Heterologous expression of a thermophilic esterase in Kluyveromyces yeasts

Autor(es)

Rocha, Saul Nitsche ^[1] ; Abrahao-Neto, Jose ^[2] ; Cerdan, Maria Esperanza ^[3] ; Gombert, Andreas Karoly ^[1] ; Gonzalez-Siso, Maria Isabel ^[3]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Chem Engn, BR-05424970 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Sch Pharmaceut Sci, BR-05508900 Sao Paulo - Brazil

^[3] Univ A Coruna, Fac Ciencias, Dept Biol Celular & Mol, La Coruna 15071 - Spain

• Tipo de documento: Artigo Científico
• Fonte: Applied Microbiology and Biotechnology; v. 89, n. 2, p. 375-385, JAN 2011.
• Citações Web of Science: 21
• Assunto(s): Kluyveromyces Saccharomyces cerevisiae Fermentação alcoólica

Resumo

In the present work, a thermophilic esterase from Thermus thermophilus HB27 was cloned into Kluyveromyces marxianus and into Kluyveromyces lactis using two different expression systems, yielding four recombinant strains. K. lactis showed the highest esterase expression levels (294 units per gram dry cell weight, with 65% of cell-bound enzyme) using an episomal system with the PGK promoter and terminator from Saccharomyces cerevisiae combined with the K. lactis k1 secretion signal. K. marxianus showed higher secretion efficiency of the heterologous esterase (56.9 units per gram dry cell weight, with 34% of cell-bound enzyme) than K. lactis. Hydrolytic activities for the heterologous esterases were maximum at pH values between 8.0 and 9.0 for both yeast species and at temperatures of 50 A degrees C and 45 A degrees C for K. marxianus and K. lactis, respectively. When compared to previously published data on this same esterase produced in the original host or in S. cerevisiae, our results indicate that Kluyveromyces yeasts can be considered good hosts for the heterologous secretion of thermophilic esterases, which have a potential application in biodiesel production or in resolving racemates. (AU)

• Processo FAPESP: 05/55835-3 - Desenvolvimento de um sistema de expressão e excreção de proteínas heterólogas na levedura Kluyveromyces marxianus
• Beneficiário: Saul Nitsche Rocha
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 01/13615-6 - Caracterização fisiológica da levedura Kluyveromyces marxianus visando sua utilização como fábrica celular
• Beneficiário: Andreas Karoly Gombert
• Modalidade de apoio: Auxílio à Pesquisa - Regular