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 Lettjournal_title
Biotechnology lettersauthors
Markošová K,Camattari A,Rosenberg M,Glieder A,Turner NJ,Rebroš Mdoi
10.1007/s10529-017-2450-ysubject
Has Abstractpub_date
2018-01-01 00:00:00pages
127-133issue
1eissn
0141-5492issn
1573-6776pii
10.1007/s10529-017-2450-yjournal_volume
40pub_type
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