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Ziesack, M.

Publications and source records attributed to Ziesack, M..

4 recordsLinked to original sources
bioRxiv2021

Biocatalytic Formation of Novel Polyesters with para-Hydroxyphenyl groups in the Backbone - Engineering Cupriavidus necator for production of high-performance materials from CO2 and electricity

Synthetic materials are integral components of consumable and durable goods and are indispensable in the modern world. Polyesters are the most versatile bulk- and specialty-polymers but their production is not sustainable and their fate at end-of-life is of great concern. Bioplastics, though highly regarded, often lag behind conventional plastics due to poor material properties and a competitive market. This has limited the success of sustainable replacements at scale. Enabling the production of bioplastics with superior properties from waste-derived feedstocks could change that. To this end, we created a synthetic entry into the metabolic pathway of bio-polyester synthesis of Cupriavidus necator H16 employing heterologous hydroxyacyl-CoA transferase and mutant PHA synthase. The resulting microbial cell factories enabled the co-polymerization of a range of aliphatic and aromatic hydroxy carboxylates, including a hydroxyphenylic and a hydroxyfuranoic acid, for the first time incorporating aromatic rings in the backbone of biological polyesters. The latter were structurally analogous to synthetic polyesters like PET and PEF, as well as PBST and PBAT. The obtained bio-polyesters underwent characterization for specific physicochemical properties, complemented by the prediction of uncharacterized physicochemical and mechanical properties. In a further advance, the transgenic strain was cultivated in a bio-electrochemical system under autotrophic conditions, enabling synthesis of aromatic bio-polyesters with in situ generated H2+O2, while assimilating CO2. Employing elementary flux-mode analysis, metabolic modeling confirmed the feasibility of producing an extended range of aliphatic, arylatic, and aromatic PHAs de novo from various feedstocks. This comprehensive study paves the way toward sustainable bio-production of advanced high-performance thermoplastics and thermosets. Significance StatementBiomaterials can help the chemical industry transition to a carbon-neutral and circular economy, reducing the accumulation of greenhouse gases and plastic waste by developing biological replacements for fossil carbon-based plastics and implementing end-of-life strategies. Accomplished via the genetic engineering of a microbe that can assimilate carbon dioxide, this work demonstrates the first biocatalytic polymerization and incorporation of aromatic building blocks into the backbone of polyhydroxyalkanoates (PHAs). Employing a bio-electrochemical system for cultivation of the microbes, oxyhydrogen is formed and consumed in situ, thus avoiding explosive gas mixtures. The obtained aromatic PHAs are structurally similar and functionally analogs to synthetic bulk- and high-performance polymers such as PET (polyethylene terephthalate) and PEF (polyethylene furanoate) as well as PBST (polybutylene succinate-co-terephthalate) and PBAT (polybutylene adipate-co-terephthalate). Graphical AbstractThe biosynthesized novel aromatic polyhydroxyalkanoates described in this study are structurally similar to incumbent synthetic polyesters such as poly(ethylene terephthalate) (PET, orange box) and poly(ethylene furan-dicarboxylate) (PEF, yellow box). While PET and PEF may be bio-based to varying degrees, they are always synthesized chemically. In contrast, not only the monomers of poly(hydroxybutyrate-co-phloretate) and poly(hydroxybutyrate-co-hydroxymethylfuranoate) (green box) are fully bio-derived, but also the entire polymers are synthesized biologically. This was demonstrated by employing the autotrophic bacterium Cupriavidus necator H16. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/472320v3_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@20714borg.highwire.dtl.DTLVardef@1ee1c06org.highwire.dtl.DTLVardef@171cee7org.highwire.dtl.DTLVardef@c6bc21_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIFirst-ever biocatalytic formation of polyesters with aromatic rings in the backbone C_LIO_LIMicrobial production of aromatic PHAs in a bio-electrochemical system on CO2 and H2+O2 C_LIO_LIExpression-level of PHA synthase and molecular weight of polyesters are inversely correlated C_LIO_LIIn silico design and pathway analysis of bio-polyesters from low-cost carbon-feedstocks C_LIO_LIPrediction of material properties for novel aromatic co-polyesters and structural analogs C_LI

synthetic biology↗
bioRxiv2020

Valorization of CO2 through lithoautotrophic production of sustainable chemicals in Cupriavidus necator

Coupling recent advancements in genetic engineering of diverse microbes and gas-driven fermentation provides a path towards sustainable commodity chemical production. Cupriavidus necator H16 is a suitable species for this task because it effectively utilizes H2 and CO2 and is genetically tractable. Here, we demonstrate the versatility of C. necator for chemical production by engineering it to produce three products from CO2 under lithotrophic conditions: sucrose, polyhydroxyalkanoates (PHAs), and lipochitooligosaccharides (LCOs). We engineered sucrose production in a co-culture system with heterotrophic growth 30 times that of WT C. necator. We engineered PHA production (20-60% DCW) and selectively altered product composition by combining different thioesterases and phaCs to produce copolymers directly from CO2. And, we engineered C. necator to convert CO2 into the LCO, a plant growth enhancer, with titers of [~]1.4 mg/L--equivalent to yields in its native source, Bradyrhizobium. We applied the LCOs to germinating seeds as well as corn plants and observed increases in a variety of growth parameters. Taken together, these results expand our understanding f how a gas-utilizing bacteria can promote sustainable production.

synthetic biology↗
bioRxiv2018

Inducible Cooperation in a Synthetic Gut Bacterial Consortium Introduces Population Balance and Stability

In nature, microbes interact antagonistically, neutrally or beneficially. To shed light on the effects of positive interactions in microbial consortia we introduced metabolic dependencies and metabolite overproduction into four bacterial species. While antagonistic interactions govern the wildtype consortium behavior, the genetic modifications alleviated antagonistic interactions and resulted in beneficial interactions. Engineered cross-feeding increased population evenness, a component of ecological diversity, in different environments including in a more complex gnotobiotic mouse gut environment. Our findings suggest that metabolite cross-feeding could be used as a tool for intentionally shaping microbial consortia in complex environments.\n\nImportanceMicrobial communities are ubiquitous in nature. Bacterial consortia live in and on our body and in our environment and more recently, biotechnology is applying microbial consortia for bioproduction. As part of our body, bacterial consortia influence us in health and disease. Microbial consortia function is determined by its composition, which in turn is driven by the interactions between species. Further understanding of microbial interactions will help us deciphering how consortia function in complex environments and may enable us to modify microbial consortia for health and environmental benefits.

synthetic biology↗
bioRxiv2018

A synthetic bacterial information transfer system functions in the mammalian gut

The gut microbiome is intricately involved with establishing and maintaining the health of the host. Engineering of gut microbes aims to add new functions and expand the scope of control over the gut microbiome. To create systems that can perform increasingly complex tasks in the gut with multiple engineered strains it is necessary to program communication among these bacteria in the gut. Towards this goal, we engineered an information transfer system for inter-cellular communication, using native gut Escherichia coli and attenuated Salmonella enterica serovar Typhimurium. Specifically, we have taken two genetic circuits-one for signaling from the quorum sensing system and the other for memory from the bacteriophage genetic switch-and integrated them into a robust system that can report on successful communication in the mammalian gut. Our system provides a basis for the construction of a programmable gut consortia as well as a basis for further understanding of bacterial interactions in an otherwise hard-to-study environment.

synthetic biology↗