Response Surface Methodology for Optimization Membrane Disruption Using Thermolysis in Lipase Lk2 and Lk3

Lk2 and Lk3 were thermostable recombinant lipase and highly expressed in Escherichia coli BL21 (DE3). However, Lk2 and Lk3 accumulated as an inclusion body. To further characterize both recombinant lipases, the soluble enzyme must be obtained first. This study aimed to optimize the disruption of the...

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Bibliographic Details
Published in:Journal of pure & applied microbiology : an international research journal of microbiology 2022-06, Vol.16 (2), p.1274-1283
Main Authors: Haryati, Titin, Widhiastuty, Made Puspasari, Warganegara, Fida Madayanti
Format: Article
Language:English
Subjects:
Analysis
Detergents, Synthetic
Escherichia coli
inclusion body
Lipase
Methods
Pyrolysis
recombinant
thermolysis
thermostable
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Summary:Lk2 and Lk3 were thermostable recombinant lipase and highly expressed in Escherichia coli BL21 (DE3). However, Lk2 and Lk3 accumulated as an inclusion body. To further characterize both recombinant lipases, the soluble enzyme must be obtained first. This study aimed to optimize the disruption of the cell membrane in order to obtain soluble and active lipases. The effects of temperature lysis, pH, and SDS concentration on lipolytic activity Lk2 and Lk3 were investigated using a three-factor Box-Behnken design response surface methods. The optimum condition for the temperature variables at 50°C, pH 8, and 0.34% SDS which gave a lipolytic activity of 0.9 U for Lk2. Meanwhile, Lk3 lipolytic activity of 0.9 U obtained at the temperature of 50°C, pH 8, and 0.1% SDS. This result showed efficient one-step membrane disruption methods using thermolysis with addition of a low concentration of detergent at pH 8. The methods used were effective and applicable in the production of active and soluble thermostable recombinant lipase.
ISSN:0973-7510
2581-690X
DOI:10.22207/JPAM.16.2.56