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Alternative Heterologous Expression of l-Arabinose Isomerase from Enterococcus faecium DBFIQ E36 By Residual Whey Lactose Induction

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Abstract

This study reports an alternative strategy for the expression of a recombinant l-AI from Enterococcus faecium DBFIQ E36 by auto-induction using glucose and glycerol as carbon sources and residual whey lactose as inducer agent. Commercial lactose and isopropyl β-d-1-thiogalactopyranoside (IPTG) were also evaluated as inducers for comparison of enzyme expression levels. The enzymatic extracts were purified by affinity chromatography, characterized, and applied in the bioconversion of d-galactose into d-tagatose. l-AI presented a catalytic activity of 1.67 ± 0.14, 1.52 ± 0.01, and 0.7 ± 0.04 U/mL, when expressed using commercial lactose, lactose from whey, and IPTG, respectively. Higher activities could be obtained by changing the protocol of enzyme extraction and, for instance, the enzymatic extract produced with whey presented a catalytic activity of 3.8 U/mL. The specific activity of the enzyme extracts produced using lactose (commercial or residual whey) after enzyme purification was also higher when compared to the enzyme expressed with IPTG. Best results were achieved when enzyme expression was conducted using 4 g/L of residual whey lactose for 11 h. These results proved the efficacy of an alternative and economic protocol for the effective expression of a recombinant l-AI aiming its high-scale production.

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Acknowledgements

The authors acknowledge the Brazilian Research Agencies CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), and FUNCAP (Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico) for the financial support.

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Authors and Affiliations

  1. Departamento de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, Bloco 709, Fortaleza, CE, CEP 60455-760, Brazil

    Ticiane C. de Souza, Ravena Casemiro Oliveira & Luciana R. B. Gonçalves

  2. Departamento de Biologia, Universidade Federal do Ceará, Campus do Pici, Bloco 909, Fortaleza, CE, CEP 60440-900, Brazil

    Saulo Gonçalves Santiago Bezerra & Denise Cavalcante Hissa

  3. Grupo de Ingeniería de Alimentos y Biotecnología, Instituto de Desarrollo Tecnológico para la Industria Química, Universidad Nacional del Litoral, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), RN 168 Km 472 “Paraje El Pozo” S/N, Santa Fe, Argentina

    Ricardo M. Manzo & Enrique J. Mammarella

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  1. Ticiane C. de Souza
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Correspondence to Denise Cavalcante Hissa or Luciana R. B. Gonçalves.

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de Souza, T.C., Oliveira, R.C., Bezerra, S.G.S. et al. Alternative Heterologous Expression of l-Arabinose Isomerase from Enterococcus faecium DBFIQ E36 By Residual Whey Lactose Induction. Mol Biotechnol 63, 289–304 (2021). https://doi.org/10.1007/s12033-021-00301-2

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