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Title (Primary) Rational orthologous pathway and biochemical process engineering for adipic acid production using Pseudomonas taiwanensis VLB120
Author Bretschneider, L.; Heuschkel, I.; Bühler, K.; Karande, R.; Bühler, B.
Journal Metabolic Engineering
Year 2022
Department SOMA
Volume 70
Page From 206
Page To 217
Language englisch
Topic T7 Bioeconomy
Supplements https://ars.els-cdn.com/content/image/1-s2.0-S1096717622000209-mmc1.docx
Keywords Biotransformation; Enzyme cascade; Whole-cell biocatalyst; Nylon 6,6; Orthologous pathway engineering; Reaction engineering
Abstract Microbial bioprocessing based on orthologous pathways constitutes a promising approach to replace traditional greenhouse gas- and energy-intensive production processes, e.g., for adipic acid (AA). We report the construction of a Pseudomonas taiwanensis strain able to efficiently convert cyclohexane to AA. For this purpose, a recently developed 6-hydroxyhexanoic acid (6HA) synthesis pathway was amended with alcohol and aldehyde dehydrogenases, for which different expression systems were tested. Thereby, genes originating from Acidovorax sp. CHX100 and the XylS/Pm regulatory system proved most efficient for the conversion of 6HA to AA as well as the overall cascade enabling an AA formation activity of up to 48.6 ± 0.2 U gCDW−1. The optimization of biotransformation conditions enabled 96% conversion of 10 mM cyclohexane with 100% AA yield. During recombinant gene expression, the avoidance of glucose limitation was found to be crucial to enable stable AA formation. The biotransformation was then scaled from shaking flask to a 1 L bioreactor scale, at which a maximal activity of 22.6 ± 0.2 U gCDW−1 and an AA titer of 10.2 g L−1 were achieved. The principal feasibility of product isolation was shown by the purification of 3.4 g AA to a purity of 96.1%. This study presents the efficient bioconversion of cyclohexane to AA by means of a single strain and thereby sets the basis for an environmentally benign production of AA and related polymers such as nylon 6,6.
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=25696
Bretschneider, L., Heuschkel, I., Bühler, K., Karande, R., Bühler, B. (2022):
Rational orthologous pathway and biochemical process engineering for adipic acid production using Pseudomonas taiwanensis VLB120
Metab. Eng. 70 , 206 - 217