Production optimization, characterization, and covalent immobilization of a thermophilic Serratia rubidaea lipase isolated from an Algerian oil waste

  • Fatima Nehal
  • Mouna SahnounEmail author
  • Ahlem Dab
  • Mohammed Sebaihia
  • Samir Bejar
  • Bassem Jaouadi
Original Article


A new thermophilic non-induced lipase producer named Serratia rubidaea strain Nehal-mou was isolated from oil waste in Tissemsilat, Algeria. The most influential lipase production parameters were screened by the Plackett–Burman design for enhancing enzyme yield. An optimum condition of a 1.5% of glucose, a 0.01% of potassium, and a 0.025% of manganese contents resulted in a 41.13 U/mL. This yield was 6.29 times higher than the one achieved before the application of the Box-Behnken Design. Lipase activity showed a high organic solvent tolerance following its exposure to hexane, ethanol, methanol, and acetone. Lipase was also perfectly stable in the presence of 10 mM Fe2+, K+, and Na+ ions with more than 75% of the retaining activity. The enzyme half-life times were 22 h, 90 min, and 25 min at 50, 60, and 70 °C respectively. Polyvinyl alcohol (PVA)/boric acid/Starch/CaCO3 were utilized as a carrier for lipase covalent immobilization in order to be used efficiently. The Scanning Electron Microscopy (SEM) Technique and the Fourier Transform Infrared Spectroscopy (FTIR) Method confirmed the covalent bonding success and the excellent carrier characteristics. Thus, the immobilization yield reached 73.5% and the optimum temperature was shifted from 40 to 65 °C. The immobilized lipase kept 80% of its total activity after 10 cycles and had 3 and 3.2-fold half-lives at 70, and 80 °C respectively compared to the free enzyme.


Serratia rubidaea Lipase Thermal-stability Optimization Covalent immobilization SEM FTIR 



This research was supported by the Ministry of Higher Education and Scientific Research of  Tunisia under the contract program CBS-LMBEE/code. LR15CBS06 2015–2018 and Algeria.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Fatima Nehal
    • 1
  • Mouna Sahnoun
    • 2
    Email author
  • Ahlem Dab
    • 2
  • Mohammed Sebaihia
    • 3
  • Samir Bejar
    • 2
  • Bassem Jaouadi
    • 2
  1. 1.Faculty of Nature and Life Science, Department of Agricultural Sciences and BiotechnologiesHassiba Benbouali UniversityChlefAlgeria
  2. 2.Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS)University of SfaxSfaxTunisia
  3. 3.Laboratory of Molecular Biology, Genomics and Bioinformatics, Faculty of Nature and Life ScienceHassiba Benbouali UniversityChlefAlgeria

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