• 한국미생물·생명공학회지
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• 제42권1호
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• pp.11-17
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• 2014

Selenium(Se)은 사람에게 필수성분이지만 독성이 강한 물질이다. 따라서 생체에서 Se(셀레늄)의 농도조절은 정확하고 효율적으로 이루어져야 한다. 사람은 Se을 음식의 형태로 섭취해야 하고 주로 seleno-L-methionine (L-SeMet)의 형태로 섭취한다. 섭취된 L-SeMet은 methionine 대사와 동일한 효소를 이용하여 Se-adenosyl-L-SeMet으로 대사된다고 알려져 있었다. 그러나 최근 쥐의 간 추출물의 실험에서, L-SeMet이 cystathionine ${\gamma}$-lyase (mouse CGL)의 작용으로 methylselenol ($CH_3SeH$)로 직접적으로 대사된다는 보고가 있었다. CGL은 원래 cystathionine을 L-cysteine과 ${\alpha}$-ketobutyrate, 그리고 $NH_3$로 분해하는 효소로 알려져 있었다. 따라서 본 연구에서는 쥐의 간 추출물 대신에 인간의 CGL을 분리 정제하여 L-SeMet에서 methylselenol의 형성을 확인하고자 하였다. Methylselenol의 표준시료는 dimethyldiselenide를 sodium tetrahydroborate로 환원시켜 준비하였다. 그리고 L-SeMet을 기질로 사용한 효소 반응액 중에서 가스상의 생성물은 GC/MS 스펙트럼으로 분석하였다. 분석 결과 methylselenol의 유도체인 dinitrophenyl selenoether와 일치하였다. 또한 인간 CGL이 L-SeMet에서 methylselenol을 형성하는 반응의 kinetic parameter를 mouse CGL과 비교 분석하였다. 결과적으로 human CGL은 섭취된 L-SeMet의 대사를 책임지고 있는 중요한 효소이다.

• Journal of Animal Science and Technology
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• 제63권5호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제29권5호
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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.

• Journal of Microbiology
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• 제45권1호
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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.

• 대한임상검사과학회지
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• 제56권2호
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• pp.125-134
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• 2024

본 연구는 대한민국 성인을 대상으로 대사증후군(metabolic syndrome, MetS) 유·무에 따른 빈혈과 추정 사구체여과율(estimated glomerular filtration rate, eGFR) 및 요 미세 알부민/크레아티닌 비율(urine microalbumin/creatinine ratio, uACR)의 관련성을 평가하기 위하여 2012년 국민건강 영양조사(KNHNES V-3) 자료를 활용하여 20세 이상 성인 4,943명을 대상으로 데이터를 분석하였다. 본 연구에서 몇 가지 중요한 발견이 있었다. 첫째, 비 MetS 그룹에서는 정상군(eGFR≥60 mL/min/1.73 m² 및 uACR<30 mg/g)의 빈혈(남성, 헤모글로빈[hemoglobin, Hb]<13 g/dL; 여성, Hb<12 g/dL)의 발생률에 비하여 감소된 eGFR 그룹(eGFR<60 mL/min/1.73 m²; odds ratio [OR], 3.65; 95% confidence interval [CI], 1.90~7.00) 및 감소된 eGFR+증가된 uACR 그룹(eGFR<60 mL/min/1.73 m² 및 uACR≥30 mg/g, OR, 6.00; 95% CI, 2.61~13.80)의 빈혈 발생률이 높았다. 둘째, MetS 그룹에서는 정상군에 비하여 증가된 uACR 그룹(OR, 2.18; 95% CI, 1.11~4.27), 감소된 eGFR 그룹(OR, 3.73; 95% CI, 1.09~12.75) 및 감소된 eGFR+증가된 uACR 그룹(OR, 18.17; 95% CI, 6.16~53.63)의 빈혈 발생률이 높았다. 결론적으로, 비 MetS 그룹에서는 빈혈은 eGFR의 감소와 관련이 있었고, MetS 그룹에서는 빈혈은 eGFR 감소 및 uACR 증가와 관련이 있었다. 추가적으로, 비 MetS 그룹과 MetS 그룹 모두에서 eGFR의 감소 및 uACR의 증가가 동시에 나타날 때 빈혈의 발생률이 크게 증가하였다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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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.

• 수산해양기술연구
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• 제27권4호
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• pp.232-237
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• 1991

통발어구의 미끼를 개선하기 위한 기초실험으로 16종의 천연미끼와 아미노산 및 핵산 관련물질을 사용하여 현장에서 어획실험을 행한 결과는 다음과 같다. 1. 주된 어획 생물은 붕장어, 왜볼락, 망둑, 게, 골뱅이 및 불가사리 등이었다. 2. 어획 생물별로 유인성활이 강한 천연미끼는 아래와 같다. 붕장어: 멸치, 정어리, 참갯지렁이. 망둑: 멸치, 정어리, 참갯지렁이, 명태, 크릴. 왜볼락: 줄새우, 갯가재. 게: 꽃새우, 크릴, 오징어육, 멸치, 명태, 바위갯지렁이. 골뱅이: 멸치, 정어리, 털게, 꽃새우, 개조개. 불가사리: 정어리, 우럭조개, 개조개, 오징어육. 3. 아미노산과 핵산관련물질은 천연미끼보다 유인활성이 낮았지만 비교적 어획을 많이 보인 물질은 다음과 같다. 붕장어: Taurine, L-Glu, L-Gly, L-Met, IMP. 망둑: L-Glu, L-Met. 왜볼락: L-Met. 게: Taurine. Hx. 골뱅이: L-Gly, L-Met. 불가사리: IMP.

• 약학회지
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• 제38권4호
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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.

• 한국미생물·생명공학회지
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• 제34권4호
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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.

• 생명과학회지
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• 제21권1호
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• pp.96-101
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• 2011

ATP와 L-methionine으로부터 SAM synthetase (MetK)에 의해 생합성 되는 S-adenosylmethionine (SAM)은 세포내 메틸화에 필요한 메틸기를 제공하는 중심적인 공급체의 역할을 할뿐만 아니라 방선균에서는 일차 및 이차대사산물의 생산 조절에 관여하고 있다는 사실이 밝혀졌다. 이에 논 연구에서는 산업적으로 매우 중요한 항진균성 항생물질인 natamycin을 생산하는 S. natalensis로부터 SAM synthetase 코드하는 metK 유전자를 클로닝하고 동정하였다. S. natalensis에서 클로닝된 metK는 1,209 bp의 염기를 가진 유전자로써 아미노산서열에서 S. pristinaespiralis ATCC 25486과 S. peucetius ATCC 27952의 MetK와 96%, S. violaceusniger Tu 4113과 95% 일치하는 매우 높은 상동성을 보였다. 또 pSET152ET 벡터를 이용해 구축한 metK 고발현용 재조합 플라스미드 pCD1를 S. lividans TK24의 genomic DNA에 도입하여 actinorhodin 생산 유도를 시도해 본 결과 R5 고체배지에서 pCD1이 도입되지 않은 균주에서는 actinorhodin 생산을 전혀 확인할 수 없었지만 pCD1이 도입된 형질전환체에서는 actinorhodin 생산이 강하게 유도되었으며 R4 액체배지에서는 actinorhodin 생산량이 10배 증가되었다. 따라서 본 연구를 통해 클로닝된 S. natalensis 유래 metK 유전자는 방선균에서 이차대사산물의 생산을 유도할 수 있음을 확인할 수 있었다.