• Title/Summary/Keyword: L-Met

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Direct Conversion of L-Selenomethionine into Methylselenol by Human Cystathionine ${\gamma}$-Lyase (인간 Cystathionine ${\gamma}$-Lyase에 의한 Selenomethionine의 Methylselenol로의 직접분해)

  • Cho, Hyun-Nam;Jhee, Kwang-Hwan
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.11-17
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    • 2014
  • Selenium is an essential trace element for mammals, but it is very toxic. Therefore, the control of selenium concentrations should be precisely and effectively monitored. Selenium is naturally obtained through foods and seleno-L-methionine (LSeMet) is a major form of selenium. It has been reported that L-SeMet is only converted into Se-adenosyl-L-SeMet. However, a recent study suggested that L-SeMet was directly metabolized into methylselenol ($CH_3SeH$) in mouse liver extract by the reaction of cystathionine ${\gamma}$-lyase (CGL). The canonical reaction of CGL was known to catalyze the cleavage of L-cystathionine to L-cysteine, ${\alpha}$-ketobutyrate and $NH_3$. In the present study, we found that L-SeMet could be directly converted to $CH_3SeH$ using purified homogenous human CGL instead of mouse liver cytosol. Authentic $CH_3SeH$ was prepared by reduction of dimethyldiselenide with sodium tetrahydroborate. The gaseous product of the enzymatic reaction with L-SeMet was analyzed by GC/MS spectrometry. The GC/MS data was identical to that of authentic dinitrophenyl selenoether. We also analyzed the kinetic parameters for the formation of $CH_3SeH$ from L-SeMet by human and mouse CGL. These results suggest that human CGL is a critical enzyme which is responsible for L-SeMet metabolism.

Enhanced supply of methionine regulates protein synthesis in bovine mammary epithelial cells under hyperthermia condition

  • Zhou, Jia;Yue, Shuangming;Xue, Benchu;Wang, Zhisheng;Wang, Lizhi;Peng, Quanhui;Xue, Bai
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1126-1141
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    • 2021
  • Recent evidence has shown that methionine (Met) supplementation can improve milk protein synthesis under hyperthermia (which reduces milk production). To explore the mechanism by which milk protein synthesis is affected by Met supplementation under hyperthermia, mammary alveolar (MAC-T) cells were incubated at a hyperthermic temperature of 42℃ for 6 h in media with different concentrations of Met. While the control group (CON) contained a normal amino acid concentration profile (60 ㎍/mL of Met), the three treatment groups were supplemented with Met at concentrations of 10 ㎍/mL (MET70, 70 ㎍/mL of Met), 20 ㎍/mL (MET80, 80 ㎍/mL of Met), and 30 ㎍/mL (MET90,90 ㎍/mL of Met). Our results show that additional Met supplementation increases the mRNA and protein levels of BCL2 (B-cell lymphoma-2, an anti-apoptosis agent), and decreases the mRNA and protein levels of BAX (Bcl-2-associated X protein, a pro-apoptosis agent), especially at an additional supplementary concentration of 20 ㎍/mL (group Met80). Supplementation with higher concentrations of Met decreased the mRNA levels of Caspase-3 and Caspase-9, and increased protein levels of heat shock protein (HSP70). The total protein levels of the mechanistic target of rapamycin (mTOR) and the mTOR signalling pathway-related proteins, AKT, ribosomal protein S6 kinase B1 (RPS6KB1), and ribosomal protein S6 (RPS6), increased with increasing Met supplementation, and peaked at 80 ㎍/mL Met (group Met80). In addition, we also found that additional Met supplementation upregulated the gene expression of αS1-casein (CSN1S1), β-casein (CSN2), and the amino acid transporter genes SLC38A2, SLC38A3 which are known to be mTOR targets. Additional Met supplementation, however, had no effect on the gene expression of κ-casein (CSN3) and solute carrier family 34 member 2 (SLC34A2). Our results suggest that additional Met supplementation with 20 ㎍/mL may promote the synthesis of milk proteins in bovine mammary epithelial cells under hyperthermia by inhibiting apoptosis, activating the AKT-mTOR-RPS6KB1 signalling pathway, and regulating the entry of amino acids into these cells.

Effect of L- or DL-methionine Supplementation on Nitrogen Retention, Serum Amino Acid Concentrations and Blood Metabolites Profile in Starter Pigs

  • Tian, Q.Y.;Zeng, Z.K.;Zhang, Y.X.;Long, S.F.;Piao, X.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.689-694
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    • 2016
  • The objective of the current study was to evaluate the effect of supplementation of either L-methionine (L-Met) or DL-methionine (DL-Met) to diets of starter pigs on nitrogen (N) balance, metabolism, and serum amino acid profile. Eighteen crossbred ($Duroc{\times}Landrace{\times}Yorkshire$) barrows weighing $15.45{\pm}0.88kg$ were randomly allotted to 1 of 3 diets with 6 pigs per treatment. The diets included a basal diet (Met-deficient diet) containing 0.24% standardized ileal digestibility Met with all other essential nutrients meeting the pig's requirements. The other two diets were produced by supplementing the basal diet with 0.12% DL-Met or L-Met. The experiment lasted for 18 days, consisting of a 13-day adaptation period to the diets followed by a 5-day experimental period. Pigs were fed ad libitum and free access to water throughout the experiment. Results showed that the supplementation of either L-Met or DL-Met improved N retention, and serum methionine concentration, and decreased N excretion compared with basal diet (p<0.01). The N retention of pigs fed diets supplemented with the same inclusion levels of DL-Met or L-Met were not different (p>0.05). In conclusion, on equimolar basis DL-Met and L-Met are equally bioavailable as Met sources for starter pigs.

Differential Response of Etiolated Pea Seedlings to Inoculation with Rhizobacteria Capable of Utilizing 1-Aminocydopropane-1-Carboxylate or L-Methionine

  • Shaharoona, Baby;Arshad, Muhammad;Khalid, Azeem
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.15-20
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    • 2007
  • The majority of soil microorganisms can derive ethylene from L-methionine (L-MET), while some rhizobacteria can hydrolyze 1-aminocyclopropane-1-carboxylate (ACC) due to their ACC-deaminase activity. In this study, three strains having either ACC-deaminase activity (Pseudomonas putida biotype A, $A_7$), or the ability to produce ethylene from L-MET (Acinetobacter calcoaceticus, $M_9$) or both (Pseudomonas fluorescens, $AM_3$) were used for inoculation. The highly ethylene specific bioassay of a classical 'triple' response in pea seedlings was used to investigate the effect of the inoculation with the rhizobacteria in the presence of 10 mM ACC or L-MET. The exogenous application of ACC had a concentration-dependent effect on the etiolated pea seedlings in creating the classical 'triple' response. The inoculation with P. putida diluted the effect of ACC, which was most likely due to its ACC-deaminase activity. Similarly, the application of $Co^{2+}$ reduced the ACC-imposed effect on etiolated pea seedlings. In contrast, the inoculation of A. calcoaceticus or P. fluorescens in the presence of L-MET caused a stronger classical 'triple' response in etiolated pea seedlings; most likely by producing ethylene from L-MET. This is the first study, to our knowledge, reporting on the comparative effect of rhizobacteria capable of utilizing ACC vs L-MET on etiolated pea seedlings.

Associations between Anemia and Glomerular Filtration Rate and Albuminuria in Korean Adults by Metabolic Syndrome Status: Analysis of KNHNES V-3 Data (대한민국 성인의 대사증후군 유무에 따른 빈혈과 사구체 여과율 및 알부민뇨의 연관성: 국민건강영양조사 V-3 분석)

  • Hyun YOON
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.2
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    • pp.125-134
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    • 2024
  • The present study was conducted to explore relationships between anemia and estimated glomerular filtration rate (eGFR) and urine microalbumin/creatinine ratio (uACR) in Korean adults with or without metabolic syndrome (MetS). The data of 4,943 adults aged ≥20 years who participated in KNHNES V-3 (2012) were analyzed. In the non-MetS group, the odds ratio (OR) for anemia of those with a decreased eGFR {eGFR<60 mL/min/1.73 m2, 3.85 (95% confidence interval [CI], 2.03~7.30)} was significant as was the OR of those with decreased eGFR plus elevated uACR (eGFR<60 mL/min/1.73 m2 and uACR≥30 mg/g, 5.81 [95% CI, 2.60~13.02]). In the MetS group, ORs for anemia for those with an elevated uACR (2.18 [95% CI, 1.11~4.27]), a decreased eGFR (3.74 [95% CI, 1.11~12.55]), or a decreased eGFR plus an elevated uACR (16.79 [95% CI, 5.93~47.57]) were significant. In conclusion, in non-MetS, anemia was associated with a low eGFR, whereas in MetS, anemia was associated with a low eGFR and an elevated uACR. In addition, the OR for anemia was greatly increased when eGFR was diminished and uACR was elevated regardless of MetS and MetS status.

Improving the Productivity of Single-Chain Fv Antibody Against c-Met by Rearranging the Order of its Variable Domains

  • Kim, Yu-Jin;Neelamegam, Rameshkumar;Heo, Mi-Ae;Edwardraja, Selvakumar;Paik, Hyun-Jong;Lee, Sun-Gu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1186-1190
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    • 2008
  • Single-chain Fv (scFv) antibody against c-Met is expected to be employed in clinical treatment or imaging of cancer cells owing to the important biological roles of c-Met in the proliferation of malignancies. Here, we show that the productivity of scFv against c-Met in Escherichia coli is significantly influenced by the orientation of its variable domains. We generated anti-c-Met scFv antibodies with two different domain orders (i.e., $V_L$-linker-$V_H$ and $V_H$-linker-$V_L$), expressed them in the cytoplasm of E. coli trx/gor deleted mutant, and compared their specific activities as well as their productivities. Productivity of total and functional anti-c-Met scFv with $V_H/V_L$ orientation was more than five times higher than that with $V_L/V_H$ format. Coexpression of DsbC enhanced the yield of soluble amounts of anti-c-Met scFv protein for both constructs. The purified scFv antibodies of the two different formats exhibited almost the same antigen-binding activities. We also compared the productivities and specific activities of anti-c-Met diabodies with $V_H/V_L$ or $V_L/V_H$ formats and obtained similar results to the case of scFv antibodies.

Distribution of S-Adenosylmethionine Synthetase in the Pancreatic Tissues of Various Animals and Changes of S-Adenosylmethionine Synthetase Activities and S-Adenosylmethionine in the Developing Rat Organs (췌조직과 성장 발육에 따른 흰쥐 조직내 S-Adenosylmethionine Synthetase 활성도 및 S-Adenosyl-L-methionine의 분포)

  • Park, Seung-Hee;Yu, Tae-Moo;Hong, Sung-Youl;Lee, Hyang-Woo
    • YAKHAK HOEJI
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    • v.38 no.4
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    • pp.430-439
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    • 1994
  • S-Adenosyl-L-methionine synthetase (ATP: methionine S-Adenosyltransferase, EC 2.5.1.6; AdoMet synthetase) catalyzes the biosynthesis of S-Adenosyl-L-methionine(AdoMet) from methionine in the presence of ATP. To elucidate the role of transmethylation reaction in the pancreatic tissues, we examined AdoMet synthetase and isozyme activities, and AdoMet contents in the various tissues. The activities of AdoMet synthetase marked the highest in the kidney, and the lowest in the testis among the various tissues of rat. Considerable amounts of AdoMet synthetase activities were detected in the pancreatic tissues of various animals except for those of frog. The level of ${\alpha}$ and ${\gamma}$ isozyme activities were present in the pancreatic tissues of various animals, while ${\beta}$ isozyme activities were detected as trace. AdoMet synthetase activities of rat brain, liver, testis were decreased with growth. In the rat pancreatic tissues, AdoMet synthetase activities were increased during 16 days after birth and then decreased between 16 and 47 days of age. Levels of AdoMet contents of rat brain and testis were decreased with growth. However, AdoMet contents of rat pancreas were decreased until 26 days of age, and then increased thereafter. AdoMet synthetase isozyme patterns did not vary with growth in the pancreas and testis. But, in the liver, ${\beta}$ form is strikingly increased with growth.

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Growth Characteristics of Acidithiobacillus thiooxidans in Different Sulfur Concentrations (황 농도에 따른 Acidithiobacillus thiooxidans의 생장 특성)

  • Lee, Eun-Young;Cho, Kyung-Suk;Ryu, Hee-Wook
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.338-341
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    • 2006
  • The growth characteristics of sulfur-oxidizing bacteria, Acidithiobacillus thiooxidans AZ11, MET, and TAS were investigated in mineral salt media supplemented with elemental sulfur of 1$\sim$50 g $L^{-1}$. The sulfur oxidation rates of A. thiooxidans. MET and TAS increased highly with increasing sulfur concentration up to 10 g L$^{-1}$, but the rates increased slowly in sulfur concentration over 10 g L$^{-1}$. A. thiooxidans AZ11 showed the parallel increase of sulfur oxidation rate until sulfur concentration increased up to 40 g L$^{-1}$. The maximum sulfur oxidation rates (V$_{max}$) of AZl1, MET and TAS were 1.88, 1.38 and 0.43 g S L$^{-1}$ d$^{-1}$, respectively. The maximum specific growth rates (${\mu}_{max}$) of AZ11, MET, and TAS were 0.33 d$^{-1}$, 0.30 d$^{-1}$ and 0.45 d$^{-1}$, respectively. Although MET and TAS couldn't grow at sulfate concentration of 40 g L$^{-1}$, AZ11 could grow in the presence of 58 g L$^{-1}$ sulfate, the final oxidation product of elemental sulfur.

Cloning and Functional Analysis of Gene Coding for S-Adenosyl-L-Methionine Synthetase from Streptomyces natalensis (Streptomyces natalensis로부터 S-adenosyl-L-methionine synthetase 유전자의 클로닝 및 기능분석)

  • Yoo, Dong-Min;Hwang, Yong-Il;Choi, Sun-Uk
    • Journal of Life Science
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    • v.21 no.1
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    • pp.96-101
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    • 2011
  • S-Adenosyl-L-methionine synthtase (SAM-s) catalyzes the biosynthesis of SAM from ATP and L-methionine. SAM plays important roles in the primary and secondary metabolism of cells. A metK encoding a SAM-s was searched from Streptomyces natalensis producing natamycin, a predominantly a strong antifungal agent, inhibiting the growth of both yeasts and molds and preventing the formation of aflatoxin in filamentous fungi. To obtain the metK of S. natalensis, PCR using primers designed from the two highly conserved regions for metK genes of Streptomyces strains was carried out, and an intact 1.2-kb metK gene of S. natalensis was cloned by genomic Southern hybridization with PCR product as a probe. To identify the function of the cloned metK gene, it was inserted into pSET152ET for its high expression in the Streptomyces strain, and then introduced into S. lividans TK24 as a host by transconjugation using E. coli ET12567(pUZ8002). The high expression of metK in S. lividans TK24 induced actinorhodin production on R5 solid medium, and its amount in R4 liquid medium was 10-fold higher than that by exconjugant including only pSET152ET.