Combined Surface-Subsurface Stream Restoration Structures Can Optimize Hyporheic Attenuation of Stream Water Contaminants.

There is a design-to-function knowledge gap regarding how engineered stream restoration structures can maximize hyporheic contaminant attenuation. Surface and subsurface structures have each been studied in isolation as techniques to restore hyporheic exchange, but surface-subsurface structures have not been investigated or optimized in an integrated manner. Here, we used a numerical model to systematically evaluate key design variables for combined surface (i.e., weir height and length) and subsurface (i.e., upstream and downstream baffle plate spacing) structures. We also compared performance metrics that place differing emphasis on hyporheic flux versus transit times. We found that surface structures tended to create higher flux, shorter transit time flowpaths, whereas subsurface structures promoted moderate-flux, longer transit time flowpaths. Optimal combined surface-subsurface structures could increase fluxes and transit times simultaneously, thus providing conditions for contaminant attenuation that were many times more effective than surface or subsurface structures alone. All performance metrics were improved by the presence of an upstream plate and the absence of a downstream plate. Increasing the weir length tended to improve all metrics, whereas the optimal weir height varied based on metrics. These findings may improve stream restoration by better aligning specific restoration goals with appropriate performance metrics and hyporheic structure designs.

Vinylogous chain branching catalysed by a dedicated polyketide synthase module.

Bacteria use modular polyketide synthases (PKSs) to assemble complex polyketides, many of which are leads for the development of clinical drugs, in particular anti-infectives and anti-tumoral agents. Because these multifarious compounds are notoriously difficult to synthesize, they are usually produced by microbial fermentation. During the past two decades, an impressive body of knowledge on modular PKSs has been gathered that not only provides detailed insight into the biosynthetic pathways but also allows the rational engineering of enzymatic processing lines to yield structural analogues. Notably, a hallmark of all PKS modules studied so far is the head-to-tail fusion of acyl and malonyl building blocks, which leads to linear backbones. Yet, structural diversity is limited by this uniform assembly mode. Here we demonstrate a new type of PKS module from the endofungal bacterium Burkholderia rhizoxinica that catalyses a Michael-type acetyl addition to generate a branch in the carbon chain. In vitro reconstitution of the entire PKS module, X-ray structures of a ketosynthase-branching didomain and mutagenesis experiments revealed a crucial role of the ketosynthase domain in branching the carbon chain. We present a trapped intermediary state in which acyl carrier protein and ketosynthase are covalently linked by the branched polyketide and suggest a new mechanism for chain alkylation, which is functionally distinct from terpenoid-like ß-branching. For the rice seedling blight toxin rhizoxin, one of the strongest known anti-mitotic agents, the non-canonical polyketide modification is indispensable for phytotoxic and anti-tumoral activities. We propose that the formation of related pharmacophoric groups follows the same general scheme and infer a unifying vinylogous branching reaction for PKS modules with a ketosynthase-branching-acyl-carrier-protein architecture. This study unveils the structure and function of a new PKS module that broadens the biosynthetic scope of polyketide biosynthesis and sets the stage for rationally creating structural diversity.

Assessing oxazole bioisosteres as mutasynthons on the rhizoxin assembly line.

Chain armor against tumor cells: The oxazole side chain in the antimitotic agent rhizoxin S2 (1) was successfully replaced through mutasynthesis by using an engineered mutant impaired in heterocyclization. Incorporation of 12 non-natural surrogates into fully processed rhizoxin analogues revealed a remarkable substrate flexibility of the PKS-NRPS hybrid.

The auxin herbicide mecoprop-P in new light: Filling the data gap for dicotyledonous macrophytes.

Mecoprop-P (MCPP-P) is an auxin herbicide which has been used against dicotyledonous weed plants since the 1980s. While fate and monitoring data of MCPP-P in the aquatic environment revealing concentrations up to 103 µg/L in freshwaters are well documented, only very few toxicity data and no studies with dicotyledonous macrophytes have been published in open literature so far. To fill up this essential data gap, a microcosm study was conducted in order to test the sensitivity of nine dicotyledonous and one Ceratophyllales macrophyte species. The plant species were exposed to seven MCPP-P concentrations ranging from 8 to 512 µg/L for 21/22 days in one microcosm per concentration, and two further microcosms served as controls. Plant preparation was adapted to each species and endpoints were measured to calculate growth rates. Data were generated to obtain effect concentrations (ECX) which then were used to construct species sensitivity distribution curves (SSD). Eight species proved to be sensitive to MCPP-P in the tested concentration range with EC50 values ranging from 46.9 µg/L for Ranunculus aquatilis to 656.4 µg/L MCPP-P for Ludwigia repens. Taking the EC50 values of this study and published data for autotrophic organisms into account, a hazard concentration (HC5) of 2.7 µg/L was derived from the SSD curve, while an SSD curve without dicotyledonous macrophytes resulted in an about 100 times higher HC5 (360.8 µg/L MCCP-P). This confirms that a re-evaluation for old auxin herbicides by including dicotyledonous test species into the environmental risk assessment may be indicated. Furthermore, the use of MCPP-P in bitumen felts as protection against rooting by plants is not in the focus of any risk regulation so far. This application, however, can lead to high run-off concentrations that can enter surface waters easily, exceeding the new regulatory acceptable concentration values.

Polyketide-chain branching by an enzymatic Michael addition.

A new "branch" for polyketide synthases was discovered in the biosynthesis of the antimitotic rhizoxin complex in the endofungal bacterium Burkholderia rhizoxinica. Genetic engineering and the structural elucidation of pathway intermediates revealed that a complex polyketide chain is branched at the beta position by an unprecedented conjugate addition of an acetyl building block to an acryloyl precursor (see scheme).