Oxygen-limited control of methanol uptake for improved production of a single-chain antibody fragment with recombinant Pichia pastoris

Applied Microbiology and Biotechnology
Narendar K Khatri, Frank Hoffmann

Abstract

The yeast Pichia pastoris is a suitable production system for recombinant proteins due to its strong methanol-inducible AOX1 promoter. A key parameter of the production process is the specific methanol uptake rate. To control the methanol uptake and simultaneously maintain a constant methanol concentration during the production phase, two strategies were developed to generate purposeful oxygen limitation and to feed-forward control the specific methanol uptake rate into the optimum range. First, the cell density at induction was adjusted by prolonged preinduction glycerol feeding. Alternatively, the airflow rate was restricted and increased in parallel with the biomass. While the product accumulation started 20 h earlier with the first approach, the specific production rate of a single-chain antibody fragment was three times higher in the latter case. After 70 h of production, both schemes yielded product concentrations in the gram-per-liter range. Moreover, they release the requirement for dosage of pure oxygen and thereby can facilitate the scale-up of the production process. The different production profiles indicate that the impact of specific methanol uptake rate on protein production by recombinant P. pastoris depends on ...Continue Reading

References

Jul 29, 1998·Methods in Molecular Biology·J StrattonM Meagher
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Nov 21, 2000·Biotechnology and Bioengineering·M C d'Anjou, A J Daugulis
Sep 27, 2002·Current Opinion in Biotechnology·Geoff P Lin CereghinoJames M Cregg
Dec 7, 2002·Biotechnology Progress·Wenhui ZhangMichael M Meagher
Apr 4, 2003·Biotechnology and Bioengineering·M E BushellJ N Wardell
Jul 23, 2003·Microbial Cell Factories·Mehmedalija JahicSven-Olof Enfors
Feb 3, 2004·Biotechnology and Bioengineering·Hubertus HohenblumDiethard Mattanovich
Apr 28, 2004·Biotechnology and Bioengineering·A E CunhaM J T Carrondo
Aug 6, 2004·Protein Expression and Purification·Leonardo M DamascenoCarl A Batt
Dec 4, 2004·Biotechnology Progress·Oliver TrentmannFrank Hoffmann
Mar 15, 2005·Biomacromolecules·Natarajan VijayasankaranFriedrich Srienc
Aug 5, 2005·Bioprocess and Biosystems Engineering·Theppanya CharoenratSven-Olof Enfors

❮ Previous
Next ❯

Citations

Nov 23, 2006·Biotechnology Letters·Brigitte Gasser, Diethard Mattanovich
Mar 5, 2011·Microbial Cell Factories·Christian DietzschChristoph Herwig
Oct 29, 2011·Microbial Cell Factories·Christian DietzschChristoph Herwig
Sep 25, 2010·BMC Systems Biology·Moritz von StoschRui Oliveira
Jul 5, 2011·Journal of Biotechnology·Marina BerdichevskyThomas I Potgieter
Jun 12, 2010·Journal of Bioscience and Bioengineering·Gaurav BatraNavin Khanna
Dec 21, 2010·Biotechnology Journal·Ki Jun JeongNatarajan Velmurugan
May 28, 2010·Biotechnology and Bioengineering·Thomas I PotgieterMarc d'Anjou
Jun 2, 2015·Biotechnology Advances·V LooserK Kovar
Dec 15, 2007·Biotechnology and Bioengineering·Kristin BaumannDiethard Mattanovich
Feb 27, 2017·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Cesar Andres Diaz AriasAdalberto Pessoa Junior
Oct 19, 2021·Frontiers in Pharmacology·Muhammad Umair NaseemGyorgy Panyi

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.