Molecular Detection and Optimization of Extracellular Alkaline Protease Production of Bacillus pseudofirmus MSB22 Using Response Surface Methodology

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Borhani M.S.; ETEMADIFAR Z.; EMTIAZI G.; JORJANI E.

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Journal: JOURNAL OF BIOTECHNOLOGY Year:2018 | Volume:9 | Issue:4 Page(s): 507-515

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Title

Molecular Detection and Optimization of Extracellular Alkaline Protease Production of Bacillus pseudofirmus MSB22 Using Response Surface Methodology

Author(s)

Borhani M.S. | ETEMADIFAR Z. | EMTIAZI G. | JORJANI E. | Issue Writer Certificate

Keywords

Alkaline ProteaseQ2

Bacillus pseudofirmusQ1

Polymerase Chain ReactionQ2

One-factor-at-atime methodQ1

Rotatable central composite designQ1

OptimizationQ2

Abstract

Aims Alkaline Protease is one of the most important groups of industrial enzymes with many applications. The aim of this study was to determine the physicochemical parameters affecting the production of Alkaline Protease enzyme produced by Bacillus pseudofirmus MSB22 by onefactor-at-a-time (OFAT) method and optimize the production of this enzyme by the response surface methodology (RSM) in the form of a Rotatable central composite design. Materials & Methods In the present experimental study, the isolation of microorganism producing Alkaline Protease from wastewater from sausage and lunch meat factories in Isfahan was carried out. The morphological and biochemical characteristics of the strain were performed according to the Bergey’ s book and amplification of 16S rRNA gene sequences. Detection of metalloproteinase gene and alkaline serine protease was done by Polymerase Chain Reaction (PCR) reaction and enzyme activity measurement was performed by Folin reagent. Screening of variables effective in enzyme production was done, using the one-factorat-a-time method and Optimization was performed by response surface methodology. MEGA 6 software was used for phylogenetic analyses. To analyze the data, the Design Expert 7 software and the one-way analysis of variance were used. Findings The maximum protease production, which was 1. 85 times higher than that of OFAT method and 3. 45 times higher than unoptimized conditions was obtained, using 1% w/v xylose, 3% w/v beef extract, 4% v/v inoculation size, pH 10, and 30° C. The established quadratic model had a great ability to predict responses to new observations due to a high value of the predicted determination coefficient. Conclusion OFAT and RSM strategies are useful screening and Optimization methods, respectively and sub I and sub II genes (alkaline serine protease genes) are detected in Bacillus pseudofirmus MSB22.

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APA: Copy

Borhani, M.S., ETEMADIFAR, Z., EMTIAZI, G., & JORJANI, E.. (2018). Molecular Detection and Optimization of Extracellular Alkaline Protease Production of Bacillus pseudofirmus MSB22 Using Response Surface Methodology. JOURNAL OF BIOTECHNOLOGY, 9(4 ), 507-515. SID. https://sid.ir/paper/231309/en

Vancouver: Copy

Borhani M.S., ETEMADIFAR Z., EMTIAZI G., JORJANI E.. Molecular Detection and Optimization of Extracellular Alkaline Protease Production of Bacillus pseudofirmus MSB22 Using Response Surface Methodology. JOURNAL OF BIOTECHNOLOGY[Internet]. 2018;9(4 ):507-515. Available from: https://sid.ir/paper/231309/en

IEEE: Copy

M.S. Borhani, Z. ETEMADIFAR, G. EMTIAZI, and E. JORJANI, “Molecular Detection and Optimization of Extracellular Alkaline Protease Production of Bacillus pseudofirmus MSB22 Using Response Surface Methodology,” JOURNAL OF BIOTECHNOLOGY, vol. 9, no. 4 , pp. 507–515, 2018, [Online]. Available: https://sid.ir/paper/231309/en

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