• 제목/요약/키워드: cell lytic enzyme

검색결과 52건 처리시간 0.03초

Bacillus sp. LM-8이 생산하는 Lactobacillus plantarum 용균 효소의 정제 및 효소 특성 (Purification and Enzyme Property of a Cell-Wall Lytic Enzyme Produced by Bacillus sp. LM-8 against Lactobacillus plantarum.)

  • 마호우;신원철
    • 한국미생물·생명공학회지
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    • 제30권1호
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    • pp.33-38
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    • 2002
  • Lactobacillus plantarum 용균 효소를 생산하는 균주를 배양하여 생산된 효소를 정제한 결과 비활성도가 5.8 units/mg protein 이었고 정제도 8.3배, 수율은 30%이었다. 정제효소의 분자량은 gel filtration과 SDS-polyacrylamide gel eletrophoresis를 이용하여 측정한 결과 60,000 kDa 이었다. 용균 효소의 최적 반응 시간은 20분이었으며 최적 온도는 4$0^{\circ}C$, 최적 pH는 3.0이었다. 온도 안정성은 각 온도에서 30분간 처리하였을 때 $30^{\circ}C$까지는 안정하였으나 4$0^{\circ}C$에서는 80% 활성을 나타내었다. 효소의 pH 안정성은 실온에서 1시간 처리하였을 때 pH 4~7에서 안정성을 나타내었다.

Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소 II. 효소활성에 미치는 기질 효모의 배양조건 및 전처리 효과 (Yeast Cell Wall Lytic Enzyme Produced by Dicyma sp. YCH-37 II. Effect of Culture Conditions and Pretreatment of Yeast on the Enzyme Activity)

  • 정희철;함병권;유주현;배동훈
    • 한국식품과학회지
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    • 제29권5호
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    • pp.1021-1027
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    • 1997
  • Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소의 성질을 검토한 결과, 각종 환원제와 금속이온에 대체로 안정하였고, guanidine-HCl을 제외한 여러 화학수식제에 대해서도 안정하였다. 배양시간, 전처리 및 배양조건에 따른 영향을 검토한 결과, 정지기 및 사멸기에 있는 효모보다는 대수증식기의 효모, 그리고 생효모에 비해 열처리된 효모가 더 잘 용균되었다. Butanol, acetone 등의 유기용매로 처리된 효모가 그렇지 않은 효모보다 용균도가 좋았으며, 0.5 M ammonium sulfate가 함유된 Yeast extract-Malt extract 배지에서 생육한 효모, 그리고 진탕배양한 효모보다 정치배양한 효모가 용균효소에 의해 더 잘 용균되었다. SDS, Triton X-100, ${\beta}-mercaptoethanol$, potassium chloride, sodium sulfite 등의 화학수식제를 효소반응액에 첨가하였을 때 기질 효모는 더 잘 용균되었다.

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Antialgal Effect of a Novel Polysaccharolytic Sinorhizobium kostiense AFK-13 on Anabaena flos-aquae Causing Water Bloom

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1613-1621
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    • 2006
  • Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

Snail Lytic Enzyme에 의한 전분리용성 효모 및 Saccharomyces cerevisiae의 원형질체 형성 (Protoplast Formation of the Amylolytic Yeast and Saccharomyces cerevisiae by Snail Lytic Enzyme from Helix pomatia)

  • 구영조;박완수;신동화;유태종
    • 한국미생물·생명공학회지
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    • 제13권2호
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    • pp.137-144
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    • 1985
  • 전분자원의 효율적 이용을 위한 방법의 일환으로 분리동정된 H. anomala var. anomala FRI YO-32와 S. cerevisiae와의 세포융합가능성을 검토하기 위하여 원형질체형성을 위한 기본적인 제반조건에 대하여 실험하였다. 세포벽분해효소로서 달팽이 (Helix pomatia) 추출 효소를 사용하여 원형질체의 형성시수율에 영향을 미치는 중요한 인자로서, 함유황화합물에 의한 전처리유무 및 이러한 화합물의 처리농도 및 처리방법, 세포벽분해효소의 농도 및 처리시간, 공시효모의 성장시기 및 효모세포의 수와 삼투압안정제 (KCI)의 농도 등이 고려되었으며, 원형질체형성을 위한 이러한 인자들의 최적처리조건이 검토되었다.

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세포벽 분해효소 처리에 의한 연잎 추출물의 항산화 및 tyrosinase 저해 활성 (Anti-oxidant and Whitening Effects of Cell Lytic Enzyme-treated Lotus Leaf Extract)

  • 최선주;김소영;이성철;이진만;이인숙;정문영;양삼만;채희정
    • KSBB Journal
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    • 제24권6호
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    • pp.579-583
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    • 2009
  • The effects of cell lytic enzyme treatment on total phenolic content, antioxidant and antityrosinase activities of lotus leaf were investigated. The dried lotus leaves were hydroyzed by cell lytic enzymes such as Promozyme, Ceremix, Pectinex, Ultraflo, Celluclast, Pentopan, Tunicase, Viscozyme at their optimum pHs (pH 5-8) at $50^{\circ}C$ for 4 hrs. Depending on the enzymes used, total phenolic compounds content was measured as $1,079-1,476{\mu}g$/mL, and antioxidant activities and whitening activities were increased by 5~10% and 20%, respectively Among the tested hydrolytic enzymes, Promozyme (pullulanase) was selected as the most suitable enzyme for the extraction of total polyphenol from lotus leaf. The optimal dosage of Promozyme were found to be 1-2% (w/w). By Promozyme treatment, total phenolic compounds content of the lotus extract significantly increased compared to the extraction without enzyme treatment.

Trichoderma koningii의 conidiospore로부터의 원형질체 분리에 관하여 (Isolation of protoplast from conidiospore of Trichoderma koningii)

  • 박희문;홍순우;하영칠
    • 미생물학회지
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    • 제21권4호
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    • pp.213-220
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    • 1983
  • Conditions for isolation of protoplasts from conidiospores of Trichoderma koningii ATCC 26113 were tested. Maximum production of conidial protoplasts was obtained by preincubation of conidiospores on liquid minimal medium for 8 1/2 hrs. and by reaction with cell wall lytic enzyme for 3 hrs. Among effective cell wall lytic enzymes (Driselase, p-Glucuronidase, Novozyme and Zymolyase), Driselase was the most effective one on the production of conidial protoplasts. The production of conidial protoplasts was also enhanced by addition of 2-Deoxy-D-Glucose $(25{\mu}g/ml)$ into liquid minimal medium. Over 70% of the initial swollen conidia, preincubated in liquid minimal medium supplemented with 2-Deoxy-D-Glucose $(25{\mu}g/ml)$, were converted to protoplasts by incubation with 2% (w/v) commercial lytic enzyme Driselase at $28^{\circ}C$ for 3 hrs. The reversion frequency of the conidial protoplasts was about 30 times (25-50%) higher than that of mycelial protoplasts (0.6-1.3%).

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Enzyme Profiles of Alga-Lytic Bacterial Strain AK-13 Related with Elimination of Cyanobacterium Anabaena cylindrica

  • Kim, Jeong-Dong;Han, Myung-Soo
    • 환경생물
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    • 제22권1호
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    • pp.184-191
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    • 2004
  • To investigate bacteria with algalytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles associated with alga-lytic activity, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Among 178 isolates, only nine isolates exhibited lytic abilities against A cylindrica on the agar plates, and then the isolate AK-13 was selected as the strongest in lysing the cyanobacterium A. cytindrica. The strain AK-13 was characterized and identified as Sinorhizobium sp. based on fatty acid methyl ether profiles and 16S rDNA sequence. According to the results of the enzyme assays, in the strain An-13 of Sinorhizobium sp., alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase was produced, namely CMCase, laminarinase and protease were highly active. None of glycosidase was produced. Therefore, enzyme systems of Sinorhizobium sp. AK-13 were very complex to degrade cell walls of A. cylindrica. The peptidoglycans of A. cylindrica mat be hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by Sinorhizobium sp. AK-13.

열 처리와 효소 처리에 의한 탈지 참깨박 단백질의 추출율 향상 (Improvement of protein extraction efficiency from defatted sesame meal with thermal and enzymatic treatments)

  • 인만진
    • Journal of Applied Biological Chemistry
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    • 제63권4호
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    • pp.291-295
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    • 2020
  • 참기름 제조의 부산물인 참깨박의 활용도를 높이기 위하여 효소 처리에 의한 참깨박 불용성 단백질의 추출 조건을 조사하였다. 단백질 분해효소인 Alcalase, Flavourzyme, Neutrase, Protamex의 처리 결과를 대조구와 비교한 결과 Protamex가 고형분과 단백질 함량 증가에 효과적이었다. Protamex의 반응 조건(50 ℃, pH 6.0)에서 효소의 사용량은 탈지 참깨박의 1%, 효소반응시간은 3시간이 적당하였다. 효소 처리 효율을 향상시키기 위하여 참깨박을 열처리하면, 열처리 온도가 증가할수록 추출되는 단백질 함량은 증가하였으며 110이상에서는 미미하게 증가하였다. 세포벽 분해효소(Tunicase)와 단백질 분해효소의 병용처리가 단백질 가용화에 미치는 효과를 조사한 결과, 단백질 분해효소를 처리한 다음에 세포벽 분해효소를 처리하는 것이 가장 효과적이었다. 열처리(110 ℃, 10분) 후 Protamex와 Tunicase를 순차적인 처리로 단백질 함량이 열처리와 효소처리하지 않은 대조구의 약 3.6배(9.85→35.58 mg/mL), 열처리만 실시한 대조구의 약 2.2배(15.83→35.58 mg/mL) 증가하였다.

Purification and Characterization of A Cell Wall Hydrolyzing Enzyme Produced by An Alkalophilic Bacillus sp. BL-29

  • Hong, Soon-Duck;Kim, Tae-Ho;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
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    • 제5권4호
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    • pp.206-212
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    • 1995
  • A strain BL-29, which produces a extracellular lytic enzyme on E. coli was isolated from the soil. The strain was identified as belonging to the genus Bacillus sp. The lytic enzyme was purified to homogeneity by ion exchange chromatography and gel filtration. Specific activity of the purified enzyme was 28, 850 U/mg protein and yield of the enzyme was 5$%$. The purified enzyme showed a single band on SDS-PAGE and its molecular weight was estimated to be 31, 000 by SDS-polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum temperature and pH were $55^{\circ}C$ and pH 10.0, respectively. The enzyme was stable at $45^{\circ}C$ but enzyme activity was reduced by up to 50$%$ when the temperature was raised to $55^{\circ}C$ for 15 min. Stable range of pH was from 5.0 to 11.0. but Enzyme activity was inhibited by lead-acetate, mercuric chloride, ethylene glycol-bis-[$\beta$-aminoethyl ether]-N, N, $N^1, $N^1$-tetraacetic acid (EGTA), and ethylenediamine tetraacetic acid (EDTA), but not affected considerably by treatment with other chemical reagents.

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호알카리성 Bacillus sp. 4830이 생산하는 Streptococcus mutans 세포벽 분해효소의 분리와 특성 (Production and Characteristics of Lytic Enzyme against Streptococcus mutans Cell Wall from Alkalophilic Bacillus sp. 4830)

  • 김윤근;배동훈
    • 한국식품과학회지
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    • 제35권6호
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    • pp.1143-1149
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    • 2003
  • 본 연구는 Streptococcus mutans를 특이적으로 용균하는 효소를 탐색하고 효소생산 균주의 동정을 수행하였다. 토양으로부터 5,000여주의 알칼리 내성 판주를 선별하였으며 그중 용균활성이 있는 22주를 선별하였으며 가장 용균활성이 높은 균주 1주를 선별하여 Strain No. 4830이라 명명한 후 효소의 생산 조건 및 효소의 특성을 확인한 결과 pH 5에서부터 pH 11에서 안정하였으며 pH 7.0 이상에서 Streptococcus mutans에 대하여 용균활성이 나타났다. 또한 $30^{\circ}C$ 이상에서 효소의 활성을 확인할 수 있었으며 $50^{\circ}C$에서 최대효소활성을 확인하였다. 효소의 열안정성은 $40^{\circ}C$ 이하에서는 안정한 것으로 확인되었다. 균주의 특성을 검토한 결과 균체의 지방산 조성은 중성 조건에서 생장하는 Bacillus계열과는 달리 16-C계열의 지방산함량이 상당히 높은 것을 확인하였으며 Obligate alkalophilic의 특성으로 알려진 branched 15-C 지방산인 $iso-C_{15:0}$$anteiso-C_{15:0}$의 함량이 매우 높은 것을 확인할 수 있었다. 또한 생화학적 특성으로 Bacillus 계열로 추정이 가능하였으며 다양한 종류의 당류에 대하여 acid를 형성하지 않은 것으로 확인되어 Strain No. 4830은 Obligate alkalophilic의 특성을 나타내었다. 또한 15% NaCl의 농도에서도 생장하는 것으로 보여 염에 대하여 내성을 갖는 것으로 나타났다. 16S rRNA 분석결과 Bacillus alcalophilus 균주와 94%의 유사성을 나타내었으나 생화학적 및 이화학적인 특성 등에서 Bacillus alcalophilus와 다른 균주로 동정되었다.