• Title/Summary/Keyword: cellulose filter

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Purification and Characterization of a Thermophilic Cellulase from a Novel Cellulolytic Strain, Paenibacillus barcinonensis

  • Asha, Balachandrababu Malini;Revathi, Masilamani;Yadav, Amit;Sakthivel, Natarajan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1501-1509
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    • 2012
  • A novel bacterial strain, MG7, with high cellulase activity was isolated and identified by morphological characteristics and molecular phylogeny analysis as Paenibacillus barcinonensis. Maximum production of cellulase by MG7 was observed at pH 7.0 and $35^{\circ}C$. The enzyme was purified with a specific activity of 16.88 U/mg, the cellulase activity was observed in a zymogram, and its molecular mass (58.6 kDa) was confirmed by SDS-PAGE. The purified enzyme showed maximum activity at pH 6.0 and $65^{\circ}C$ and degraded cellulosic substrates such as carboxy methyl cellulose (CMC), Avicel, filter paper, and ${\beta}$-glucan. The enzyme showed stability with 0.5% concentration of various surfactants. The $K_m$ and $V_{max}$ of cellulase for CMC and Avicel were found to be 0.459mg/ml and 10.46mg/ml/h, and 1.01 mg/ml and 10.0 mg/ml/h, respectively. The high catalytic activity and its stability to temperature, pH, surfactants, and metal ions indicated that the cellulase enzyme by MG7 is a good candidate for biotechnological applications.

Protein kinase C와 이와 관련된 단백질 연구

  • 이재란;김진한;최명언
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.135-135
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    • 1993
  • 신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2+}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

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Hydrolysis of Empty Fruit Bunch of Oil Palm Using Cellulolytic Enzymes from Aspergillus terreus IMI 28243

  • Kader, Jalil;Krishnasamy, Getha;Mohtar, Wan;Omar, Othman
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.514-517
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    • 1999
  • Hydrolysis of EFB (empty fruit bunch) derived from oil palm was studied using crude enzyme from Aspergillus terreus IMI 282743 along with commercial enzymes from Trichoderma reesei and Aspergillus niger. Hydrolysis at $40^{\circ}C$ and $50^{\circ}C$ with $\alpha$-cellulose or EFB gave significantly lower yield when commercial enzymes of T. reesei and A. niger were used and the hydrolysis time extended beyond 10 h. After 24 h of hydrolysis at $40^{\circ}C$ and $50^{\circ}C$, the filter paper activity (Fpase) from A. terreus retained as much activity as A. niger and it was significantly higher than T. reesei. Glucose concentration of 0.25% and 0.5% caused significant inhibition in the crude enzyme, but in regards to the commercial enzymes it only showed a slight effect. Crude enzymes from A. terreus could produce the highest reducing sugars when compared to commercial enzymes from T. reesei or A. niger. Nevertheless, low yield of sugar was observed for EFB for all treatments.

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Substrate Specificity of Cellulase from Aspergillus niger (Aspergillus niger가 생산하는 섬유소 분해효소의 기질에 대한 특이성)

  • Oh, Tae-Kwang;Park, Kwan-Hwa;Shin, Hyun-Kyung;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.162-166
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    • 1985
  • Three isozymes of Carboxymethyl Cellulase $(FI^*,\;FII^*,\;FIII)$ and two fractious of ${\beta}-1,4-D-Cellobiohydrolase$(CI, CIl) from Aspergillus niger were purified by Sephadex G-150, DEAE-Sephadex and Sephadex G-75 column chromatography. From the results of enzymatic hydrolysis and X-ray diffraction, ${\beta}-1,4-D-Cellobiohyarolase$ has a high activity toward highly crystalline cellulose such as filter paper and acts synergistically with Cx enzyme.

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Comparative study of clearing methods for air-borne asbestos analysis (공기 중 석면표준시료를 이용한 전처리 방법 비교 연구)

  • Yi, Gwang Yong;Shin, Jeong Ah;Jeong, Jee Yeon;Lee, In Seop;Park, Doo Yong
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.21 no.2
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    • pp.99-102
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    • 2011
  • Objectives: Acetone/triacetine method for clearing cellulose ester membrane (CEM) filter has been a popular method for air-borne asbestos analysis. However, as a weakness of this method, it is time consuming to analyses asbestos samples after sampling. Crystalclear method can be used to analyses asbestos samples promptly after sampling. Although a strength of crystal clear method exists, there was little valid studied for the method. This study was conducted to compare acetone/triacetine method with crystalclear one for analysing asbestos sample. Methods: Test samples made in three different concentration ranges(low, medium and high concentration) were analysed by phase contrast microscopy after acetone/triacetine and crystalclear method treatment respectively. Results: We did not find statistical difference in analysed results between two methods, which were conducted in three different concentrations ranges. Conclusions: We concluded that crystalclear method can be used as clearing method for air-borne asbestos analysis instead of acetone/triacetine method.

Development of an Integrated Oil Purification System (통합형 오일 정제 시스템의 개발)

  • Hong, Sung-Ho;Lee, Kyung-Hee;Jeong, Nam-Wha
    • Tribology and Lubricants
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    • v.38 no.4
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    • pp.121-127
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    • 2022
  • This study presents the development of an integrated oil purification system consisting of moisture removal, oil flushing, and oil filtering devices. In this system, the oil flushing device is combined with a micro-bubble generator. Oil purification is necessary for ensuring the high performance of the lubricant through the efficient removal of contaminants and thus enables good maintenance of mechanical systems. The developed purification system removes moisture, varnish, and solid particles. Moreover, during oil purification, the oil flushing device separates foreign materials and contaminants remaining in the lubricating oil piping or mechanical systems. The microbubble generator, which is combined with the oil flushing device, can separate harmful contaminants, such as sludge, wear particles, and rust, from piping or lubrication systems through the cavitation effect. Moisture is removed using a double high-vacuum chamber, while sludge and varnish are removed via electro-absorption using a high-voltage generator. Additionally, the total maintenance cost of the system is reduced through the use of domestically fabricated cartridge filters composed of glass fiber and cellulose. The heater, which maintains the temperature of the lubricant at 60℃, can process 41,000 L of lubricant simultaneously. Multiple tests confirmed that the proposed integrated purification system exhibits good performance in oil flushing and removal of water and varnish.

Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Effect of Sampling and Analytical Methods on the Fibrous Materials from the Ground Water (시료 채취 조건 및 검사방법에 따른 지하수내 섬유상 물질 검출 양상에 관한 연구)

  • Kim, Ji-Yong;Kim, Jung Ran;Cheong, Hae-Kwan;Lim, Hyun-Sul;Paik, Nam-Won
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.7 no.2
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    • pp.209-222
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    • 1997
  • Authors surveyed the ground water near the waste disposed from a fiberglass production factory to confirm the presence of glassfiber in the water and to determine the effect of sampling conditions and storage on the recovery of fibrous materials in the ground water. Sample was collected at every 4 hours for 48 hours consecutively. After finishing the 48 hours sample, water sampling was done from each tap after repeated turning on and off the water for 30 seconds at each time. Sample was collected in the two 1.5 liter polyethylene bottle after vigorously shaking the bottle with the same water several times with the flowing tap water. At each paired sample, one bottle was stored stand still at room temperature, and the other sample was filtered immediately after sampling. Water was filtered on the Mixed Cellulose Ester filter with negative pressure. Each sample was divided into upper and lower layer. The other bottle was stored at room temperature standstill for 7 days and filtered in the same fashion as the other pair of sample did. Each MCE filter was divided into 4 pieces and one piece was treated with acetone to make it transparent. Each prepared sample was observed by two researchers under the light and polarizing microscopy, scanning electron microscopy and energy dispersive X-ra microanalysis. Fibers were classified by the morphology and polarizing pattern under the polarizing microscope, and count was done. 1. There was a significant fluctuation in number of the fibers, but there was no specific demonstrable pattern. 2. Non-polarizing fibers frequently disappeared after 7 days's storage. But cluster of fibers were found at the wall of the same container by scratching technique. 3. Polarizing fibers were usually found in between the filter and the manicure pasted area. Possible explanations for this phenomenon will be that either these fibers are very light or have electronic polarity. Hence, these fibers are not able to be attached on the surface of slide glass. 4. Under the scanning electron microscopic examination, the fibers which are not refractive under the light microscopy were identified as glassfiber. Other fibers which is refractive under the polarizing microscopy were identified as magnesium silicate fibers. It is strongly suggested that development of standardized method of sample collection and measurement of fibrous material in the water is needed.

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Improved Organic Removal Efficiency in Two-phase Anaerobic Reactor with Submerged Microfiltration System (침지형 정밀여과시스템을 결합한 이상 혐기성 시스템에 의한 유기물 제거율의 향상)

  • Jung, Jin-Young;Chung, Yun-Chul;Lee, Sang-Min
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.4
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    • pp.629-637
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    • 2000
  • A two-phase anaerobic reactor with a submerged microfiltration system was tested for its ability to produce methane energy from organic wastewater. A membrane separation system with periodic backwashing with compressed air was submerged in the acidogenic reactor. The cartridge type of microfiltration (MF) membrane with pore size of $0.5{\mu}m$ (mixed esters of cellulose) was tested. An AUBF (Anaerobic Upflow Sludge Bed Filter: 1/2 packed with plastic media) was used for the methanogenic reactor. Soluble starch was used as a substrate. The COD removal was investigated for various organic loading with synthetic wastewater of 5,000 mg starch/L. When the hydraulic retention time (HRT) of the acidogenic reactor was changed from 10 to 4.5 days, the organic loading rate (OLR) varied from 0.5 to $1.0kg\;COD/m^3-day$. When the HRT of the methanogenic reactor was changed from 2.8 to 0.5 days, the OLR varied from 0.8 to $5.8kg\;COD/m^3-day$. The acid conversion rate of the acidogenic reactor was over 80% in the 4~5 days of HRT. The overall COD removal efficiency of the methanogenic reactor showed over 95% (effluent COD was below 300 mg/L) under the highly fluctuating organic loading condition. A two-phase anaerobic reactor showed an excellent acid conversion rate from organic wastewater due to the higher biomass concentration than the conventional system. A methanogenic reactor combined with sludge bed and filter, showed an efficient COD and SS removal.

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In vitro Fermentation of Rumen Microorganisms Cultured in Medium Supplemented with Bacterio-mineral Water (BMW) Produced from Bio-reacted Swine Manure

  • Kim, Chang-Hyun;Park, Joong Kook;Lee, Gi Yeong;Seo, In Joon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1435-1439
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    • 2005
  • Bacterio-mineral water (BMW) produced from manure has been known to exert a number of positive effects on animal production and odor control. An experiment was conducted to examine the effects of BMW produced from bio-reacted swine manure on in vitro gas production, cellulose degradation, microbial growth and fibrolytic enzyme activities of mixed rumen microorganisms. The five levels of 0, 0.001, 0.005, 0.01 and 1.0% BMW were supplemented into serum vials containing mixed rumen microorganisms. Incubations were carried out anaerobically at $39^{\circ}C$ without shaking for 0, 12, 24, 48, 72 and 96 h. There were no significant (p>0.05) differences among the treatments for the initial rate of gas production. At 72 h incubation, the gas production tended (p<0.1) to be increased by the 0.01 and 1.0% BMW treatments compared with control and the 0.001% BMW treatment. At the end of incubation (96 h), the sample supplemented with 0.01% BMW was higher (p<0.05) than control (0% BMW) in the gas production. The microbial growth rate was increased by all the BMW treatments, while 0.01% BMW was most effective in stimulating the growth rate. Although the addition of BMW on the filter paper DM degradation was not significantly influenced throughout the incubation period except the 48 h incubation, DM degradation tended to be increased by all BMW treatments compared with control. The addition of both 0.005 and 0.01% BMW highly increased (p<0.05) CMCase activity compared with control after 24 h and 48 h incubation, while at the 72 h incubation the 0.01% BMW addition only significantly increased (p<0.05). After 72 h incubation, the xylanase activity was significantly (p<0.05) increased with the addition of 1.0% BMW compared with the addition of 0.001 and 0.005% BMW, while at the other incubation times, the xylanase activity was not different among the treatments. In conclusion, the 0.01% BMW of supplementation level would be the suitable addition level to stimulate rumen fermentation increasing microbial growth and cellulose degradation.