Characterization of a Brassica napus Myrosinase Expressed and Secreted by Pichia pastoris

Frauke V. Härtel, Anders Brandt

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

In Brassica napus three different gene families with different temporal and tissue-specific expression and distribution patterns encode myrosinases (thioglucoside glucohydrolases, EC 3.2.3.1). Myrosinases encoded by the MA gene family are found as free and soluble dimers, while myrosinases encoded by the MB and MC gene families are mainly found in large insoluble complexes associated with myrosinase-binding proteins and myrosinase-associated proteins. These large complexes impede purification and characterization of MB and MC myrosinases from the plant. We used Pichia pastoris to express and secrete functional recombinant MYR1 myrosinase from B. napus to allow further characterization of myrosinase belonging to the MB gene family. The purified recombinant myrosinase hydrolyzes sinigrin with a Km of 1.0 mM; the specific activity and calculated kcat/Km were 175 U/mg and 1.9 × 105 s−1 M−1, respectively. A novel in-gel staining method for myrosinase activity is presented.
Original languageEnglish
JournalProtein Expression and Purification
Volume24
Issue number2
Pages (from-to)221-226
ISSN1046-5928
DOIs
Publication statusPublished - 2002
Externally publishedYes

Fingerprint Dive into the research topics of 'Characterization of a Brassica napus Myrosinase Expressed and Secreted by Pichia pastoris'. Together they form a unique fingerprint.

Cite this