• Title/Summary/Keyword: threonine

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Biotechnology for the Production of Threonine Production (Threonine의 생물공학적 생산)

  • Kim, Kyoung-Ja
    • YAKHAK HOEJI
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    • v.34 no.6
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    • pp.447-456
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    • 1990
  • Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

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Effects of Dietary Protein and Threonine Supply on In vitro Liver Threonine Dehydrogenase Activity and Threonine Efficiency in Rat and Chicken

  • Lee, C.W.;Oh, Y.J.;Son, Y.S.;An, W.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.10
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    • pp.1417-1424
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    • 2011
  • This study was conducted to assess the relation between threonine (Thr) oxidation rate and threonine efficiency on rat and chicken fed with graded levels of protein and threonine. The increase in threonine content from 0.28 to 0.72% in a diet containing 12.0% crude protein (CP) caused a gradual increase in threonine dehydrogenase (TDG) activity in rat liver. Similar, but more pronounced results were observed after 18.0% CP in the diet. Both protein levels in combination with the highest level of threonine supplementation increased liver TDG activity significantly, indicating enhanced threonine catabolism. Parameters of efficiency of threonine utilization calculated from parallel nitrogen balance studies decreased significantly and indicated threonine oversupply after a maximum of threonine supplementation. At the lower levels of threonine addition the efficiency of threonine utilization was not significantly changed. In the chicken liver up to 0.60% true digestible threonine (dThr) in the 18.5% CP diet produced no effect on the TDG activity. However, TDG activity in the liver was elevated by the diet containing 22.5% CP (0.60% dThr) and the efficiency of threonine utilization decreased, indicating the end of threonine limiting range. In conclusion, the in vitro TDG activity in the liver of rat and growing chicken has an indicator function for the dietary supply of threonine.

High Production of L-Threonine using Controlled Feeding of L-Methionine and Phosphate by Escherichia coli Mutant (L-Methionine과 Phosphate의 제한 공급에 의한 Escherichia coli MT201로부터의 고농도 L-Threonine 생산)

  • 이만효;이홍원;김병진;김천석;정준기;황용일
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.149-153
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    • 2004
  • L-Threonine fermentation process was constructed on batch and fed-batch culture by using Escherichia coli MT201. The production type of L-threonine was observed as growth-associated production in batch culture. In fed-batch culture studying optimal concentration of yeast extract in feeding media, when 600 g/l of glucose and 60 g/l of yeast extract were added in feeding media, 87 g/$\ell$ of L-threonine was produced. To improve cell growth and L-threonine production, the culture of high cell density was performed in fed-batch culture with oxygen enriched air and feeding media containing L-methionine and phosphate. Under the conditions, we could achieve the highest L-threonine production of98 g/$\ell$ at 60 h. The highest productivity of L-threonine was about 3.85 g/$\ell$/h.

Effect of Protein and Threonine Level in the Diet on Liver Threonine Dehydrogenase Activity in Growing Rat (성장하는 랫트에 있어서 단백질과 Threonine의 급여 수준이 간 Threonine Dehydrogenase 활성에 미치는 영향)

  • 이철원
    • The Korean Journal of Food And Nutrition
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    • v.15 no.3
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    • pp.235-240
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    • 2002
  • The aim of this investigation was to determine the influence of different protein and threonine (Thr) levels on the liver threonine dehydroaenase (TDG) activity b\ulcorner rats. In rats fed on CP (crude Protein) - diets, TDG activity was increased during an CP rise to 12.0% CP, decreased slightly down to 18.0% CP and showed .a trend to increase from 18.0 to 24.0% CP. In rats the feeding with graded Protein supply gavee no indication for additional stimulation of threonine-oxidation by TDG over a wide range of CP-content in the diets. The increase in threonine content from 0.28 to 0.72% in the presence of 12.0% CP caused a gradual increase in TDG activity in rat liver. This similarly applied to the feed admixed with 18.0% CP, but at a higher level.

Development of L-Threonine Producing Recombinant Escherichia coli using Metabolic Control Analysis (대사 조절 분석 기법을 이용한 L-Threonine 생산 재조합 대장균 개발)

  • Choi, Jong-Il;Park, Young-Hoon;Yang, Young-Lyeol
    • KSBB Journal
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    • v.22 no.1
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    • pp.62-65
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    • 2007
  • New strain development strategy using kinetic models and metabolic control analysis was investigated. In this study, previously reported mathematical models describing the enzyme kinetics of intracellular threonine synthesis were modified for mutant threonine producer Escherichia coli TF5015. Using the modified models, metabolic control analysis was carried out to identify the rate limiting step by evaluating the flux control coefficient on the overall threonine synthesis flux exerted by individual enzymatic reactions. The result suggested the production of threonine could be enhanced most efficiently by increasing aspartate semialdehyde dehydrogenase (asd) activity of this strain. Amplification of asd gene in recombinant strain TF5015 (pCL-$P_{aroF}$-asd) increased the threonine production up to 23%, which is much higher than 14% obtained by amplifying aspartate kinse (thrA), other gene in threonine biosynthesis pathway.

Evaluation of Optimum Dietary Threonine Requirement by Plasma Free Threonine and Ammonia Concentrations in Surgically Modified Rainbow Trout, Oncorhynchus mykiss

  • Yun, Hyeonho;Park, Gunjun;Ok, Imho;Katya, Kumar;Heung, Silas;Bai, Sungchul C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.4
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    • pp.551-558
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    • 2015
  • This study was carried out to evaluate the dietary threonine requirement by measuring the plasma free threonine and ammonia concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A total of 70 fish (average initial weight $506{\pm}8.2g$) were randomly distributed into each of the 14 net cages (5 fish/cage). After 48 hours (h) of feed deprivation, each group was intubated at 1% body weight with one of the seven L-amino acid based diets containing graded levels of threonine (0.42%, 0.72%, 0.92%, 1.12%, 1.32%, 1.52%, or 1.82% of diet, dry matter basis). Blood samples were taken at 0, 5, and 24 h after intubation. Post-prandial plasma free threonine concentrations (PPthr) of fish 5 h after intubation with diets containing 1.32% or more threonine were significantly higher than those of fish intubated with diets containing 1.12% or less threonine (p<0.05). Post-absorptive free threonine concentrations (PAthr) after 24 h of intubation of the fish with diets containing 0.92% or more threonine were significantly higher than those of fish intubated with diets containing 0.72% or less threonine. Post-prandial plasma ammonia concentrations (PPA, 5 h after intubation) were not significantly different among fish intubated with diets containing 1.12% or less threonine, except the PPA of fish intubated with diet containing 0.42% threonine. Broken-line model analyses of PPthr, PAthr, and PPA indicated that the dietary threonine requirement of rainbow trout should be between 0.95% (2.71) and 1.07% (3.06) of diet (% of dietary protein on a dry matter basis).

Simulation Study of Dynamic Network Model for L-Threonine Biosynthesis in Escherichia coli (대장균의 동역학 네트워크 모델을 이용한 L-threonine 생합성에 관한 모사 연구)

  • Jung, Uisub;Lee, Jinwon
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.97-105
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    • 2006
  • In order to investigate the effect of inhibitors on L-threonine biosynthesis in Escherichia coli, we have constructed a metabolic network model of amino acid biosynthesis from L-aspartate to L-threonine by using available informations from literatures and databases. In the model, the effects of inhibitors on the biosynthesis of L-threonine was included as an appropriate mathematical form. For simulation study, we used initial values as L-aspartate 5 mM, ATP 5 mM, NADPH 2 mM, and observed the concentration changes of intermediate metabolites over concentration changes of respective inhibitors. As a result, we found that concentrations of intermediate metabolites were not significantly changed over concentration changes of L-lysine, L-methionine, and L-glutamate. But, there were considerable changes of intermediates over concentration changes of L-serine, L-cysteine, and L-threonine, which can be considered as essential effectors on L-threonine synthesis. Contrary, the synthesis of L-threonine seems to be not related to the amounts of L-aspartate, and inversely proportional to the accumulated amount of D,L-aspartic ${\beta}$-semialdehyde.

Studies on the Formation of L-Threonine by auxotrophic mutants of Brevibacterium flavum (Brevibacterium Flavum의 Auxotrophic Mutants에 의한 L-Threonine 생성(生成)에 관한 연구(硏究))

  • Lee, Kap-Rang;Park, Dong-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.4
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    • pp.251-261
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    • 1987
  • This study was attempted to increase the production of L-Threonine by Brevibacterium Flavum ATCC 14067, To select the strain which produce the highest threonine, mutants ere induced by N-methyl-N'-nitro-N-nitrosoguanidine treatment. The composition of media and cultural condition for its overproduction of threonine were also studied. In a threonine producer, strain B-13(Met-) was the strain producing the highest amount of threonige among methionine, lysine and isoleucine auxotrophs. The following results were obtained. 1. The wild strain and B-13(Met-) produced threonine 1.4mg/ml and 4.86mg/ml , respectively. 2. The optimum composition of medium for producing threonine by Brevibacterium Flavum B-13 was glucose 10%, ammonium sulfate 4%, potassium phosphate monobasic 0.2%, magnesium sulfate 0.05%, biotin $200{\mu}l$, thiamine $300{\mu}l$. Addition of nicotinic acid also led to increase L-threonine production. 3. In addition of organic nutrients to the fermentation medium, peptone n'ere effective and addition of methionine $100{\mu}g/ml$ produced the highest amount of L-Threonine. Aspartic acid and homoserine were also effective when these amino acid were added to the fermentstion medium. 4. Cultural conditon on threonine production by B-16 were investigated. The optimum pH was 7.0-8.0. The highest amount of threnine was produced after 4 days of cultural period.

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Isolation of the Threonine Dehydratase Gene from a Tylosin-Producing Strain of Streptomyces fradiae

  • Lee, Sang Hee;Kye Joon Lee
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.305-308
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    • 1995
  • From the plasmid library made from Sstl and San-digested genomic DNA of Streptomyces fradiae NRRL 2702, four positive clones were selected using an oligodeoxynucleotide probe from the N-terminal amino acid sequence of purified threonine dehydratase. The cloned gene for threonine dehydratase was a 2.0 kilo-base pair DNA fragment. The deduced amino acid sequence of PCR product (PCR245) was matched to that of the N-terminal part of threonine dehydratase from S. fradiae and this showed a high similarity to the threonine dehydratases of other organisms. This indicated that amino acid sequences of threonine dehydratases were highly conserved and the polypeptide product of the PCR245 was likely to be involved in the deamination of threonine.

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Partition of Amino Acid Requirements of Broilers between Maintenance and Growth. IV. Threonine and Glycine

  • Bae, S.H.;Kim, J.H.;Shin, I.S.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.3
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    • pp.381-387
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    • 1999
  • Two experiments were conducted to subdivide threonine (exp. 1) and glycine (exp. 2) requirements of broilers into maintenance and growth requirements. Purified diets containing five graded levels of threonine (exp. 1) and glycine (exp. 2) were fed to growing chicks to estimate threonine (exp. 1) and glycine (exp. 2) requirements for growth and maintenance. A model developed to divide threonine requirement for maintenance from that for growth yielded a requirement for growth of 8.946 mg/g weight gain and 0.341 mg/mg N gain; the maintenance requirement was 0.033 or 0.030 mg per unit of metabolic body size $(Wg^{0.75})$. The plateau of plasma threonine concentration occurred at 279.4 mg threonine intake/day. The total threonine requirement was 289.1 mg/day or 0.69% of the diet, 294.1 mg/day or 0.71% of the diet based on weight gain and nitrogen gain responses, respectively. These estimates were in close agreement with previous estimates of threonine requirements. From the relationship of weight gain to N gain, 5.46% of the retained protein consisted of threonine; the reported threonine content of chick muscle was 4.02%. The glycine requirement for maintenance could not be determined due to failure to obtain data allowing extrapolation to zero response. However, ADG increased slightly up to 0.56% glycine.