Background: The genus Thermus, which has been considered for a long time as a fruitful source of biotechnological relevant enzymes, has emerged more recently as suitable host to overproduce thermozymes. Among these, α-galactosidases are widely used in several industrial bioprocesses that require high working temperatures and for which thermostable variants offer considerable advantages over their thermolabile counterparts. Results: Thermus thermophilus HB27 strain was used for the homologous expression of the TTP0072 gene encoding for an α-galactosidase (TtGalA). Interestingly, a soluble and active histidine-tagged enzyme was produced in larger amounts (5 mg/L) in this thermophilic host than in Escherichia coli (0.5 mg/L). The purified recombinant enzyme showed an optimal activity at 90 °C and retained more than 40% of activity over a broad range of pH (from 5 to 8). Conclusions: TtGalA is among the most thermoactive and thermostable α-galactosidases discovered so far, thus pointing to T. thermophilus as cell factory for the recombinant production of biocatalysts active at temperature values over 90 °C.

Thermus thermophilus as source of thermozymes for biotechnological applications: homologous expression and biochemical characterization of an α‑galactosidase / Aulitto, Martina; Fusco, Salvatore; Fiorentino, Gabriella; Limauro, Danila; Pedone, EMILIA MARIA; Bartolucci, Simonetta; Contursi, Patrizia. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - 16:28(2017). [doi: 10.1186/s12934-017-0638-4]

Thermus thermophilus as source of thermozymes for biotechnological applications: homologous expression and biochemical characterization of an α‑galactosidase

AULITTO, MARTINA;FIORENTINO, GABRIELLA;LIMAURO, DANILA;PEDONE, EMILIA MARIA;BARTOLUCCI, SIMONETTA;CONTURSI, PATRIZIA
2017

Abstract

Background: The genus Thermus, which has been considered for a long time as a fruitful source of biotechnological relevant enzymes, has emerged more recently as suitable host to overproduce thermozymes. Among these, α-galactosidases are widely used in several industrial bioprocesses that require high working temperatures and for which thermostable variants offer considerable advantages over their thermolabile counterparts. Results: Thermus thermophilus HB27 strain was used for the homologous expression of the TTP0072 gene encoding for an α-galactosidase (TtGalA). Interestingly, a soluble and active histidine-tagged enzyme was produced in larger amounts (5 mg/L) in this thermophilic host than in Escherichia coli (0.5 mg/L). The purified recombinant enzyme showed an optimal activity at 90 °C and retained more than 40% of activity over a broad range of pH (from 5 to 8). Conclusions: TtGalA is among the most thermoactive and thermostable α-galactosidases discovered so far, thus pointing to T. thermophilus as cell factory for the recombinant production of biocatalysts active at temperature values over 90 °C.
2017
Thermus thermophilus as source of thermozymes for biotechnological applications: homologous expression and biochemical characterization of an α‑galactosidase / Aulitto, Martina; Fusco, Salvatore; Fiorentino, Gabriella; Limauro, Danila; Pedone, EMILIA MARIA; Bartolucci, Simonetta; Contursi, Patrizia. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - 16:28(2017). [doi: 10.1186/s12934-017-0638-4]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/666049
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