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. 2018 Oct 23:9:2377.
doi: 10.3389/fmicb.2018.02377. eCollection 2018.

Production of Cold-Active Lipase by Free and Immobilized Marine Bacillus cereus HSS: Application in Wastewater Treatment

Sahar W M Hassan  1 , Hala H Abd El Latif  2 , Safaa M Ali  2
Affiliations

Production of Cold-Active Lipase by Free and Immobilized Marine Bacillus cereus HSS: Application in Wastewater Treatment

Sahar W M Hassan et al. Front Microbiol. .

Abstract

Lipases are enzymes that have the potential to hydrolyze triacylglycerol to free fatty acids and glycerol and have various applications. The aim of the present study was to isolate and screen marine bacteria for lipase production, optimize the production, and treat wastewater. A total of 20 marine bacterial isolates were obtained from the Mediterranean Sea and were screened for lipase production. All isolates were found to have lipolytic ability. The differences between the isolates were studied using RAPD-PCR. The most promising lipase producer (isolate 3) that exhibited the highest lipolytic hydrolysis (20 mm) was identified as Bacillus cereus HSS using 16S rDNA analysis and had the accession number MF581790. Optimization of lipase production was carried out using the Plackett-Burman experimental design with cotton seed oil as the inducer under shaking conditions at 10°C. The most significant factors that affected lipase production were FeSO4, KCl, and oil concentrations. By using the optimized culture conditions, the lipase activity increased by 1.8-fold compared with basal conditions. Immobilization by adsorption of cells on sponge and recycling raised lipase activity by 2.8-fold compared with free cells. The repeated reuse of the immobilized B. cereus HSS maintained reasonable lipase activity. A trial for the economic treatment of oily wastewater was carried out. Removal efficiencies of biological oxygen demand, total suspended solids, and oil and grease were 87.63, 90, and 94.7%, respectively, which is promising for future applications.

Keywords: Plackett–Burman; immobilization; lipase; recycling; wastewater treatment.

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Figures

FIGURE 1
FIGURE 1
Hydrolytic zone of Tween 80 produced by isolate 3.
FIGURE 2
FIGURE 2
Agarose gel shows polymorphism between isolates using primer 16S F (A) and 16S1100R (B).
FIGURE 3
FIGURE 3
Phylogenetic tree showing the incidental evolutionary relationships between different biological Bacillus species. The dendrogram was generated using the MEGA 5 program.
FIGURE 4
FIGURE 4
Effect of temperature (A), agitation (B), and different inducer substrates (C) on lipase activity of B. cereus HSS.
FIGURE 5
FIGURE 5
Elucidation of fermentation conditions affecting the production of lipase.
FIGURE 6
FIGURE 6
Response surface plots of the interaction between process variables in lipase production by B. cereus HSS.
FIGURE 7
FIGURE 7
Verification experiment: Lipase production by B. cereus HSS grown on basal versus optimized medium.
FIGURE 8
FIGURE 8
Production of lipase by entrapped cells (A) and adsorbed cells on different support materials (B), electron micrograph of sponge (C1) and cells adsorbed on sponge (C2), and recycling of adsorbed B. cereus HSS on sponge (D).
FIGURE 9
FIGURE 9
GC-MS chromatogram of the oily wastewater (A) control (untreated) and (B) treated with lipase producer B. cereus HSS after 72 h.
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