지방분해효소 생산균 Pseudomonas sp. OME 의 분리 동정 및 배양조건 최적화 Isolation and Identification of Lipolytic Enzyme Producing Pseudomonas sp. OME and Optimization of Cultural Conditions원문보기
폐식용유에서 지방분해효소를 생산하는 세균을 분리하였고, PIBWIN 세균동정 방법으로 생리 생화학적 특성을 조사하여 확인한 결과 Pseudomonas sp. OME로 동정하였다. 여러 기질로 지방분해효소 생산을 조사한 결과 올리브유에서 6.1 U/ml의 생산력을 나타내었다. 물리적 인자인 배양시간, 온도. pH 및 올리브유와 효모 추출액의 영양인자에 의한 지방분해효소 생산 조건을 조사 하였다. 효소의 분비는 배양시간. 올리브유 와 효모 추출액의 농도에 강한 영향을 받았으며, RSM을 이용한 최적화는 이들 인자를 가지고 조사하였다. RSM을 이용한 지방분해효소 생산은 배양시간. 올리브유와 효모 추출액의 농도가 48 hr, 0.3 g, 및 0.9 ml에서 최적 생산조건을 나타냈다.
폐식용유에서 지방분해효소를 생산하는 세균을 분리하였고, PIBWIN 세균동정 방법으로 생리 생화학적 특성을 조사하여 확인한 결과 Pseudomonas sp. OME로 동정하였다. 여러 기질로 지방분해효소 생산을 조사한 결과 올리브유에서 6.1 U/ml의 생산력을 나타내었다. 물리적 인자인 배양시간, 온도. pH 및 올리브유와 효모 추출액의 영양인자에 의한 지방분해효소 생산 조건을 조사 하였다. 효소의 분비는 배양시간. 올리브유 와 효모 추출액의 농도에 강한 영향을 받았으며, RSM을 이용한 최적화는 이들 인자를 가지고 조사하였다. RSM을 이용한 지방분해효소 생산은 배양시간. 올리브유와 효모 추출액의 농도가 48 hr, 0.3 g, 및 0.9 ml에서 최적 생산조건을 나타냈다.
Lipolytic enzyme-producing bacteria were isolated from edible oil mill effluents on tributyrin agar medium. The shake-flask-scale studies yielded a promising isolate and it was identified as Pseudomonas sp. An OME using various microbiological observations such as cultural, microscopic, and biochemi...
Lipolytic enzyme-producing bacteria were isolated from edible oil mill effluents on tributyrin agar medium. The shake-flask-scale studies yielded a promising isolate and it was identified as Pseudomonas sp. An OME using various microbiological observations such as cultural, microscopic, and biochemical tests was undertaken and confirmed using PIBWIN bacterial identification software. Lipolytic enzyme production was screened with oils such as sunflower, caster, coconut, tributyrin, and olive. Amongst these, olive oil showed an increased lipase production 6.1 U/ml. In view of the highest lipolytic enzyme production with olive oil, further optimizations were carried out using olive oil as a carbon source. Lipolytic enzyme production was optimized by a conventional 'one variable at a time' approach and the significant factors were further analyzed statistically using response surface methodology (RSM). The effect of physical factors such as incubation time, temperature, initial medium pH, and nutritional factors such as concentration of olive oil and yeast extract were examined for lipase production. Lipolytic enzyme secretion was strongly affected by three variables (incubation time, concentration of yeast extract and olive oil). Therefore, the interaction of these three factors was further optimized using response surface methodology. The optimized conditions of lipase production using response surface methodology yielded a maximum of 9.62 U/ml with optimum conditions for incubation, yeast extract and olive oil concentrations were found to be 48 hr, 0.3 g. and 0.9 ml. respectively.
Lipolytic enzyme-producing bacteria were isolated from edible oil mill effluents on tributyrin agar medium. The shake-flask-scale studies yielded a promising isolate and it was identified as Pseudomonas sp. An OME using various microbiological observations such as cultural, microscopic, and biochemical tests was undertaken and confirmed using PIBWIN bacterial identification software. Lipolytic enzyme production was screened with oils such as sunflower, caster, coconut, tributyrin, and olive. Amongst these, olive oil showed an increased lipase production 6.1 U/ml. In view of the highest lipolytic enzyme production with olive oil, further optimizations were carried out using olive oil as a carbon source. Lipolytic enzyme production was optimized by a conventional 'one variable at a time' approach and the significant factors were further analyzed statistically using response surface methodology (RSM). The effect of physical factors such as incubation time, temperature, initial medium pH, and nutritional factors such as concentration of olive oil and yeast extract were examined for lipase production. Lipolytic enzyme secretion was strongly affected by three variables (incubation time, concentration of yeast extract and olive oil). Therefore, the interaction of these three factors was further optimized using response surface methodology. The optimized conditions of lipase production using response surface methodology yielded a maximum of 9.62 U/ml with optimum conditions for incubation, yeast extract and olive oil concentrations were found to be 48 hr, 0.3 g. and 0.9 ml. respectively.
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제안 방법
Three factors selected from ‘one variable at a time’ approach, were further optimized and used in designing the experiment: Incubation time (A), Olive oil (B) and Yeast extract concentration (C).
이론/모형
The lipas activity in the fermented broth medium was measured by centrifugation of fermented culture at 10,000 rpm for 15 min at 4℃ and the crude centrifugal supernatant was used as enzyme source. Determination of lipase activity was done by the titrimetric method [14,16]. The emulsion of olive oil (10%, v/v) was prepared in arabic gum (5%) in sodium phosphate buffer (0.
1. Screening of lipase production by selected bacterial isolates using tributyrin agar plate assay method.
성능/효과
The values of the analysis of variance (ANOVA) for the fitted model are given in Table 6. The coefficient of determination (R2) for lipase production, as a function of independent variables, was 81%, which indicated that the model correlated well with the measured data and was statistically significant. The probability of the model equation implying that the model is significant.
3% (Table 4). The data suggested that the incubation period and the concentrations of olive oil and yeast extract was significantly influenced enzyme production when incubation temperature and medium pH kept constant.
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