CD Laboratory for Growth-Decoupled Protein Production in Yeast
- Lebensmittel, Ernährung, Gesundheit
- Biotechnologie
Abstract
Yeasts such as Pichia pastoris are commonly used as platform for production of recombinant proteins. Commonly, high biomass concentrations are used during production processes due to two reasons: 1) the low specific productivity is compensated by high biomass concentration and 2) protein production is correlated with growth, so that continuous biomass accumulation is needed for efficient product formation. This creates several problems in large scale production, such as high oxygen demand and heat generation (requiring high aeration and cooling). High cell density also causes problems during cell separation, especially for secreted products the removal of cells from the supernatant is a technical challenge. Furthermore, the use of disposables with microbial systems such as P. pastoris is presently limited due to the high biomass concentrations. Therefore, this project aims at developing a new P. pastoris expression platform that maintains high productivity at low growth rates by cell engineering at several levels. The scientific challenge is to uncouple protein synthesis and secretion from growth. Therefore cell metabolism shall be studied at (near) zero growth rates (in retentostats) to enable better understanding of the metabolic and cellular limitations at such slow growth rates. Cell engineering will then tackle these limitations. Together, both strategies lead to the design of optimal processes for high productivity of secreted recombinant proteins.
Publications
Engineering transcription regulation in Pichia pastoris.
Autoren: Flores Villegas, MC; Mattanovich, D; Gasser, B Jahr: 2018
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Regulation of pseudohyphal growth and flocculation in Pichia pastoris
Autoren: De, S; Rebnegger, C; Moser, J; Tatto, N.E; Mattanovich, D; Gasser, B; Jahr: 2018
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Characterization of transcription factors in Pichia pastoris
Autoren: Flores Villegas, M; Mattanovich, D; Gasser, B Jahr: 2019
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Preventing morphological cell heterogeneity in the yeast Pichia pastoris (syn Komagataella phaffii): involvement of the FLO gene family.
Autoren: De, S; Rebnegger, C; Buchetics, M; Moser, J; Tatto, N; Graf, AB; Mattanovich, D; Gasser, B. Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Pichia production host engineering by systemic host changes
Autoren: Kiziak, C; Adelanto, N; Staudacher, J; Rebnegger, C; Gassser, B; Mattanovich, D Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Investigating the role of oxidative stress response genes in protein secretion using a novel combinatorial Golden Gate based approach
Autoren: Mendes Honorato, V; Staudacher, J; Prielhofer, R; Gasser, B Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Treatment with surfactants enables quantification of translational activity by O-propargyl-puromycin labelling in yeast
Autoren: Staudacher, J; Rebnegger, C; Gasser, B Jahr: 2021
Journal articles
Investigating the role of oxidative stress response genes in protein secretion using a novel Golden PiCS-based approach.
Autoren: Mendes Honorato, V; Staudacher, J; Gasser B. Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Unfolded protein response biosensors for recombinant protein expression
Autoren: Godon, H; Puricelli, S; De, S; Adalanto, C; Kiziak, C; Gasser, B; Mattanovich, D Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Going beyond the limit: impact of increasing global translation activity on the productivity of recombinant secreted proteins in Pichia pastoris
Autoren: Staudacher, J; Rebnegger, C; Mattanovich, D; Gasser, B Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Maintenance energy requirement largely determines metabolic rewiring of Komagataella phaffii at glucose limited near zero growth
Autoren: Coltman, B; Rebnegger, C; Gasser, B; Zanghellini, J Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:In pursuit of an orthogonal molecular switch for Komagataella phaffii
Autoren: Malecki, P; Rebnegger, C; Gasser, B Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Maintenance energy requirements determine metabolic rewiring of Komagataella phaffii at glucose-limited near-zero growth conditions
Autoren: Coltman, B; Rebnegger, C; Gasser, B; Zanghellini, J Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Gasser, B
Autoren: Pushing and pulling proteins into the yeast secretory pathway enhances recombinant protein secretion Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:In vivo monitoring of reactive oxygen species in the yeast Komagataella phaffii using a genetically encoded fluorescent H2O2 biosensor
Autoren: Honorato, VM; Molin, M; Gasser, B Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Characterising an orthogonal thiamine-responsive riboswitch for expression control in the yeast Komagataella phaffii
Autoren: Malecki, P; Rebnegger, C; Gasser B Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Protein production dynamics and physiological adaptation of recombinant Komagataella phaffii at near-zero growth rates
Autoren: Rebnegger, C; Kowarz, V; Coltman, B; Mattanovich, D; Gasser, B Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:In pursuit of an orthogonal molecular switch for Komagataella phfaffii
Autoren: Malecki, P; Rebnegger, C; Gasser, B Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:What makes Komagataella phaffii non-conventional?
Autoren: Ata, O; Ergun, BG; Fickers, P; Heistinger, L; Mattanovich, D; Rebnegger, C; Gasser, B Jahr: 2021
Journal articles
Investigating the role of oxidative stress response genes in protein secretion using a novel Golden PiCS-based approach
Autoren: Mendes Honorato, V; Staudacher, J; Gasser B Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Going beyond the limit: Increasing global translation activity leads to increased productivity of recombinant secreted proteins in Pichia pastoris
Autoren: Staudacher, J; Rebnegger, C; Dohnal, T; Landes, N; Mattanovich, D; Gasser, B Jahr: 2022
Journal articles
Treatment with surfactants enables quantification of translational activity by O-propargyl-puromycin labelling in P. pastoris.
Autoren: Staudacher, J; Rebnegger, C; Gasser, B Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Treatment with surfactants enables quantification of translational activity by O-propargyl-puromycin labelling in P. pastoris.
Autoren: Staudacher, J; Rebnegger, C; Gasser, B Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Assessing flux rearrangements of an alternate methanol metabolism that enables efficient recombinant protein production in Pichia pastoris
Autoren: Coltman, B; Zavec, D; Mattanovich, D; Gasser, B; Zanghellini, J Jahr: 2021
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Pichia production platform engineering by systemic changes
Autoren: Kiziak, C; Adelantado, N; Staudacher, J; Rebnegger, C; Gasser, B; Mattanovich, D Jahr: 2022
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Going beyond the limit: tuning global transcription and translation activity increases recombinant protein secretion in Komagataella phaffii
Autoren: Gasser, B; Staudacher, J; Rebnegger, C; Mattanovich, D Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Going beyond the limit: impact of increasing global translation activity on the productivity of recombinant secreted proteins in Pichia pastoris
Autoren: Staudacher, J; Rebnegger, C; Mattanovich, D; Gasser, B Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Increasing global translation activity and its impact on Komagataella phaffii
Autoren: Staudacher, J; Rebnegger, C; Mattanovich, D; Gasser B Jahr: 2023
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Systematic sequence engineering enhances the induction strength of the glucose-regulated GTH1 promoter of Komagataella phaffii.
Autoren: Flores-Villegas, M; Rebnegger, C; Kowarz, V; Prielhofer, R; Mattanovich, D; Gasser, B; Jahr: 2023
Journal articles
Slow Growth Rate triggered transition to a pseudohyphal lifestyle of the protein production host Pichia pastoris
Autoren: De, S; Buchetics, M; Rebnegger, C; Mattanovich, D; Gasser, B Jahr: 2019
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Dissection of growth-rate- and cell-cycle-dependent gene regulation in P-pastoris
Autoren: Rebnegger, C; Burgard, J; Graf, AB; Maurer, M; Gasser, B; Mattanovich, D Jahr: 2018
Journal articles
Protein production dynamics and physiological adaptiation of recombinant Pichia pastoris at near-zero growth rates
Autoren: Rebnegger, C; Kowarz, V; Mattanovich, D; Gasser, B Jahr: 2019
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:XS™ Pichia 2.0: closing the gap between bacterial and mammalian systems for the production of recombinant proteins
Autoren: Adelantado, N; Claes, K; Reichinger, M; Wagner, N; Prielhofer, R; Gasser, B; Mattanovich, D; Kiziak, Ch Jahr: 2019
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Impact of puromycin on different Pichia pastoris strains
Autoren: Staudacher, J: Gasser, B; Jahr: 2018
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:
Project staff
Brigitte Gasser
Assoc. Prof. Priv.Doz.DI Dr.nat.techn. Brigitte Gasser
brigitte.gasser@boku.ac.at
Tel: +43 1 47654-79033
Project Leader
01.03.2017 - 31.12.2024
Benjamin Luke Coltman
Benjamin Luke Coltman M.Sc.
benjamin.coltman@boku.ac.at
Tel: +43 1 47654-79899
Project Staff
01.01.2020 - 30.09.2022
Mirelle Citlali Flores Villegas
Mirelle Citlali Flores Villegas BSc.MSc.
mirelle.flores@boku.ac.at
Tel: +43 1 47654-79104
Project Staff
01.01.2020 - 31.03.2023
Petra Hofbauer
Mag. Petra Hofbauer MAS
petra.hofbauer@boku.ac.at
Tel: +43 1 47654-79002
Project Staff
01.03.2017 - 31.12.2024
Sharon Mano Pappu John Muthuraj
Dr. Sharon Mano Pappu John Muthuraj
Project Staff
01.02.2022 - 29.02.2024
Pawel Malecki
Mag. Pawel Malecki
pawel.malecki@boku.ac.at
Tel: +43 1 47654-79034, 79042
Project Staff
01.09.2019 - 31.12.2024
Rebecca Oberndorfer
Rebecca Oberndorfer
rebecca.oberndorfer@students.boku.ac.at
Tel: +43 1 47654-79037, 79044
Project Staff
01.01.2020 - 30.06.2020
Corinna Rebnegger
Dipl.-Ing. Corinna Rebnegger Ph.D.
corinna.rebnegger@boku.ac.at
Tel: +43 1 47654-79033, 79036
Project Staff
01.01.2020 - 29.02.2024
Jennifer Staudacher
Dipl.-Ing. Jennifer Staudacher
jennifer.staudacher@boku.ac.at
Tel: +43 1 47654-79036, 79142
Project Staff
01.01.2020 - 01.09.2022

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