Statistical Optimization and Secretion of NaCl-Tolerant Proteinases by a Moderate Halophile, Virgibacillus sp. SK37

Sinuwan, Sornchai; Jangchud, Anuvat; Rodtong, Sureelak; Roytrakul, Sittirak; Yongsawatdigul, Jirawat

doi:10.17113/ftb.53.02.15.4015

Cited by 3 publications

(2 citation statements)

References 23 publications

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“…The presence of the divalent calcium ion has been found to enhance the secretion of the protease in the production medium (30). In a few cases, the medium containing natural substrates such as fishbased medium (31) or vegetable wastes (32) have also been considered for optimizing production of the proteases. The production/activity of the most of these serine proteases was found to be dependent on the concentration of the divalent calcium ions.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Parameters that Affect the Activity of the Alkaline Protease from Halotolerant Bacterium, Bacillus acquimaris VITP4, by the Application of Response Surface Methodology and Evaluation of the Storage Stability of the Enzyme

2016

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A great deal of efforts is focused on meeting the increasing demand for novel microbial catalysts that are capable of functioning under extreme conditions, and therefore, the production of these novel microbial enzymes with appreciable activity and stability in the harsh environments (1). Enzymes from organisms which can grow in 0-15% of the NaCl will have great potential for the use in the industries because of their inherent ability to be active and stable both in the presence as well as in the absence of salt. However, most of these enzymes require metal ions for the maintenance of the structure and/or stability (2). These enzymes are active and stable under conditions that cause in a lower activity, which include high salt concentrations or organic solvents (3). In most of the cases, measurable proteolytic activity is achieved upon supplementation of cofactors like divalent metal ions in the assaying buffer. Also loss of enzymatic activity can be restored by incubating the enzyme with excess metal ions (4). Background: It was previously shown that the activity of a serine protease from a moderately halotolerant Bacillus aquimaris VITP4 strain is active in a wide range of pH and temperatures and could be modulated by the presence of the divalent metal ions. Objectives: In the present study, a quantitative analysis was done in order to explore the parameters that are contributing to the protease activity. Materials and Methods: Changes in the secondary structure of the enzyme was determined by circular dichroism analysis. The conditions for the optimal activity was investigated by Response Surface Methodology. Stability of the enzyme was determined by thermal inactivation experiments. Results:The initial one-factor-at-a-time experiments have indicated that the activity of the enzyme could be enhanced not only by the presence of low concentrations of NaCl but also by divalent metal ions, such as Ca 2+ , Mn 2+ and Cu 2+ . A clear dependence of the activity to the secondary structure of the enzyme could be established using circular dichroism spectroscopy. In the next level of optimization, four factors; viz. pH, temperature, concentration of Ca 2+ , and Mn 2+ were used to optimize the conditions required for the maximal activity of the enzyme by Response Surface Methodology, and the data could be explained using quadratic model. Under optimal condition of 43°C, pH 8.0, 8.2 mM Ca 2+ , and 4.3 mM Mn 2+ a 1.5 times enhancement in the enzyme activity could be achieved. The storage stability of the enzyme under these selected conditions has indicated a non-linear relation between the conditions for the enzymatic activity as well as stability. However, the condition for the maximal stability (267±18 min) has corresponded to that of the optimal conditions for the maximal activity. Conclusions: This study, for the first time, has explored the possibility of using statistical methods for identifying the optimal conditions for alkaline protease activity isolated from the halotolerant Bacillus aquimaris VITP4.

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Section: Discussionmentioning

confidence: 99%

Optimization of Parameters that Affect the Activity of the Alkaline Protease from Halotolerant Bacterium, Bacillus acquimaris VITP4, by the Application of Response Surface Methodology and Evaluation of the Storage Stability of the Enzyme

2016

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“…Virgibacillus sp. was also studied for NaCl-tolerant protease production by optimizing the salt concentration in the medium using RSM [34]. Saxena & Singh [35] performed two step statistical approach for the simultaneous production of amylase and protease by Solid state fermentation via Bacillus megaterium.…”

Section: Introductionmentioning

confidence: 99%

Medium Optimization for Protease Production via Moderately Halotolerant Virgibacillus Pantothenticus Using Response Surface Methodology

Banger

Nikerel

2020

Eskişehir Teknik Üniversitesi Bilim Ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji

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Virgibacillus pantothenticus is an industrially promising, yet scarcely studied, moderately halotolerant microorganism (up to 10% NaCl) with high activity protease production potential. Following Response Surface Methodology, we employed a Central Composite Design for the experiments and constructed a second order polynomial model to represent the resulting data. For medium optimization for protease production we optimized the resulting model. 32 experiments (following the central composite design scheme) where, carbon (glucose), nitrogen (ammonium sulfate), potassium (potassium phosphate monobasic) and magnesium (magnesium sulfate) sources were studied in the media. Bacterial growth, residual glucose and protease activities were determined at the 48 th hour of each experiment. The model was optimized and the concentrations were found for each parameter. Under the optimum conditions, the predicted protease activity is also experimentally verified and the model prediction was in very good agreement with experimental results. The interactions between medium components and their effect on cell growth and protease production are also sought. This work reports the improvements on protease production of a potentially interesting industrial host, Virgibacillus pantothenticus.

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Low-cost culture medium for the production of proteases by Bacillus mojavensis SA and their potential use for the preparation of antioxidant protein hydrolysate from meat sausage by-products

2018

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Statistical Optimization and Secretion of NaCl-Tolerant Proteinases by a Moderate Halophile, Virgibacillus sp. SK37

Cited by 3 publications

References 23 publications

Optimization of Parameters that Affect the Activity of the Alkaline Protease from Halotolerant Bacterium, Bacillus acquimaris VITP4, by the Application of Response Surface Methodology and Evaluation of the Storage Stability of the Enzyme

Optimization of Parameters that Affect the Activity of the Alkaline Protease from Halotolerant Bacterium, Bacillus acquimaris VITP4, by the Application of Response Surface Methodology and Evaluation of the Storage Stability of the Enzyme

Medium Optimization for Protease Production via Moderately Halotolerant Virgibacillus Pantothenticus Using Response Surface Methodology

Low-cost culture medium for the production of proteases by Bacillus mojavensis SA and their potential use for the preparation of antioxidant protein hydrolysate from meat sausage by-products

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