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Production of Whole-Cell Lipase from Streptomyces clavuligerus in a Bench-Scale Bioreactor and Its First Evaluation as Biocatalyst for Synthesis in Organic Medium

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Abstract

This work evaluated a wild-type Streptomyces clavuligerus strain as a whole-cell lipase (Sc-WCL) producer by submerged fermentation. In an orbital shaker, lipase hydrolytic activity of 3000 U L−1, measured at pH 9.0 and 37 °C by using p-nitrophenyl palmitate as substrate, was achieved after 36 h fermentation using glycerol-free production medium in a baffled Erlenmeyer flask at 28 °C and pH 6.8. Maximum productivity of 52.5 U L−1 h−1 was achieved after 24 h in bioreactor using glycerol-free production medium at pH 6.8 and 28 °C, with agitation at 400 rpm and aeration at 1 vvm. Sc-WCL was shown to be more active at 60 °C and pH 10.7, while higher activity retention was observed at 30–40 °C after 1 h incubation at pH 10. Sc-WCL showed to have potential to be used as biocatalyst in hydrolysis and esterification reactions. In the hydrolysis of p-nitrophenyl palmitate, lyophilized Sc-WCL expressed a hydrolytic activity (330 units g−1 solid, measured at 37 °C and pH 9.0) around 100-fold higher than the ones declared by a supplier of lyophilized powders of mixtures of intracellular lipases from Thermus thermophiles and Thermus flavus (≥3.0 units g−1 solid, measured at 65 °C and pH 8.0). In the synthesis of butyl butyrate in anhydrous medium, 85% ester conversion was achieved at 37 °C after 8 h reaction. Thus, Sc-WCL showed to be a promising biocatalyst because it is cheaper than the isolated and purified lipases.

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Acknowledgements

The authors are grateful for the financial support provided by the Brazilian foundations CNPq (grant #142128/2012-0) and CAPES. The authors thank Prof. Alberto Colli Badino Junior (DEQ/UFSCar, São Carlos, Brazil) for the donation of the S. clavuligerus ATCC 27064 strain.

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  1. Programa de Pós-Graduação em Engenharia Química, Universidade Federal de São Carlos, Rod. Washington Luiz, km 235, São Carlos, SP, 13565-905, Brazil

    Jéssica Bravin Carmello dos Santos & Paulo Waldir Tardioli

  2. Departamento de Engenharia Química, Universidade Federal de São Carlos, Rod. Washington Luiz, km 235, São Carlos, SP, 13565-905, Brazil

    Rosineide Gomes da Silva Cruz & Paulo Waldir Tardioli

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dos Santos, J.B.C., da Silva Cruz, R.G. & Tardioli, P.W. Production of Whole-Cell Lipase from Streptomyces clavuligerus in a Bench-Scale Bioreactor and Its First Evaluation as Biocatalyst for Synthesis in Organic Medium. Appl Biochem Biotechnol 183, 218–240 (2017). https://doi.org/10.1007/s12010-017-2440-5

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