• Journal of Microbiology
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• 제35권2호
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• pp.97-102
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• 1997

Cysteine desulfhydrase (EC 4.4.1.1.) was purified from the culture supernatant of Streptomyces albidoflavus SMF301 by hydroxyapatite, gel filtration and Resource Q ion-exchange chromatography with a purification fold of six identical subunits. The enzyme was stabilized by dithiothreitol and pyridoxal 5'-phosphate during the purification procedures. The optimum pH and temperature were pH 8.6 and 35$^{\circ}C$, respectively. The N-terminal amino acid sequence was identified as A-P-L-P-T-A-D-V-R-S-D-P-G-Y-R-E-W-L-G-E-A-V. The purified cystein desulfhydrase had a high substrate specificity toward cysteine, and exhibited no cystahionine $\gamma$-lyase activity. The $K_m$ value for cysteine was determined to be 0.37 mM.

• Journal of Nutrition and Health
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• 제29권7호
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• pp.729-737
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• 1996

Activieties of hepatic cysteine desulfhydration was assessed in cats fed one of the following diets for 5 weeks : 20% protein, 0% taurine diet(LPOT) ; 20% protein, 0.15% taurine diet (LPNT) ; 60% protein, 0% taurine diet(HPOT) ; and 60% protein, 0.15% taurine diet(HPNT). Cats fed LPOT and HPOT had been maintained on a taurine-free diet for 6 weeks prior to the experiment in order to deplete body taurine. Activities of cysteine desulfhydration were determined by measuring the production of H235S from 35S-cysteine in the presence and absence of $\alpha$-ketoglutarate ($\alpha$-KG) in the incubation medium. The direct pathway via cysteine desulfhydrase appears to account for the major route of cysteine desulfhydration in the cat liver since the values obtained in the absence of $\alpha$-KG were between 81 and 88% of those obtained in the presence of $\alpha$-KG. Mean$\pm$SEM of the hepatic total desulfhydration activities(umol H2S.min-1.kg body wt-1)in cats fed LPOT, LPNT, HPOT and HPNT were 117$\pm$6, 135$\pm$10, 137$\pm$10, and 190$\pm$9, respectively. The capacity of hepatic cysteine desulfhydration (UA/kg body wt) was positively cerrelated not only with the dietary concentration of taurine but also with the concentration of protein.

• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.159-164
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• 1994

Myceliuml growth and spore formation of Streptomyces albidoflavus SMF301 in submerged culture were compared with the metabolism of cysteine. Cysteine added to the culture was metabolized by cysteine desulfhydrase (EC 4.4.1.1.) to produce ammonium ions, hydrogen sulfide, and pyruvate. The redox potential of the culture broth was lowered immediately as the result of the metabolism of cysteine, which caused a lag period of mycelium growth. However enhanced activities of pyruvate dehydrogenase and a-ketoglutarate dehydrogenase were confirmed in the culture containing cysteine, indicating that pyruvate was utilized to support further mycelium growth.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.50-53
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• 1991

The reaction steps involved in the bioconversion of a chemically synthesized precursor, $D,L-2-amino-{\Delta}^2-thiazoline-4-carboxylic$ acid (D,L-ATC), to L-cysteine and the properties of the involved enzymes were investigated. It was found that the conversion consisted of two steps, i. e., D,L-ATC to S-carbamyl-L-cysteine (S-C-L-cysteine) and S-C-L-cysteine to L-cysteine, and the S-C-L-cysteine was an intermediate between them. While the enzymes involved in the reactions were induced by the addition of D,L-ATC as an inducer, S-C-L-cysteine induced only the enzyme involved in the latter step. The conversion of S-C-L-cysteine to L-cysteine could be also carried out in the presence of hydroxylamine and its rate was much faster than that by the corresponding enzyme. On the other hand, L-cysteine (or L-cystine) was decomposed to evolve $H_2S$ by the enzyme considered to be a kind of desulfhydrase. However, hydroxylamine was a perfect inhibitor for this enzyme.

• 한국미생물·생명공학회지
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• 제2권1호
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• pp.45-50
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• 1974

1. L-cysteine을 pyruvate, sulfide 및 ammonia로 분해하는 반응을 촉매하는 효소인 cysteinedesulfhydrase의 촉매기능에 대해 연구해온 바 Aerobacter aerogenes로부터 유도생산한 cysteinedesulfhydrase를 사용하여 분해반응의 역반응에 의해 pyruvate, ammonia 및 sulfide로부터 cysteine의 유도체인 S-methyl-L-cysteine 및 S-ethyl-L-cysteine을 합성하였다. 2. 합성반응에 있어서 S-methyl-L-cysteine 및 S-ethyl-L-cysteine의 생성량은 반응시간과 효소량에 비예적인 관계를 나타내었고 반응의 최적 pH는 10.0이었다. 3. 효소적 합성법에 의해 생산된 S-methyl-L-cysteine 및 S-ethyl-L-cysteine을 반응액으로 부터 단리, 결정화해서 이들 합성산물에 대한 Ion exchange chromatogram, NMR spectrum, element analysis, molecular weight 및 melting point 측정 등의 분석시험을 행한 바 이들 화합물에 대한 이론치와 잘 일치되는 결과를 얻었다.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.681-686
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• 1992

D,L-ATC의 L-cysteine으로의 생물학적 전환에 있어 균체를 이용할 때, 계면활성제의 영향, 효소의 안정성 및 연속생산공정중의 고정화 균체 반응기의 안정성에 대하여 분석하였다. 계면활성제의 첨가없이 균체만을 이용할 때 반응은 매우 미미하게 이루어졌으나 SDS와 Triton X-100을 첨가할 때 cell-free 조효소액을 이용하는 경우와 비슷한 정도의 결과가 얻어졌다. 효소 활성은 $30^{\circ}C$에서 7시간 저장후 50 저하되었으며 질소가스하의 혐기적인 조건에서는 활성저하가 거의 일어나지 않았다. 이와 같은 효소의 불활성화는 효소에 대한 산소의 작용으로 사료되었다. 그러나, alginate로 고정화한 균체를 이용한 연속반응공정에서 혐기적으로 조건하에서 150시간 내에 대부분의 활성을 잃어버렸으며, L-cysteine 저해제인 hydroxylamine을 첨가할 때 효소활성이 급속히 감소되었다.