[논문]Improved Biosurfactant Production by Bacillus subtilis SPB1 Mutant Obtained by Random Mutagenesis and Its Application in Enhanced Oil Recovery in a Sand System

Bouassida, Mouna; Ghazala, Imen; Ellouze-Chaabouni, Semia; Ghribi, Dhouha

doi:10.4014/jmb.1701.01033

Improved Biosurfactant Production by Bacillus subtilis SPB1 Mutant Obtained by Random Mutagenesis and Its Application in Enhanced Oil Recovery in a Sand System 원문보기

Journal of microbiology and biotechnology, v.28 no.1, 2018년, pp.95 - 104

Bouassida, Mouna (Unite Enzymes et Bioconversion, Ecole Nationale d'Ingenieurs de Sfax, Universite de Sfax) , Ghazala, Imen (Laboratoire d'amelioration des plantes et valorisation des agroressources, Ecole Nationale d'Ingenieurs de Sfax, Universite de Sfax) , Ellouze-Chaabouni, Semia (Unite Enzymes et Bioconversion, Ecole Nationale d'Ingenieurs de Sfax, Universite de Sfax) , Ghribi, Dhouha (Unite Enzymes et Bioconversion, Ecole Nationale d'Ingenieurs de Sfax, Universite de Sfax)

Abstract ▼ AI-Helper

Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. Biodegradability and low toxicity have led to the intensification of scientific studies on a wide range of industrial applications for biosurfactants in the field of environmental bioremediation as well as the petroleum industry and enhanced oil recovery. However, the major issues in biosurfactant production are high production cost and low yield. Improving the bioindustrial production processes relies on many strategies, such as the use of cheap raw materials, the optimization of medium-culture conditions, and selecting hyperproducing strains. The present work aims to obtain a mutant with higher biosurfactant production through applying mutagenesis on Bacillus subtilis SPB1 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on blood agar and subsequent formation of halos, the mutated strains were examined for emulsifying activity of their culture broth. A mutant designated B. subtilis M2 was selected as it produced biosurfactant at twice higher concentration than the parent strain. The potential of this biosurfactant for industrial uses was shown by studying its stability to environmental stresses such as pH and temperature and its applicability in the oil recovery process. It was practically stable at high temperature and at a wide range of pH, and it recovered above 90% of motor oil adsorbed to a sand sample.

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문제 정의

In this work, we noted a good correlation between the blood agar lysis and the emulsification activity (Table 1). The isolates, which were positive in blood agar lysis, also gave a high emulsification property.
subtilis strain SPB1 by using a combination of nitrous acid and ultraviolet rays to induce mutation. This investigation aimed to develop a cost-effective and simple approach in order to increase the biosurfactant productivity. This work also describes the biosurfactant properties of the selected mutant under extreme environmental conditions, its biological activity, and its potential for industrial applications.
This investigation aimed to develop a cost-effective and simple approach in order to increase the biosurfactant productivity. This work also describes the biosurfactant properties of the selected mutant under extreme environmental conditions, its biological activity, and its potential for industrial applications.

가설 설정

Isolation of mutants, overproducing biosurfactant, with the blood agar plate test has been previously reported [32, 33]. In this study, we tested all the survivors for their hemolytic activity. As described by Beggs [34], the fewer the positive mutant cells, the more negative the mutant strains are in the survival cells.

제안 방법

subtilis SPB1 biosurfactant production was performed as reported by Ghribi and Chaabouni [16], with slight modifications. At time-defined intervals, samples were collected and submitted to analysis in order to determine biomass concentration and biosurfactants production. All experiments were performed in duplicates.
The purpose of the present work was to provide an economical methodology to enhance the biosurfactant production of B. subtilis strain SPB1 by using a combination of nitrous acid and ultraviolet rays to induce mutation. This investigation aimed to develop a cost-effective and simple approach in order to increase the biosurfactant productivity.

대상 데이터

Treatment of vegetative cells with UV rays.Mutagenesis with short wavelength ultraviolet irradiation was carried out using a Philips tube (15 W). The cell suspension (OD_600nm of 0.
The sand sample used in this study was collected from Tunisian Djerba beach. It was sifted to remove impurities and large particles and incubated overnight at 105°C to remove humidity [20].

성능/효과

In a conclusion, through random mutagenesis, by UV irradiation and nitrous acid treatments, a mutant with the ability to produce biosurfactant in a large amount compared with the wt was selected. The newly produced biosurfactant was effective over a wide range of temperatures and pHs, able to remove hydrophobic contaminants, and presented lower toxicity, thus justifying the possibility of its industrial use, especially in environmental application.
In conclusion, after three rounds of mutagenesis by nitrous acid applied respectively on the B. subtilis wild-type (wt) strain, the mutant M35, and finally the mutant M35-5, a mutant named B. subtilis M2 showed the highest hemolytic and emulsification improvement (109% and 118%, respectively). Therefore, it was selected for all further experiments, with more than 2-fold biosurfactant production.

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표제어: PCR

동의어: Packet Collision Rate

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