• Journal of Microbiology
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• 제33권3호
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• pp.239-243
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• 1995

Deinococcus radiophilus, an UV resistant bacterium seemed to contain three issoenzymes of catalase. Among them, the samllest and most abundant species in cell-free extract, catalase-3 which also exhibited peroxidase activity was purified to electrophoretic homogeneity (145-fold purification) by chromatographic procedures. Its molecular weight was 155 kDa composed of four 38 kDa subunits. The $K_{m}$ value of catalase-3 for H$\_$2/O$\_$2/ was approximately 0.5 mM. This enzyme showed a typical ferric heme spectrum with maximum absorption at 405 nm. Upon binding to cyanide, the 405 nm peak shifted to 420 nm. Catalase-3 was very sensitive to inhibitors of heme proteins, such as cyanide, azide and hydroxylamine. A ratio of A$\_$405/A$\_$28O/ was 0.5 Catalase-3 was active over a wide range of pH, between pH 7 and 10. The enzyme was rather heat-labile and partially sensitive to edthanol-chloroform treatment, but resistant to 3-amino-1, 2, 4-triazole. Catalase-3 of D. radiophilus, which is a bifunction catalatic peroxidatic enzyme seemed to share certain molecular properties with the typical catalase and the catalase-[roxidase along with its own unique features.

• BMB Reports
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• 제31권2호
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• pp.144-148
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• 1998

A bifunctional catalase-peroxidase, designated catalase-2, of a UV resistant Deinococcus radiophilus was purified to electrophoretic homogeneity by both chromatographic and electrophoretic methods. Its molecular weight was 310 kDa and composed of a tetramer of 80 kDa subunits. The catalase-2 exerted its optimal activity at $30^{\circ}C$ and around pH 9. Its $K_m$ value for $H_{2}0_{2} $ was about 10 mM. It showed the typical ferric heme spectrum with maximum absorption at 403 nm which shifted to 419 nm in the presence of cyanide. The ratio of A40i' A2S0 was 0.48. Fifty percent inhibition of the enzyme activity was observed at $4.6{\times}10^{-6}$, $7.7{\times}10^{-6}$, and $3.0{\times}10^{-6}$ M of NaCN, $NaN_3$, and $NH_{2}OH$, respectively. The enzyme was thermostable and not sensitive to 3-amino-1,2,4-triazole. Treatment of the enzyme with ethanol-chloroform caused a partial loss (30%) of its activity. The catalase-2 was distinct from the Deinococcal bifunctional catalase-3 in a number of properties, particularly in its molecular structure and substrate affinity.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.193-198
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• 1988

Aspergillus niger KUF-04에서 얻은 catalase는 gel 여과를 포함하여 5단계를 걸쳐 정제하였으며, 9% 회수율로 64배 정제되었다. 본 효소는 406, 503, 625nm에서 흡광을 나타내었으며, 특히 406nm에서 뚜렷한 흡수대를 보여주었다. 이 효소 활성의 최적 pH와 온도는 각각 7.0과 6$0^{\circ}C$이었다. 이 효소는 pH4.0과 8.0 사이에서 안정했으며 열에 대한 안정성은 2$0^{\circ}C$에서 5$0^{\circ}C$까지는 안정했으나, 8$0^{\circ}C$에서 20분간 반응시켰을 때 효소의 활성이 전부 소실되었다. 이 효소의 활성은 주로 hydroxylamine, potassium cyanide, sodium azide에 의해 저해되었다.

• Journal of Microbiology
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• 제44권2호
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• pp.185-191
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• 2006

The photosynthetic bacterium, Rhodospirillum rubrum S1, when grown under anaerobic conditions, generated three different types of catalases. In this study, we purified and characterized the highest molecular weight catalase from the three catalases. The total specific catalase activity of the crude cell extracts was 88 U/mg. After the completion of the final purification step, the specific activity of the purified catalase was 1,256 U/mg. The purified catalase evidenced an estimated molecular mass of 318 kDa, consisting of four identical subunits, each of 79 kDa. The purified enzyme exhibited an apparent Km value of 30.4 mM and a Vmax of 2,564 U against hydrogen peroxide. The enzyme also exhibited a broad optimal pH $(5.0{\sim}9.0)$, and remained stable over a broad temperature range $(20^{\circ}C{\sim}60^{\circ}C)$. It maintained 90% activity against organic solvents (ethanol/chloroform) known hydroperoxidase inhibitors, and exhibited no detectable peroxidase activity. The catalase activity of the purified enzyme was reduced to 19 % of full activity as the result of the administration of 10 mM 3-amino-1,2,4-triazole, a heme-containing catalase inhibitor. Sodium cyanide, sodium azide, and hydroxylamine, all of which are known heme protein inhibitors, inhibited catalase activity by 50 % at concentrations of $11.5{\mu}M,\;0.52{\mu}M,\;and\;0.11{\mu}M$, respectively. In accordance with these findings, the enzyme was identified as a type of monofunctional catalase.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1994년도 한국생물과학협회 학술발표대회
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• pp.221.2-221
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• 1994

No Abstract, See Full Text

• Preventive Nutrition and Food Science
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• 제7권1호
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• pp.28-32
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• 2002

A Micrococcus sp. producing catalase was isolated from soil, and a commercial-scathe cultivation and purification of catalase were conducted. The maximum catalase activity was about 103 BU/mL obtained after 46 hr of cultivation in a 30 L fermenter containing 2% glucose, 2% peptone, 4% yeast extract, and 0.5% NaCl. Soybean sauce, CSL (corn steep liquor), and yeast extract were also studied as media substitutes in the media 30 L fermenter. The optimum medium components for the production catalase were found to be 2% glucose, 4% soybean sauce, and 16% CSL. In a 18 kL fermenter, the stationary phase in the cell growth and maximum catalase activity (112 BU/mL) were reached after 46 hr of cultivation, which was the same result as in the 30 L fermenter. The catalase activity was purified with over 17 folds in four steps with a 33.6% yield. From 104,250 mg of protein after cell lysis, 1,966 mg of the purified enzyme with a specific activity of 192.7 kBU/mg was obtained. The residual activity with the addition of 10% NaCl exhibited more than 100%. The use of just NaCl produced a higher residual activity than combination of bencol (benzyldimethyl ammoniumchloride) and PG (propyleneglycol).

• 한국동물학회:학술대회논문집
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• 한국동물학회 1999년도 한국생물과학협회 학술발표대회
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• pp.241.2-241
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• 1999

No Abstract, See Full Text

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.221.1-221.1
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• 2002

• Journal of Microbiology and Biotechnology
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• 제17권9호
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• pp.1460-1468
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• 2007

In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

• 한국미생물·생명공학회지
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• 제26권6호
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• pp.523-528
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• 1998

본 연구에서는 토마토에서 bacteriocin 생산균주를 분리하여 균주동정 및 bacteriocin 정제를 행하였다. 분리균주는 Gram 염색, catalase test, 포자생성유무, API 20 STREP. kit 등을 통하여 Enterococus faecium으로 잠정 동정하였다. 분리균주가 생산하는 항균성물질은 Gram 양성 세균인 Listeria manocytogmes, Leuonostoc mesenteroides, Lactobacillus plantarum, Streptococus agalartiae, Streptococus pyrogenes, Gram 음성 세균인 Pseudomonas aeruginosa등에 대해서 항균력을 나타내었다. 단백질 분해 효소인 proteinase K에 의해서 그 활성이 소실되고, catalase에 의해서는 항균력이 유지되므로 단백질성 bacteriocin임을 확인할 수 있었다. Bacteriocin 정제를 위하여 ethanol침전, CM Sepharose CL-6B 양이온교환 chromatography, Sephacryl S-100 HR gel filtration의 순으로 정제하여 초기 배양액의 bacteriocin활성 대비 35.8배의 정제도를 얻었다. 13% SDS-polyacrylamide gel전기 영동을 통하여 단일 band를 확인할 수 있었으며 분자량은 약 51 kDa 정도_로 추정되었다. Crude bacteriocin의 열 안정성은 10$0^{\circ}C$에서 1시간 처리하였을 때의 잔존활성이 3.3%로 비교적 낮았으며, pH에 대해서는 pH2~7에서 비교적 안정하였다. Bacteriocin의 항균작용을 알아보기 위하여 피검균인 Leu. mesenteroides를 초기농도 1.0$\times$$10^{9}$ CFU/$m\ell$로 접종한 배지에 bacteriocin을 첨가하여 배양하였을 때 4시간 이후에 완전히 사멸되므로 bacteriocidal 작용을 하는 것으로 추정되었다.