A comparative study of maltooligosyltrehalose synthase from Sulfolobus acidocaldarius expressed in Pichia pastoris and Escherichia coli. (September 2017)

• Record Type: Journal Article
• Authors: Han, Chang; Su, Lingqia; Hong, Ruoyu; Wu, Shixiong; Wu, Jing
• Is Part Of: Process biochemistry. Volume 60(2017)
• Page Start: 35
• Page End: 41
• Publication Date: 2017-09
• Subjects: Maltooligosyltrehalose synthase -- Trehalose -- Escherichia coli -- Pichia pastoris -- Sulfolobus acidocaldarius; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2017.05.025
• Languages: English
• ISSNs: 1359-5113

Abstract

Highlights: This was the first time that the gene treY of MTSase was expressed in Pichia pastoris. . In this paper, the highest level of thermostable MTSase expression was achieved. Abstract: Two genes encoding the maltooligosyltrehalose synthase (MTSase) from Sulfolobus acidocaldarius ATCC 33909 were synthesized: one (designated E- treY ) using codons preferred by Escherichia coli and the other (designated P- treY ) using codons preferred by Pichia pastoris . E- treY was inserted into pET-24a(+), and the resulting plasmid was used to transform E. coli BL21(DE3). P- treY was inserted into pPIC3.5 K and pGAPZA, and the resulting plasmids were used to transform P. pastoris KM71. The maximum MTSase activity expressed by these E. coli and P. pastoris strains reached 31.6 and 133.0 U mL −1, respectively. The purified MTSases displayed the same pH (pH 5.5) and temperature (75 °C) optima. Both had a half-life of 12 h at 75 °C. The recombinant P. pastoris KM71/pPIC3.5K- treY strain producing the highest enzyme activity was used to optimize the fermentation conditions in a 3.6-L bioreactor. Fermentation for 96 h under optimal conditions yielded an MTSase activity of 747.4 U mL −1 .