[논문]Optimization Using 33 Full-Factorial Design for Crude Biosurfactant Activity from Bacillus pumilus IJ-1 in Submerged Fermentation

Kim, Byung Soo; Kim, Ji Yeon

doi:10.4014/mbl.1908.08001

Optimization Using 33 Full-Factorial Design for Crude Biosurfactant Activity from Bacillus pumilus IJ-1 in Submerged Fermentation 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.48 no.1, 2020년, pp.48 - 56

Kim, Byung Soo (Department of Statistics, Institute of Statistical Information, Inje University) , Kim, Ji Yeon (Department of Liberal Arts, Inje University)

Abstract ▼ AI-Helper

This study aimed to optimize the culture conditions to improve the crude biosurfactant activity of Bacillus pumilus IJ-1, using a 33 full-factorial design of response surface methodology (RSM). It was found that submerged fermentation of B. pumilus improved the activity of the crude biosurfactant. The factors selected for optimization were NaCl concentration, temperature, and tryptone concentration. Response surface analysis revealed that the fitted quadratic model was statistically significant and produced an adequate R2 value (0.9898) and a low probability value (<0.0001). The optimum level for each factor was found to be 0.567% (w/v) NaCl, 21.851℃ and 0.765% (w/v) tryptone, respectively. Crude biosurfactant activity was found to be most affected by tryptone concentration; then temperature, and finally NaCl concentration. Our results may potentially facilitate large-scale biosurfactant production from B. pumilus IJ-1.

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문제 정의

We previously stated the consequence of medium composition and culture conditions on crude biosurfactant activity from Bacillus pumilus IJ-1, using an established method [23]. According to a literature survey, this paper is another report to elucidate optimization of culture conditions for crude biosurfactant activity from B. pumilus, using RSM at a flask level.

제안 방법

NaCl, tryptone, and culture temperature were determined as major factors affecting significant crude biosurfactant activity. Based on previous studies, appropriate models were devised to analyze factor interactions. For its validity, the parameter range was selected in accordance with the previous report [23].
For its validity, the parameter range was selected in accordance with the previous report [23]. Experiments were based on 3³ full-factorial design: NaCl (0, 0.5, 1.0%, w/v), tryptone (0, 0.5, 1.0%, w/v), and temperature (15, 20, 25℃) by B. pumilus IJ-1 inoculated with 1% (w/v) seed culture at pH 9.0 in shaking incubator (200 rpm) for 72 h.
RSM was required to ascertain the interactive effects of three factors at three levels. The consequent data was then subjected to multiple regression analysis on the basis of the empirical model. After completing the experiments, the relationship between the independent factors and their responses was fitted to the following predictive second-order polynomial equation:
The optimal values of the experimental conditions resulted from solving the regression equation and analyzing the response-surface contour plots. The responses to each run were subjected to multiple regression analysis with the help of SAS 9.
Statistical approaches facilitate the formulation of biosurfactant production media and may be crucial to enhance productivity [34]. This study attempted to optimize culture conditions for crude biosurfactant activity by B. pumilus IJ-1. Using a combination of a 3³ full-factorial design and RSM, we determined the optimal operating conditions for large-scale biosurfactant production by B.
Each nutrient source required an uninoculated control flask to measure the initial surface tension. To enhance accuracy, each measurement resulted from three separate experiments, and the surface tension of the sample was expressed into the mean value in each measurement.
To validate these measurements, calibration used distilled water (ST = 71.5 dyne/cm ± 0.5) before obtaining sample measurements.

대상 데이터

The design was comprised of 29 experimental trials including 27 for factorial design, and 2 for replication of the central points (Table 2). The reduction ratio of surface tension (%) was considered response values (Y).

데이터처리

2 software (SAS Institute, USA) to obtain the coefficients of the polynomial equation. The model was statistically validated via the analysis of variance (ANOVA) provided in the same software. The equation was then expressed in the form of a threedimensional response surface and their respective contour plots, using the software.
The optimal values of the experimental conditions resulted from solving the regression equation and analyzing the response-surface contour plots. The responses to each run were subjected to multiple regression analysis with the help of SAS 9.2 software (SAS Institute, USA) to obtain the coefficients of the polynomial equation. The model was statistically validated via the analysis of variance (ANOVA) provided in the same software.
The t-values and the corresponding p-values were calculated from the Student’s t distribution, along with the parameter estimates (Table 4).

이론/모형

Reducing the surface tension of the bacterial growth medium, the effects of biosurfactants can be indirectly measured by determining surface tension [26, 27]. The surface tension was determined on the crude biosurfactants with a Surface Tension Analyser DST-60 digital surface tensiometer (Surface Electro Optics, Korea), using the Du Nouy ring method at 20℃ [28]. To validate these measurements, calibration used distilled water (ST = 71.

후속연구

pumilus IJ-1. Therefore, further research is required to optimize the biosurfactant yield in specialized bioreactors.

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