The <i>Pichia pastoris</i> enzyme production platform: From combinatorial library screening to bench-top fermentation on residual cyanobacterial biomass

Korbinian Sinzinger; Ulrike Obst; Samed Güner; Manuel Döring; Magdalena Haslbeck; Doris Schieder; Volker Sieber

doi:10.1016/j.jobab.2023.12.005

Volume 9 Issue 1

Feb. 2024

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Korbinian Sinzinger, Ulrike Obst, Samed Güner, Manuel Döring, Magdalena Haslbeck, Doris Schieder, Volker Sieber. The Pichia pastoris enzyme production platform: From combinatorial library screening to bench-top fermentation on residual cyanobacterial biomass[J]. Journal of Bioresources and Bioproducts, 2024, 9(1): 43-57. doi: 10.1016/j.jobab.2023.12.005

Citation:

Korbinian Sinzinger, Ulrike Obst, Samed Güner, Manuel Döring, Magdalena Haslbeck, Doris Schieder, Volker Sieber. The Pichia pastoris enzyme production platform: From combinatorial library screening to bench-top fermentation on residual cyanobacterial biomass[J]. Journal of Bioresources and Bioproducts, 2024, 9(1): 43-57. doi: 10.1016/j.jobab.2023.12.005

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The Pichia pastoris enzyme production platform: From combinatorial library screening to bench-top fermentation on residual cyanobacterial biomass

doi: 10.1016/j.jobab.2023.12.005

a. Chair of Chemistry of Biogenic Resources, Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Straubing 94315, Germany;
b. Catalysis Research Center, Technical University of Munich, Garching 85748, Germany;
c. SynBiofoundry@TUM, Technical University of Munich, Straubing 94315, Germany;
d. School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia 4072, Australia

Funds:

The EFRE—Interreg project 41 “Joint Research on Natural Compounds from Cyanobacteria as a Model of Cross-Border Scientific Partnership” was funded by the European Union ZIEL ETZ. Many thanks to Algatech for providing the biomass and to Prof. Timothy K. Lu (MIT) for providing P. pastoris attP.

Available Online: 2024-01-31
Publish Date: 2023-12-26

Abstract

Abstract

The demand for industrial enzymes is continually rising, fueled by the growing need to shift towards more sustainable industrial processes. However, making efficient enzyme production strains and identifying optimal enzyme expression conditions remains a challenge. Moreover, the production of the enzymes themselves comes with unavoidable impacts, e.g., the need to utilize secondary feedstocks. Here, we take a more holistic view of bioprocess development and report an integrative approach that allows us to rapidly identify improved enzyme expression and secretion conditions and make use of cyanobacterial waste biomass as feed for supporting Pichia pastoris fermentation. We demonstrate these capabilities by producing a phytase secreted by P. pastoris that is grown on cyanobacterium hydrolysate and buffered glycerol-complex (BMGY) medium, with genetic expression conditions identified by high-throughput screening of a randomized secretion library. When our best-performing strain is grown in a fed-batch fermentation on BMGY, we reach over 7 000 U/mL in three days.
- Protein secretion,
- Phytase,
- Cyanobacterial biomass,
- Fed-batch fermentation,
- Pichia pastoris

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References(47)

References

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