Enhancement of Halophilic Lipase Production by  Virgibacillus alimentarius LBU20907 using a Statistical Approach and  Scale-Up in a Fermenter

Sawitree DUERAMAE; Preeyanuch B OVORNREUNGROJ; Toshiki ENOMOTO; Duangporn KANTACHOTE

Authors

Sawitree DUERAMAE Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110
Preeyanuch B OVORNREUNGROJ Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110
Toshiki ENOMOTO Department of Food Science, Ishikawa Prefectural University, Nonoichi, Ishikawa
Duangporn KANTACHOTE Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110

Keywords:

Central composite design, halophilic lipase, Plackett-Burman design, response surface, methodology, Virgibacillus alimentarius

Abstract

Virgibacillus alimentarius LBU20907 is an efficient extracellular halophilic lipase producer. Isolate LBU20907 was identified by 16S rRNA gene sequence analysis and phenotypic identification before determining the optimal nutritional and cultural conditions for its halophilic lipase activity. Two statistical designs were used to predict the responses to varying factors: Plackett-Burman experimental design (PBD) and central composite design (CCD). The important factors affecting the halophilic lipase production were identified by 2-level PBD of 11 physicochemical parameters screened, 3 factors produced significantly positive effects. These were olive oil, CaCl₂, and temperature. The response surface methodology (RSM) was applied to the CCD to predict the most productive levels of these 3 factors. The results predicted that a maximum halophilic lipase activity of 65.96 U mL^-1 should be achieved with an olive oil concentration of 1.68 %, a CaCl₂ concentration of 0.14 g L^-1 and a temperature of 39.5 °C. The predicted halophilic lipase activity in the optimized medium represented an increase of nearly 42 % compared with the predicted result of 46.50 U mL^-1 in the non-optimized medium. The model was validated by subsequent experimentation at the optimized conditions. The predicted values were in agreement with the experimental values with a coefficient of determination (R²) of 0.9995. After scale-up using a 3 L laboratory fermenter, a halophilic lipase yield of 82.50 U mL^-1 was effectively achieved: 77 % more than predicted to occur in the non-optimized medium. Therefore, halophilic lipase production by V. alimentarius LBU20907 can be regarded as promising attractive catalysis for several industrial applications.

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