Characteristics in Several Mulberry Leaves Produced in Korea (한국산 품종별 뽕나무 잎(桑葉)의 화학적 성분 특성)

  • 이성갑;천동현;선종연
    • Journal of the Korean Professional Engineers Association
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    • v.34 no.3
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    • pp.68-73
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    • 2001
  • This study was conducted to investigate the possible use of Mulberry leaf as a functional supplement for Mulberry leaves. Chemical characteristics of mulberry leaf were investigated in eleven mulberry varieties, including Suwon, Chongil, Gumseol, Sugae, Yongchun, Hongol, Daeryk, Kaerang, Donae, Chungol and Kuksang20 varieties. The chemical compositions of minerals, amino acids, rutin as flavonol glucoside were analyzed by AOAC methods, HPLC method, amino acid analyzer, respectively. The results are as follows: In case of KukSang Mulberry leaf with spring harvest, the content of general components was crude protein 16.28%, carbohydrate 69.12%, crude fiber 9.15%, crude ash 8.44% and crude fat 2.57% respectively. It is noteworthy that ferrious content of KukSang mulberry leaves contains twice as great as that in other variety of mulberry leaves. The content of alanine was the highest among amino acids in all variety mulberry leaf. Spring leaf contain more utin content than fall leaf, among the variety, spring HongAll leaf rutin contain 260mg/100g as higher than that of fall ChungAll leaf 70mg/100g).

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A Study on Development of Protein Materials using Dead Flatfish from Fish Farms(1) -Antioxidant Functional Properties- (양식장 넙치 폐사어를 이용한 단백질 소재의 개발에 관한 연구(1) -항산화 기능특성-)

  • Kang, Keon-Hee;Lee, Min-Gyu;Kam, Sang-Kyu;Jeong, Kap-Seop
    • Journal of Environmental Science International
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    • v.22 no.11
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    • pp.1465-1471
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    • 2013
  • To develop the protein materials by the reutilization of dead flatfish from fish farms in Jeju island, the physicochemical characteristics and the functional activities of collagen peptide extracts were investigated. Flatfish skin collagen peptide (FSCP) and flatfish protein hydrolysate (FPH) were manufactured from dead flatfish. The differences of pH, moisture and fat contents between FSCP and FPH were not significant, fat contents were analyzed less than 0.3%, and trans-fat, saturated fat and cholesterol were not detected in both samples. Protein contents of FSCP and FPH showed about 92% and 95%, respectively. In the analysis of amino acids, glycine and hydroxy proline content in FSCP was 24.22% and 6.15%, respectively, showed a typical characteristics of the collagen protein, but essential amino acids contents such as threonine, valine, methionine, isoleusine, leusine and phenylalanine were relatively higher than those of FPH. Average molecular weight of FSCP was measured as 1,102 which was almost equal value with that of tuna collagen peptide. The antioxidant activities and functional properties showed high but did not show significant difference between two samples.

Identification of an Embryonic Growth Factor IGF-II from the Central Nervous System of the Teleost, Flounder, and Its Expressions in Adult Tissues

  • Kim, Dong-Soo;Kim, Young-Tae
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.113-118
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    • 1999
  • The insulin-like growth factor (IGF) is found in all vertebrates and its type-II molecule is regarded as a fundamental embryonic growth factor during development. We have firstly identified, in this study, a cDNA clone corresponding to IGF-II (flIGF-II) from the adult brain of the teleost, Paralichthys olivaceus. We also examined the tissue expression of flIGF-II in several adult tissues by RT-PCR. The flIGF-II cDNA contained a complete ORF consisting of 215 amino acids and one stop codon. Its molecular characteristics appear to be similar to the previously identified IGF-II molecules, in which a common primary structure exhibiting B, C, A, D, and E domains is evidently observed. This cDNA clone seems to be cleaved at $Ala_{52}$ for the $NH_2$-end signal peptide and appears to produce a 98 amino acid-long E-peptide from the $Arg^{118}$. The functional B-D domain regions, therefore, include 65 amino acids and is able to encode a 7.4-kDa protein. The most prominent structural difference between IGF-I and IGF-II was that the D domain of IGF-II exhibits a two-codon-deleted pattern compared to the 8 amino acid-containing IGF-I. The insulin family signature in the A domain and six cysteins forming three disulfide bridges between the B and A domains were evolutionary-conserved from teleosts to mammalian IGF-II. Interestingly, the E-peptide region appears to provide a distinct hallmark between teleosts in amino acid composition. The flIGF-II shows 85.1% of sequence identity to salmon and trout, 90.6% to tilapia, and 98.4% to perch in amino acid level. In tissue expressions of IGF-II, it is very likely that flIGF-II has a significant expression in the adult brain. However, liver seems to be the main source for IGF-II production, and relatively low signals were observed in the adult muscle and kidney. Taken together, it would be concluded that the functional region for IGF-II mRNA is highly similar in phylogeny and is evolutionary, conserved as a mediator for the growth of vertebrates.

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Physicochemical Properties of the Turmeric (Curcuma longa L.) in Jindo Korea (진도산 울금(Curcuma longa L.)의 이화학적 특성)

  • Oh, Da-Young;Kim, Han-Soo
    • Journal of Environmental Science International
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    • v.28 no.4
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    • pp.403-412
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    • 2019
  • Studies on the physicochemical properties of physiological activity substance in turmeric (Curcuma longa L.) were analyzed for the use as an functional food materialization. The proximate compositions in the vacuum freeze dried turmeric were carbohydrate 72.90%, moisture 5.74%, crude protein 10.02%, crude fat 4.67%, and crude ash 6.69%, respectively. The mineral contents of turmeric were calcium (Ca) $2,294.77mg\;kg^{-1}$, potassium (K) $28,780.54mg\;kg^{-1}$, magnesium (Mg) $2,826.90mg\;kg^{-1}$, sodium (Na) $1,826.58mg\;kg^{-1}$, iron (Fe) $190.94mg\;kg^{-1}$, and manganese (Mn) $620.16mg\;kg^{-1}$. The vitamin contents of turmeric were pantothenic acid 1.040 mg/100 g, riboflavin 0.166 mg/100 g, thiamin 0.148 mg/100 g, pyridoxine 0.010 mg/100 g, and calciferol 0.008 mg/100 g, respectively. Total amino acid contents in protein of turmeric were 7.66 g%, and major amino acids were aspartic acid 1.45 g%, glutamic acid 1.07 g%, leucine 0.71 g%, phenylalanine 0.47 g%, and arginine 0.46 g%, respectively. The amount of free amino acids of turmeric were 225.81 mg%, and major free amino acids were asparagine, aspartic acid, glutamic acid, serine, and alanine. Especially, in the case of asparagine, it was highest. The compositions of fatty acid were saturated fatty acid 45.09%, monoenes 8.62%, and polyenes 46.30%.

Physicochemical attributes, oxidative stability, and microbial profile of boneless sirloin and bone-in T-bone steaks from Hanwoo steer with reference to dry-aging

  • Ali, Mahabbat;Nam, Ki-Chang
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1169-1181
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    • 2021
  • We investigated the comparative physicochemical attributes, oxidative stability, and microbial characteristics of 28 days dry-aged meat in between boneless sirloin (gluteus medius) and bone-in T-bone steaks (infraspinatus) muscles from Korean Native Hanwoo Steer (KNHS). Results reveal that regardless of the muscles, dry-aging increased protein content and water-holding capacity (WHC) (p < 0.05). Meat from infraspinatus-aged muscle led to darker meat with higher pH values than un-aged meat (p < 0.05). However, fat content, CIE a*, and CIE b* remained unchanged in both muscles at aging. At aged meat, thiobarbituric acid reactive substances (TBARS) values from bone-in infraspinatus muscle was 2.5-fold higher than boneless gluteus medius muscle (p < 0.05). Dry-aging led to an increase in the contents of total unsaturated fatty acids (UFAs), monounsaturated fatty acids (MUFAs), and UFA/saturated fatty acids (SFA) in both muscles (p < 0.05). Furthermore, gluteus medius aged muscle concentrated with olic acid (C18:1) compared to infraspinatus aged muscle. Irrespective of the muscles, dry-aging enhanced the total free amino acids (FAAs) as well as tasty, and bitter amino acid contents whereas decreased the tasty/bitter amino acids (p < 0.05). Aromatic amino acid, tryptophan that converted to serotonin was 2-fold higher in boneless gluteus medius muscle than bone-in infraspinatus muscle at pre and post aging processes (p < 0.05). Aged Infraspinatus muscle increased total bacteria (p < 0.05) while no salmonella spp. was detected in both muscles. Taken together, our study confirms that 28 days dry-aging profiling the quality characteristics of boneless sirloin (gluteus medius) and bone-in T-bone steaks (infraspinatus) distinctly while gluteus medius aged steak performs better owing to oxidative stability and functional compounds than infraspinatus aged steak.