Cloning and upscale production of monoamine oxidase N (MAO-N D5) by Pichia pastoris.

Abstract:

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.

journal_name

Biotechnol Lett

journal_title

Biotechnology letters

authors

Markošová K,Camattari A,Rosenberg M,Glieder A,Turner NJ,Rebroš M

doi

10.1007/s10529-017-2450-y

subject

Has Abstract

pub_date

2018-01-01 00:00:00

pages

127-133

issue

1

eissn

0141-5492

issn

1573-6776

pii

10.1007/s10529-017-2450-y

journal_volume

40

pub_type

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