Scalable methanol‑free production of recombinant glucuronoyl esterase in Pichia pastoris
Date
2019
Journal Title
Journal ISSN
Volume Title
Publisher
BMC (part of Springer Nature)
Abstract
Objective: Glucuronoyl esterase (GE) is an emerging enzyme that improves fractionation of lignin-carbohydrate
complexes. However, the commercial availability of GE is limited, which hinders the research of GE-based bioprocesses
for its industrial application in lignocellulose biorefineries. This study evaluated a workable, cost-effective,
and commercially scalable production strategy to improve the ease of GE-based research. This strategy consisted of
a constitutive and methanol-free enzyme production step coupled with a two-step filtration process. The aim was to
determine if this strategy can yield copious amounts of GE, by secretion into the extracellular medium with an acceptable
purity that could allow its direct application. This approach was further validated for cellobiose dehydrogenase,
another emerging lignocellulose degrading enzyme which is scarcely available at high cost.
Results: The secreted recombinant enzymes were functionally produced in excess of levels previously reported for
constitutive production (1489–2780 mg L−1), and were secreted at moderate to high percentages of the total extracellular
protein (51–94%). The constant glycerol feed, implemented during fed-batch fermentation, lead to a decline
in growth rate and plateaued productivity. Tangential flow ultrafiltration was used to concentrate cell-free enzyme
extracts 5–6-fold, reaching enzyme activity levels (1020–202 U L−1) that could allow their direct application.
Description
CITATION: Conacher, C. G., et al. 2019. Scalable methanol‑free production of recombinant glucuronoyl esterase in Pichia pastoris. BMC Research Notes, 12: 596, doi:10.1186/s13104-019-4638-9.
The original publication is available at https://bmcresnotes.biomedcentral.com
Publication of this article was funded by the Stellenbosch University Open Access Fund
The original publication is available at https://bmcresnotes.biomedcentral.com
Publication of this article was funded by the Stellenbosch University Open Access Fund
Keywords
Glucuronoyl esterase, Esterase, Cellobiosidase, Cellulose, Lignocellulose, Pichia pastoris
Citation
Conacher, C. G., et al. 2019. Scalable methanol‑free production of recombinant glucuronoyl esterase in Pichia pastoris. BMC Research Notes, 12: 596, doi:10.1186/s13104-019-4638-9