• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.581-581
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• 2003

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.384-384
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• 2005

• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.314-322
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• 2018

Bacillus subtilis 8 is highly efficient at degrading feather keratin. We observed integrated feather degradation over the course of 48 h in basic culture medium while studying the entire process with scanning electron microscopy. Large amounts of ammonia, sulfite, and $\text\tiny{L}$-cysteic acid were detected in the fermented liquid. In addition, four enzymes (gamma-glutamyltranspeptidase, peptidase T, serine protease, and cystathionine gamma-synthase) were identified that play an important role in this degradation pathway, all of which were verified with molecular cloning and prokaryotic expression. To the best of our knowledge, this report is the first to demonstrate that cystathionine gamma-synthase secreted by B. subtilis 8 is involved in the decomposition of feather keratin. This study provides new data characterizing the molecular mechanism of feather degradation by bacteria, as well as potential guidance for future industrial utilization of waste keratin.

• 약학회지
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• 제53권2호
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• pp.74-77
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• 2009

Food deprivation decreases hepatic glutathione (GSH) levels, which is ascribed to alterations in availability of hepatic cysteine, a rate limiting factor for the GSH synthesis. The present study examines the effects of food deprivation on hepatic metabolism of sulfur amino acid in male rats. In rats fasted for 24 or 48 hours, hepatic GSH levels were decreased from $6.70{\pm}0.16{\mu}mol/g$ liver to $4.02{\pm}0.20$ or $4.06{\pm}0.07{\mu}mol/g$ liver, respectively. Hepatic S-adenosylmethionine levels were also decreased in fasted rats, but S-adenosylhomocysteine levels were increased. Hepatic methionine levels were not changed by food deprivation for 48 hours. On the other hand, hepatic cysteine or taurine levels were increased from $106.2{\pm}4.1$ to $130.0{\pm}2.7$ nmol/g liver or from $2.45{\pm}0.43$ to $5.07{\pm}0.78{\mu}mol/g$ liver, respectively, in 48-hour fasted rats. Activity of cystathionine beta-synthase catalyzed homocysteine to cystathionine, was markedly decreased, but activity of betaine homocysteine methyltransferase was increased in fasted rats, indicating that methylation of homocysteine to methionine is activated. Also activity of cysteine dioxygenase, involved in taurine synthesis, was increased. These results suggested that hepatic methionine levels were maintained in rats fasted for 48 hours through increase in homocysteine methylation, and hepatic GSH may serve as a cysteine supplier reservoir in fasting state.

• KSBB Journal
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• 제23권5호
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• pp.425-430
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• 2008

생체에서의 황화수소는 의약 분야와 발효 산업에 있어서 중요한 역할을 하고 있다. 본 연구에서는 Saccharomyces cerevisiae을 이용하여 기질인 L-cysteine과 $\beta$-mercaptoethanol로부터 $\beta$-replacement 반응에 인한 황화수소를 발생시킬 수 있는 효소를 간편하고 신속하게 동정하는 방법의 확립을 목적으로 하였다. 효소에 의해 발생된 황화수소와 Pb-acetate의 반응으로 생성된 Pb-S를 gel상에서 간편하게 확인한 후, 확인된 단백질들을 이온교환컬럼를 수행한 후 gel에서 추출하는 방법으로 MALDI-TOFMS의 시료를 간단히 얻을 수 있었다. PMF 방법과 MS/MS ion search 분석을 통해 간편하게 효모에서 황화수소를 형성할 가능성이 있는 세가지 단백질의 동정에 성공하였다. 이 세 가지 단백질은 CYS3, CYS4, MET17 유전자의 단백질로서 cystathionine $\gamma$-lyase, CBS, OASS 임이 밝혀졌다. 그리고 이 세 단백질들은 L-cysteine과 $\beta$-mercaptoethanol의 존재하에서 황화수소를 실제로 생성함을 확인하였다. 본 연구에서 표적의 물질을 생산하는 단백질들을 젤상에서의 활성측정과 MALDI-TOF MS를 이용하여 간단히 그리고 정확하게 동정하는 방법을 확립하였다.

• 대한유전성대사질환학회지
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• 제15권2호
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• pp.78-86
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• 2015

목적: 호모시스틴뇨증은 cystathionine ${\beta}$-synthase의 결핍으로 유발되는 질환이다. 본원에서 치료받고 있는 총 6명 환자들의 임상양상과 유전자 분석을 시행하였다. 방법: 1992년 1월부터 2015년 3월까지 본원에서 전형적 호모시스틴뇨증으로 진단되어 치료 중인 총 6명의 환자들을 대상으로 후향적으로 임상적, 생화학적, 유전자적 분석을 하였다. 결과: 총 6명의 환자가 본 연구에 포함되었으며 이 중 3명은 영아기에 발견되었고, 나머지 3명은 7세 이후에 진단되었다. 영아기에 진단되어 치료를 받은 환자들은 정상 인지기능을 보였고, 7세 이후에 진단된 환자들은 다양한 정도의 지적 장애가 있었다. 골감소증이 6명 중 2명의 환자에서 발견되었다. 혈전 색전증이나 경련, 수정체 탈구 같은 주요한 합병증은 7세 이후에 진단된 군에만 있었다. 유전자 검사에서 모두 cystathionine ${\beta}$-synthase 유전자의 돌연변이를 확인하였다. 본 연구에 포함된 6명은 현재 메티오닌 제한 식이를 하면서 엽산과 베타인, 비타민 B6, 메틸코발라민을 복용 중이다. 결론: 6명의 전형적 호모시스틴뇨증 환자들을 길게는 23년 동안 추적 관찰하였다. 신생아 대사 이상 검사에서 발견된 환자들은 합병증 없이 성장하고 있으나, 7세 이후에 진단된 환자들은 100% 수정체 탈구가 있었고 각종 합병증이 동반되었다. 따라서 전형적 호모시스틴뇨증의 예후에는 무엇보다 조기 진단이 중요하며 이는 현재 국내에서 시행되는 신생아 대사 이상 검사의 중요성을 설명하는 것이라 하겠다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.166-166
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• 2001

Effects of propargylglycine (PPG) treatment on the hepatic glutathione (GSH) synthesis were examined in adult male ICR mice. Administration of PPG (200 mole/kg, ip) to mice resulted in a complete inhibition of the hepatic cystathionine ${\gamma}$ -lyase (C ${\gamma}$ L) activity measured in cytosol fraction for 40 hr after the treatment. A single injection of PPG rapidly reduced the hepatic GSH levels, which appeared to be sustained at least for 40 hr.(omitted)

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.1010-1017
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• 2007

Two alternative pathways for methionine biosynthesis are known in Corynebacterium glutamicum: one involving transsulfuration (mediated by metB and metC) and the other involving direct sulfhydrylation (mediated by metY). In this study, MetB (cystathionine ${\gamma}-synthase$) and MetY (O-acetylhomoserine sulfhydrylase) from C. glutamicum were purified to homogeneity and the biochemical parameters were compared to assess the functional and evolutionary importance of each pathway. The molecular masses of the native MetB and MetY proteins were measured to be approximately 170 and 280 kDa, respectively, showing that MetB was a homotetramer of 40-kDa subunits and MetY was a homohexamer of 45-kDa subunits. The $K_m$ values for the O-acetylhomoserine catalysis effected by MetB and MetY were 3.9 and 6.4 mM, and the maximum catalysis rates were $7.4\;(k_{cat}=21\;S^{-1})\;and\;6.0\;(k_{cat}=28\;S^{-1})\;{\mu}mol\;mg^{-1}\;min^{-1}$, respectively. This suggests that both MetB and MetY can be comparably active in vivo. Nevertheless, the $K_m$ value for sulfide ions by MetY was 8.6mM, which was too high, considering the physiological condition. Moreover, MetB was active at a broad range of temperatures $(30\;and\;65^{\circ}C)$ and pH (6.5 and 10.0), as compared with MetY, which was active in a range from 30 to $45^{\circ}C$ and at pH values from 7.0 to 8.5. In addition, MetY was inhibited by methionine, but MetB was not. These biochemical data may provide insight on the role of the parallel pathways of methionine biosynthesis in C. glutamicum with regard to cell physiology and evolution.

• The Korean Journal of Physiology and Pharmacology
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• 제25권3호
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• pp.239-249
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• 2021

The present study explored the therapeutic potential of hydrogen sulfide (H2S) in restoring aging-induced loss of cardioprotective effect of remote ischemic preconditioning (RIPC) along with the involvement of signaling pathways. The left hind limb was subjected to four short cycles of ischemia and reperfusion (IR) in young and aged male rats to induce RIPC. The hearts were subjected to IR injury on the Langendorff apparatus after 24 h of RIPC. The measurement of lactate dehydrogenase, creatine kinase and cardiac troponin served to assess the myocardial injury. The levels of H2S, cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), nuclear factor erythroid 2-related factor 2 (Nrf2), and hypoxia-inducible factor (HIF-1α) were also measured. There was a decrease in cardioprotection in RIPC-subjected old rats in comparison to young rats along with a reduction in the myocardial levels of H2S, CBS, CSE, HIF-1α, and nuclear: cytoplasmic Nrf2 ratio. Supplementation with sodium hydrogen sulfide (NaHS, an H2S donor) and l-cysteine (H2S precursor) restored the cardioprotective actions of RIPC in old hearts. It increased the levels of H2S, HIF-1α, and Nrf2 ratio without affecting CBS and CSE. YC-1 (HIF-1α antagonist) abolished the effects of NaHS and l-cysteine in RIPC-subjected old rats by decreasing the Nrf2 ratio and HIF-1α levels, without altering H2S. The late phase of cardioprotection of RIPC involves an increase in the activity of H2S biosynthetic enzymes, which increases the levels of H2S to upregulate HIF-1α and Nrf2. H2S has the potential to restore aging-induced loss of cardioprotective effects of RIPC by upregulating HIF-1α/Nrf2 signaling.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.114-126
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• 2019

This paper introduces three ways to determine host-guest complexation of cucurbit[7]uril (CB[7]) with homocysteine (Hcy). After preincubating Hcy and cysteine (Cys) with CB[7], Ellman's reagent (DTNB) was used to detect Hcy and Cys. Only Cys reacted with DTNB and Hcy gave a retarded color change. This suggests that the -SH group of Hcy is buried inside CB[7]. Human cystathionine ${\gamma}-lyase$ (hCGL) decreased the level of Hcy degradation after preincubating Hcy and CB[7]. These results suggest that the amount of free Hcy available was decreased by the formation of a Hcy-CB[7] complex. The immunological signal of anti-Hcy monoclonal antibody was decreased significantly by preincubating CB[7] with Hcy. The ELISA results also show that ethanethiol group ($-CH_2CH_2SH$) of Hcy, which is an epitope of anti-Hcy monoclonal antibody, was blocked by the cavity in CB[7]. Overall, CB[7] can act as a host by binding selectively with Hcy, but not Cys. The calculated half-complexation formation concentration of CB[7] was 58.2 nmol using Ellman's protocol, 97.9 nmol using hCGL assay and 87.7 nmol using monoclonal antibody. The differing binding abilities of Hcy and Cys towards the CB[7] host may offer a simple and useful method for determining the Hcy concentration in plasma or serum.