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Co-expression of thermophilic pectinases in a single host for cost-effective pectin bioconversion into D-galacturonic acid

Flores-Fernández, Carol N., Cárdenas-Fernández, Max, Lye, Gary J., Ward, John M. (2023) Co-expression of thermophilic pectinases in a single host for cost-effective pectin bioconversion into D-galacturonic acid. Frontiers in Catalysis, 3 . Article Number 1112154. ISSN 2673-7841. (doi:10.3389/fctls.2023.1112154) (KAR id:100289)

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Co-expression of enzymes allow to produce multiple enzymes in a single host, representing a cost-effective alternative in biocatalytic processes which can be used for pectin bioconversion. Pectin-rich biomass is an abundant by-product from the fruit and sugar industries that is usually disposed in landfill or sold as a low value feedstock. The aim of this work was to co-express a thermophilic pectin methyl esterase (PME) and exo-polygalacturonases (exo-PGs) in a single host for pectin bioconversion into D-galacturonic acid (GalA) using different pectic substrates such as apple, citrus and sugar beet pectin. To achieve this, a PME from Bacillus licheniformis (BLI09) with either an exo-PG from Thermotoga maritima (TMA01) or from Bacillus licheniformis (BLI04) were cloned in pETDuet-1 and co-expressed in E. coli BL21 (DE3). Four co-expression plasmids containing both pectinases were constructed and factors such as the effect of the genes’ cloning order and their expression were evaluated. Co-expression constructs 3 and 4 (pETDuet-TMA01-BLI09 and pETDuet-BLI04-BLI09, respectively) showed better expression of both pectinases compared to co-expression constructs 1 and 2 (pETDuet-BLI09-TMA01 and pETDuet-BLI09-BLI04, respectively). Co-expression constructs 3 and 4 were the most efficient for pectin bioconversion into GalA reaching 3 and 2.5 mM GalA, respectively from apple and citrus pectin after 4 h reaction. In conclusion, this work demonstrates that the co-expression of pectinases can potentially contribute to reduce the cost associated to their production and purification as well as to increase their applicability for exploiting pectin-rich biomass to obtain bio-based chemicals.

Item Type: Article
DOI/Identification number: 10.3389/fctls.2023.1112154
Additional information: For the purpose of open access, the author(s) has applied a Creative Commons Attribution (CC BY) licence to any Author Accepted Manuscript version arising.
Uncontrolled keywords: Catalysis, enzymes co-expression, thermophilic pectinases, pectin, D-galacturonic acid, pectin methyl esterases, exo-polygalacturonases
Subjects: Q Science
Q Science > QH Natural history > QH301 Biology
Divisions: Divisions > Division of Natural Sciences > Biosciences
Funders: Biotechnology and Biological Sciences Research Council (
SWORD Depositor: JISC Publications Router
Depositing User: JISC Publications Router
Date Deposited: 03 Mar 2023 14:35 UTC
Last Modified: 20 Nov 2023 15:39 UTC
Resource URI: (The current URI for this page, for reference purposes)
Cárdenas-Fernández, Max:
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