• 제목/요약/키워드: Bacteriolytic enzyme

검색결과 9건 처리시간 0.016초

고정화 시스템을 이용한 용균효소의 생산

  • 류병호;박종옥;진성현
    • 한국미생물·생명공학회지
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    • 제24권4호
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    • pp.500-506
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    • 1996
  • Bacillus subtilis SH-1 screened from coastal sea water of South Korea was used to produce bacteriolytic enzyme. The production of bacteriolytic enzyme by immobilized cells was investigated. The optimum conditions for the continuous production of the bacteriolytic enzyme using immobilized cells were 2.4 mm diameter of 0.3% alginate beads, 20 ml/h of substrate feeding rate and 20 l/min of aeration rate. A productivity of 76.5 to 88.0 units/ml could be obtained for 25 days by continuous column reactor under the optimum conditions.

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호알칼리성 Bacillus sp.가 생산되는 Bacteriolytic Enzyme을 이용한 Bacillus subtilis의 형질전환 (Genetic Transformation of Bacillus subtilis by the Bacteriolytic Enzyme from Alkafophilic Bacillus sp.)

  • 유주현;이인숙;옥승호;박희경;염도영;배동훈
    • 한국미생물·생명공학회지
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    • 제21권5호
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    • pp.453-460
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    • 1993
  • The extracellular bacteriolytic enzyme from alkalophilic Bacillus sp. YJ-451 was endopeptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan. Protoplast transfomation system of B. subtilis by the lytic enzyme that differs, in mechanisms, from lysozyme which was used to transformation of B. subtilis was investigated. High protoplast yield was obtained from cells cultured in PAB at the late logarithmic growth phase.

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해양에서 용균효소를 분비하는 균주의 분리와 동정 (Isolation and Identification of a Bacteriolytic Enzyme-producing Bacterial Strain from Pusan Coastal Sea)

  • 진성현;류병호
    • 한국미생물·생명공학회지
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    • 제23권5호
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    • pp.580-587
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    • 1995
  • In order to produce the bacteriolytic enzyme, bacterial strains capable of excreting a large amount of the enzyme were screened from the coastal sea water samples in Pusan. The bacterial strain SH-1, which showed the highest activity among 43 bacteriolytic enzyme producing bacteria, was finally selected for further studies. The strain SH-1 was an endospore-forming grampositive rod, and the position of spore was paracentral. These morphological characteristics assigned the isolated strain to the morphological group I classified by Gordon. The fatty acid composition of the bacterial stain was analyzed to be consisted of branched chains of iso-Cn and anteiso-Cn. Based on the percent content of the branched chain (93.85%), the isolates could be identified as a species of Bacillus. According to the experimental results of the API system (API 50CHB & API 20E) the strain was identified as Bacillus subtilis. Numerical texonomy, in which 82 major characters were examined using several species of Bacillus as the standard bacteria, indicated that the strain SH-1 showed 90% similarity to Bacillus subtilis. Thus, the isolated strain SH-1 could be identified as Bacillus subtilis.

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Cloning and Expression in Escherichia coli of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Jung, Myeong-Ho;Park, Hee-Kyoung
    • Journal of Microbiology and Biotechnology
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    • 제2권3호
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    • pp.161-165
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    • 1992
  • The gene encoding the bacteriolytic enzyme cell wall peptidoglycan hydrolase from alkalophilic Bacillus sp. was cloned in E. coli using pBR322 as a vector. A recombinant plasmid, designated pYTR451, was isolated and the size of the cloned HindIII fragment was found to be 4.8 Kb. The cell wall hydrolysis activity of an extract of the E. coli harboring the recombinant plasmid pYTR 451 was detected by SDS- polyacrylamide gel containing 0.2% (w/v) purified cell wall of Bacillus sp. The molecular weight of the enzyme was estimated to be about 27, 000 corresponding to the molecular weight of the Bacillus sp. bacteriolytic enzyme. The recombinant plasmid was found to contain the fragment originated from Bacillus sp. YJ-451 chromosomal DNA by Southern hybridization.

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Nucleotide Sequence of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.

  • Jung, Myeong-Ho;Ohk, Seung-Ho;Yum, Do-Young;Kong, In-Soo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.73-77
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    • 1993
  • The nucleotide sequence of Bacillus sp. bacteriolytic enzyme gene, lytP and its flanking regions were determined. A unique open reading frame for a protein of Mw. 27, 000, and a putative terminator sequence, were found behind a concensus ribosome binding site located 8 nt upstream from ATG start codon. The primary amino acid sequence deduced from nucleotide sequence revealed a putative protein of 255 amino acid residues with an Mw. of 27, 420. No significant homology could be found between the amino acid sequence of Bacillus sp. bacteriolytic enzyme and that of other cell wall hydrolases.

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해양에서 분리한 Bacillus subtilis SH-1이 분비하는 용균효소의 정제 및 특성

  • 진성현;정영기;류병호
    • 한국미생물·생명공학회지
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    • 제24권2호
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    • pp.191-196
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    • 1996
  • The bacteriolytic enzyme produced from Bacillus subtilis SH-1 was purified and characterized, and its molecular weight was determined. The bacteriolytic enzyme activity was increased about 66.5 times via purification with recovery yield of 18.5%. The optimum pH and temperature of this enzyme were 9.0 and 50$\circ$C. The enzyme was stable within a pH range of 6.0-10.0 and unstable above 60 . The molecular weight of the enzyme was estimated to be 23,000 dalton in a form of monomer with no other subunits. Effect of the enzyme on the lysis of bacteria engaged in food posion was tested. The lysis degree was below 31% against Gram negative bacteria and above 48% in Gram positive bacteria. The values higher than 73% were obtained against Vibrio sp. and Listeria sp. As the turbidity of dissolved peptidoglycan clecreases, the free amino group levels were increased. And, based on hydrolysis of casein, this enzyme was thought to be an endopeptidase.

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Purification and Characterization of a Bacteriolytic Enzyme from Alkalophilic Bacillus sp.

  • Jung, Myeong-Ho;Kang, In-Soo;Bai, Dong-Hoon;Yu, Ju-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제1권2호
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    • pp.102-110
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    • 1991
  • Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

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해양에서 분리한 용균세균인 Bacillus subtilis SH-1의 배양특성 (Cultural Characterization of Bacteriolytic Bacillus subtilis SH-1 Isolated from Pusan Coastal Sea)

  • 류병호;진성현
    • 한국식품위생안전성학회지
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    • 제10권4호
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    • pp.231-237
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    • 1995
  • 해양에서 분리한 용균활성이 우수한 균주인 Bacillus subtilis SH-1의 배양특성을 조사하여 다음과 같은 결과를 얻었다. Bacillus subtilis SH-1에 의한 용균효소 생산을 위한 치적조건인 1.0% glucose, 1.0% yeast extract, 1.0% NaCl, 0.02% $K_2HPO_4,\;0.002%\;MgSo_4{\cdot}7H_2O,\;0.001%\;MnSO_4{\cdot}5H_2O,\;0.0001%\;FeSO_4{\cdot}7H_2O\;(ph\;8.0)$의 배지로 $30^{\circ}C$에서 진탕배양하여 배양 28시간일 때 효소활성이 가장 높았다.

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Staphylococcus aureus Phage Type 29에 대한 오골계 난백 Lysozyme의 용균성 (Lytic Action of Egg White Lysozyme Isolated from Ogol Fowl on Staphylococcus aureus Phage Type 29)

  • 오홍록;이종수;김찬조
    • 농업과학연구
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    • 제14권2호
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    • pp.286-294
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    • 1987
  • 오골계 난백 lysozyme의 용균특성을 조사하고, 이를 식품보존료로 이용하기 위한 자료를 얻고자 오골계 난백으로부터 추출 분리한 lysozyme의 각종 미생물에 대한 용균성을 조사하였다. 그 결과, 공시균중 Gram 양성균인 Staphylococcus aureus phage type 29와 Bacillus subtilis ATCC 6633에 대하여는 용균성이 인정되었으나, Gram 음성균인 E. coli를 비롯한 여타의 균종과 Staphylococcus aureus phage type 57은 lysozyme에 대한 용균 감수성이 인정되지 않았다. 분리된 lysozyme은 S. aureus phage type 29를 공시균주로 하여 이를 육즙배지에 접종하고, $37^{\circ}C$에서 24시간 정치배양하여 세포를 회수한 후 540nm에서 흡광도가 0.6이 되도록 0.05M 초산완충액(pH 4.5)에 현탁시켜, 여기에 0.05%의 lysozyme을 가하여 $30^{\circ}C$, 30분간의 조건으로 반응시킬 때 용균성이 가장 좋았다. 또한 0.05%의 lysozyme은 반응액 중에 glycine(1%)을 첨가하므로서 공시균에 대한 용균효과가 양자의 상승작용에 의하여 그 단용시보다 현저히 증대(<50%) 되었으나, 기타 다른 첨가제와 금속이온 및 lysozyme과의 혼용효과는 인정되지 않았다.

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