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Enzymatic characterization of recombinant nitrate reductase expressed and purified from Neurospora crassa.

Fungal Genet Biol.. 2015-04;  80:10-18
Ringel P, Probst C, Dammeyer T, Buchmeier S, Jänsch L, Wissing J, Tinnefeld P, Mendel RR, Jockusch BM, Kruse T. Department of Plant Biology, Braunschweig University of Technology, 38106 Braunschweig, Germany.
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摘要

We established an expression and purification procedure for recombinant protein production in Neurospora crassa (N. crassa). This Strep-tag® based system was successfully used for purifying recombinant N. crassa nitrate reductase (NR), whose enzymatic activity was compared to recombinant N. crassa NR purified from Escherichia coli. The purity of the two different NR preparations was similar but NR purified from N. crassa showed a significantly higher nitrate turnover rate. Two phosphorylation sites were identified for NR purified from the endogenous expression system. We conclude that homologous expression of N. crassa NR yields a higher active enzyme and propose that NR phosphorylation causes enhanced enzy... More

关键词

Neurospora crassa; Nitrate reductase; Recombinant protein expression; Strep-tag®; Post-translational modification