PMID- 37763289
OWN - NLM
STAT- Publisher
LR  - 20231003
IS  - 2075-1729 (Print)
IS  - 2075-1729 (Electronic)
IS  - 2075-1729 (Linking)
VI  - 13
IP  - 9
DP  - 2023 Sep 8
TI  - Optimization Workflow of Fumonisin Esterase Production for Biocatalytic 
      Degradation of Fumonisin B(1).
LID - 10.3390/life13091885 [doi]
LID - 1885
AB  - Industrial enzyme production with the Pichia pastoris expression system requires 
      a well-characterized production strain and a competitively priced fermentation 
      medium to meet the expectations of the industry. The present work shows a 
      workflow that allows the rapid and reliable screening of transformants of single 
      copy insertion of the target production cassette. A constitutive expression 
      system with the glyceraldehyde-3-phosphate dehydrogenase promoter (pGAP) with 
      homology arms for the glycerol kinase 1 (GUT1) was constructed for the targeted 
      integration of the expression plasmid in a KU70 deficient Pichia pastoris and the 
      production of a bacterial fumonisin esterase enzyme (CFE). A robust colony qPCR 
      method was developed for the copy number estimation of the expression cassette. 
      Optimization of the protein production medium and the scale-up ability was aided 
      by design of experiments (DOE) approach resulting in optimized production 
      conditions at a semi-industrial scale. A novel fermentation medium containing 3% 
      inactivated yeast and 2% dextrose in an ammonium-citrate buffer (IYD) was shown 
      to be a promising alternative to YPD media (containing yeast extract, peptone, 
      and dextrose), as similar protein titers could be obtained, while the cost of the 
      medium was reduced 20-fold. In a demonstration-scale 48 h long fed-batch 
      fermentation, the IYD media outperformed the small-scale YPD cultivation by 471.5 
      +/- 22.6%.
FAU - Incze, Daniel Janos
AU  - Incze DJ
AD  - Department of Organic Chemistry and Technology, Budapest University of Technology 
      and Economics, Muegyetem Rakpart 3, H-1111 Budapest, Hungary.
AD  - Research and Development Laboratory, Dr. Bata Ltd., Bajcsy-Zsilinszky utca 139, 
      H-2364 Ocsa, Hungary.
FAU - Poppe, Laszlo
AU  - Poppe L
AUID- ORCID: 0000-0002-8358-1378
AD  - Department of Organic Chemistry and Technology, Budapest University of Technology 
      and Economics, Muegyetem Rakpart 3, H-1111 Budapest, Hungary.
AD  - Biocatalysis and Biotransformation Research Center, Faculty of Chemistry and 
      Chemical Engineering, Babes-Bolyai University of Cluj-Napoca, Strada Arany Janos 
      11, RO-400028 Cluj-Napoca, Romania.
FAU - Bata, Zsofia
AU  - Bata Z
AUID- ORCID: 0000-0002-4840-9433
AD  - Research and Development Laboratory, Dr. Bata Ltd., Bajcsy-Zsilinszky utca 139, 
      H-2364 Ocsa, Hungary.
LA  - eng
GR  - TKP2021-EGA-02/National Research, Development and Innovation Fund (NRDIF), 
      Hungary/
GR  - KDP-2021 no. C1341189/National Research, Development, and Innovation Fund, 
      Hungary/
GR  - 2018-1.1.2-KFI-2018-00111/National Research, Development and Innovation Fund 
      (NRDIF), Hungary/
PT  - Journal Article
DEP - 20230908
PL  - Switzerland
TA  - Life (Basel)
JT  - Life (Basel, Switzerland)
JID - 101580444
PMC - PMC10533188
OTO - NOTNLM
OT  - Pichia pastoris
OT  - enzyme
OT  - fumonisin esterase
OT  - media optimization
OT  - recombinant protein production
COIS- The funders had no role in the design of the study; in the collection, analyses, 
      or interpretation of data; in the writing of the manuscript; or in the decision 
      to publish the results.
EDAT- 2023/09/28 06:42
MHDA- 2023/09/28 06:42
PMCR- 2023/09/08
CRDT- 2023/09/28 01:25
PHST- 2023/08/13 00:00 [received]
PHST- 2023/09/03 00:00 [revised]
PHST- 2023/09/06 00:00 [accepted]
PHST- 2023/09/28 06:42 [medline]
PHST- 2023/09/28 06:42 [pubmed]
PHST- 2023/09/28 01:25 [entrez]
PHST- 2023/09/08 00:00 [pmc-release]
AID - life13091885 [pii]
AID - life-13-01885 [pii]
AID - 10.3390/life13091885 [doi]
PST - epublish
SO  - Life (Basel). 2023 Sep 8;13(9):1885. doi: 10.3390/life13091885.