Immobilization of Bacillus megaterium MTCC 2444 by Ca-alginate entrapment method for enhanced alkaline protease production

This research aims to produce thermostable alkaline protease from Bacillus subtilis isolated from La Hot Spring, Terengganu, Malaysia. The study was also conducted to determine the optimum conditions for protease production and stability by considering several parameters including pH, temperature and salt concentration. All seven bacteria were screened on skim milk agar overnight at 37 °C. Three strains with the highest proteolytic activity were identified in protease specific medium. The thermostable alkaline protease had an optimum temperature of 60 °C which achieved 85.73, 82.90 and 83.05 U/mL of protease activity for the three strains respectively. Furthermore, the strains exhibited significant activity of more than 90% from their original activity. Meanwhile, the optimum pH for protease production was pH 9 with the protease activity of 76.76, 79.71 and 88.39 U/mL for TB4, TB6 and TB9 strains, respectively. Proteases were found stable at pH 9 where the loss did not exceed 30% of its original activity. Collectively, all of the data emphasised that proteases from B. subtilis were alkaline thermostable proteases in accordance with a recent report. The finding highlights the viability of the proteases for biotechnological and industrial applications.

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Application of statistical experimental design for optimization of alkaline protease production from Bacillus sp. RGR-14

Process Biochemistry ◽

10.1016/j.procbio.2003.11.002 ◽

2004 ◽

Vol 39 (12) ◽

pp. 2115-2122 ◽

Cited By ~ 132

Author(s):

Bhavna Chauhan ◽

Rani Gupta

Keyword(s):

Experimental Design ◽

Alkaline Protease ◽

Protease Production ◽

Statistical Experimental Design ◽

Bacillus Sp

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Characterization of alkaline protease produced by Bacillus megaterium

Journal of Biotechnology ◽

10.1016/j.jbiotec.2017.06.1013 ◽

2017 ◽

Vol 256 ◽

pp. S63

Author(s):

Melis Sumengen Ozdenefe ◽

Sadik Dincer ◽

Mustafa Umit Unal ◽

Hatice Aysun Mercimek Takci ◽

Fikret Buyukkaya Kayis ◽

...

Keyword(s):

Bacillus Megaterium ◽

Alkaline Protease

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Alkaline protease production from alkalophilic Bacillus sp. isolated from natural habitats

Enzyme and Microbial Technology ◽

10.1016/j.enzmictec.2005.12.004 ◽

2006 ◽

Vol 39 (4) ◽

pp. 703-710 ◽

Cited By ~ 61

Author(s):

H. Genckal ◽

C. Tari

Keyword(s):

Alkaline Protease ◽

Protease Production ◽

Bacillus Sp ◽

Natural Habitats ◽

Alkalophilic Bacillus

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OPTIMIZATION OF ALKALINE PROTEASE PRODUCTION BY STREPTOMYCES AMBOFACIENS IN FREE AND IMMOBILIZED FORM

American Journal of Biochemistry and Biotechnology ◽

10.3844/ajbbsp.2014.1.13 ◽

2014 ◽

Vol 10 (1) ◽

pp. 1-13 ◽

Cited By ~ 12

Author(s):

Abdelwahed

Keyword(s):

Alkaline Protease ◽

Protease Production ◽

Streptomyces Ambofaciens

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Statistical Approach for Optimization of Physiochemical Requirements on Alkaline Protease Production from Bacillus licheniformis NCIM 2042

Enzyme Research ◽

10.1155/2012/905804 ◽

2012 ◽

Vol 2012 ◽

pp. 1-13 ◽

Cited By ~ 26

Author(s):

Biswanath Bhunia ◽

Apurba Dey

Keyword(s):

Bacillus Licheniformis ◽

Alkaline Protease ◽

Tween 80 ◽

Culture Conditions ◽

Protease Production ◽

Sodium Perborate ◽

Physiochemical Parameters ◽

Peg 4000 ◽

Detergent Stability ◽

Triton X

The optimization of physiochemical parameters for alkaline protease production using Bacillus licheniformis NCIM 2042 were carried out by Plackett-Burman design and response surface methodology (RSM). The model was validated experimentally and the maximum protease production was found 315.28 U using optimum culture conditions. The protease was purified using ammonium sulphate (60%) precipitation technique. The HPLC analysis of dialyzed sample showed that the retention time is 1.84 min with 73.5% purity. This enzyme retained more than 92% of its initial activity after preincubation for 30 min at 37∘C in the presence of 25% v/v DMSO, methanol, ethanol, ACN, 2-propanol, benzene, toluene, and hexane. In addition, partially purified enzyme showed remarkable stability for 60 min at room temperature, in the presence of anionic detergent (Tween-80 and Triton X-100), surfactant (SDS), bleaching agent (sodium perborate and hydrogen peroxide), and anti-redeposition agents (Na2CMC, Na2CO3). Purified enzyme containing 10% w/v PEG 4000 showed better thermal, surfactant, and local detergent stability.

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