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Updated: Feb 21, 2026
Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies
Published on: January 23, 2019
Cloning and upscale production of monoamine oxidase N (MAO-N D5) by Pichia pastoris
Kristína Markošová1, Andrea Camattari2,3, Michal Rosenberg1
1Faculty of Chemical and Food Technology, Institute of Biotechnology, Slovak University of Technology, Radlinského 9, 812 37, Bratislava, Slovakia.
Objective:
To clone monoamine oxidase N, that catalyses the selective oxidative deamination or deracemisation of amines into imines, in Pichia pastoris and prove the importance of choosing the proper expression system for its recombinant production.
Results:
Monoamine oxidase, originating from Aspergillus niger and subjected to directed evolution (MAO-N D5), was cloned and expressed in Pichia pastoris CBS7435 MutS strain for the first time. Various transformants were screened at microscale level. The production of the clone expressing the most active enzyme was scaled-up to a 1.5 l fermenter and preparation of MAO-N D5 as a crude enzyme extract was optimised. The obstacles in the production of the enzyme in both expression systems, Escherichia coli and P. pastoris, are discussed and demonstrated.
Conclusions:
There was an improvement in specific productivity, which was 83 times higher in P. pastoris, clearly proving the importance of choosing the right expression host system for the specific enzymes.
