Die fermentative Herstellung von Biotensiden wird meist durch die exzessive Bildung von stabilem Schaum erschwert. Am Beispiel von Monorhamnolipiden werden Vorschläge sowohl zur stabilen Aufrechterhaltung von begasten und gerührten Fermentationen wie auch zur Isolierung dieser Biotenside im größeren Maßstab gegeben, da diese kommerziell nur in sehr kleinen Mengen erhältlich sind. Ein besonderer Fokus liegt dabei auf der Verwendung nachwachsender Rohstoffe.

Fusicocca-2,10(14)-diene (FCdiene) is a diterpene which is interesting as a precursor of the anti-cancer drug fusicoccin A and therefore for pharmaceutical applications. Production of FCdiene using a genetically modified Saccharomyces cerevisiae has been previously demonstrated with batch cultivations with a product concentration up to 10 mg/L. However, it is widely known that fed-batch processes can significantly improve product titer in yeast fermentations. This study focuses on the establishment of fed-batch fermentation for FCdiene production because fed-batch cultivations using FeedBeads® indicated that limiting glucose supply could increase yields of biomass (1.07 gCDW/gGlucose instead of 0.20 gCDW/gGlucose) and FCdiene (21.54 mgFCdiene/gGlucose instead of 9.74 mgFCdiene/gGlucose) in shake flask scale and may have implications for larger scale processes. We implemented a new exponential glucose feed profile in a 1.8 L stirred tank reactor. This reduced overfeeding and the consequent, ethanol production. As a result improvements in cell concentrations up to 246% could be achieved and FCdiene yield increased up to 2.8X in the first 28 h. FCdiene concentration reached 161 mg/L and 320 mg/L at 44 h. Fed-batch and batch mode were combined to examine dynamics of bi-modal cultivation where a fed-batch phase was used for biomass production and a batch phase used for FCdiene production potentially supported by ethanol consumption as reported on production of betulinic acid. The present study highlights the potential of process development improvements which increase high-value heterologous diterpene yields from S. cerevisiae.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0662-8) contains supplementary material, which is available to authorized users.

Towards establishing a prospective industrial microbial lignan production process, we set up and investigated the biotransformation of coniferyl alcohol to secoisolariciresinol with recombinant Escherichia coli in a stirred-tank reactor (STR). Initially, we tested different cofactor concentrations and antifoam additions in shake flasks. Next, we designed an STR batch bioprocess and tested aeration rates, pH regulation, and substrate-feeding strategies. Targeted metabolomics of phenylpropanoids and lignans assisted the bioprocess development by monitoring the lignan pathway activity. We found that the copper concentration and the substrate-feeding strategy had considerable impact on lignan production. Furthermore, time-resolved monitoring of pathway metabolites revealed two maximal intracellular lignan concentrations, the first shortly after induction of gene expression and the second after the cells entered the stationary growth phase. During STR cultivation, a maximal intracellular titer of 130.4 mg L−1 secoisolariciresinol was achieved, corresponding to a yield coefficient of 26.4 mg g−1 and a space–time yield of 2.6 mg L−1 h−1. We report for the first time the in-depth evaluation of microbially produced lignans in a well-controlled STR bioprocess. Monitoring of the lignan pathway activity showed that lignan accumulation is highly dynamic during the cultivation and points towards the need for a more efficient coniferyl alcohol dimerization system for optimal microbial production conditions.

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