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

Jaana H Toikkanen, Marja-Leena Niku-Paavola, Michael Bailey, Juha Immanen, Eija Rintala, Paula Elomaa, Yrjö Helariutta, Teemu H Teeri, Richard Fagerström, Teemu Teeri

Research output: Contribution to journalArticleScientificpeer-review

Abstract

"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 degrees C than at 30 degrees 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 161 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 mgl(-1) BXET was produced in the TLFB in complex medium. (c) 2007 Elsevier B.V. All rights reserved."
Original languageEnglish
JournalJournal of Biotechnology
Volume130
Pages (from-to)161-170
Number of pages10
ISSN0168-1656
DOIs
Publication statusPublished - 2007
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 414 Agricultural biotechnology
  • 411 Agriculture and forestry

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