Expression of xyloglucan endotransglycosylases of Gerbera hybrida and Betula pendula in Pichia pastoris

Autor: Juha Immanen, Teemu H. Teeri, Michael Bailey, Paula Elomaa, Eija Rintala, Yrjö Helariutta, Jaana Toikkanen, Marja-Leena Niku-Paavola, Richard Fagerström
Rok vydání: 2006
Předmět:
0106 biological sciences
Gerbera
Bioreactor cultivation
Gerbera hybrida
Molecular Sequence Data
Cell Culture Techniques
Gene Expression
Xyloglucan endotransglycosylase (XET)
Bioengineering
Asteraceae
01 natural sciences
Applied Microbiology and Biotechnology
Pichia
Pichia pastoris
Cell wall
03 medical and health sciences
chemistry.chemical_compound
Bioreactors
Botany
Amino Acid Sequence
Biomass
Betula
030304 developmental biology
2. Zero hunger
chemistry.chemical_classification
0303 health sciences
biology
Molecular mass
Sequence Homology
Amino Acid
Methanol
Temperature
Glycosyltransferases
General Medicine
biology.organism_classification
Yeast
Culture Media
Xyloglucan
Enzyme
chemistry
Biochemistry
Betula pendula
Heterologous expression
Filtration
010606 plant biology & botany
Biotechnology
Zdroj: Toikkanen, J H, Niku-Paavola, M-L, Bailey, M, Immanen, J, Rintala, E, Elomaa, P, Helariutta, Y, Teeri, T H & Fagerström, R 2007, ' Expression of xyloglucan endotransglycosylases of Gerbera hybrida and Betula pendula in Pichia pastoris ', Journal of Biotechnology, vol. 130, no. 2, pp. 161-170 . https://doi.org/10.1016/j.jbiotec.2007.03.004
ISSN: 0168-1656
Popis: The plant enzyme xyloglucan endotransglycosylase (XET; EC 2.4.1.207, xyloglucan:xyloglucosyl transferase) participates in selective modification of plant cell walls during cell growth. XETs are potential catalysts in various applications. Here, sequences encoding two XETs from Gerbera hybrida and Betula pendula are reported. The encoded proteins, which are 51% identical at the amino acid level, were expressed in the yeast Pichia pastoris in secreted form with the aid of mating factor alpha signal sequence. XET production in shake flask cultivations was better at 22 °C than at 30 °C. Both the yield of protein of expected molecular mass and the XET activity improved at the lower temperature. Under all cultivation conditions studied, higher amounts of XET from B. pendula (BXET) were expressed than XET from G. hybrida (GXET). Both XET enzymes were produced in 16 l fed-batch bioreactor cultures. GXET was produced in methanol-limited fed-batch cultivation in minimal medium, and BXET in temperature-limited fed-batch (TLFB) in minimal or complex medium. Production was highest in TLFB in complex medium. BXET was purified from the culture filtrate and characterized. Based on the specific activity of the purified protein, 60–70 mg l−1 BXET was produced in the TLFB in complex medium.
Databáze: OpenAIRE
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