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This Country Report on South Africa has been realised in the framework of the H2020-MSCA-RISE-2018 project “LoGov - Local Government and the Changing Urban-Rural Interplay”. LoGov aims to provide solutions for local governments that address the fundamental challenges resulting from urbanisation. To address this complex issue, 18 partners from 17 countries and six continents share their expertise and knowledge in the realms of public law, political science, and public administration. LoGov identifies, evaluates, compares, and shares innovative practices that cope with the impact of changing urban-rural relations in five major local government areas: (1) local responsibilities and public services, (2) local financial arrangements, (3) structure of local government, (4) intergovernmental relations of local governments, and (5) people’s participation in local decision-making. The present report provides an overview of the system of local government in South Africa. To access the full version of this report , the various practices in the five above-mentioned areas of interest, and to receive more information about the project, please visit: https://www.logov-rise.eu/. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 823961.

This entry has been realised in the framework of the H2020-MSCA-RISE-2018 project “LoGov - Local Government and the Changing Urban-Rural Interplay”. LoGov aims to provide solutions for local governments that address the fundamental challenges resulting from urbanisation. To address this complex issue, 18 partners from 17 countries and six continents share their expertise and knowledge in the realms of public law, political science, and public administration. LoGov identifies, evaluates, compares, and shares innovative practices that cope with the impact of changing urban-rural relations in five major local government areas: (1) local responsibilities and public services, (2) local financial arrangements, (3) structure of local government, (4) intergovernmental relations of local governments, and (5) people’s participation in local decision-making. The present entry addresses local responsibilities and public services in South Africa. The entry forms part of the LoGov Report on South Africa. To access the full version of the report on South Africa, other practices regarding responsibilities and public services and to receive more information about the project, please visit: https://www.logov-rise.eu/. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 823961.

Salicylic acid plays an important role in the plant immune response, and its degradation is therefore important for plant-pathogenic fungi. However, many nonpathogenic microorganisms can also degrade salicylic acid. In the filamentous fungus Aspergillus niger, two salicylic acid metabolic pathways have been suggested. The first pathway converts salicylic acid to catechol by a salicylate hydroxylase (ShyA). In the second pathway, salicylic acid is 3-hydroxylated to 2,3-dihydroxybenzoic acid, followed by decarboxylation to catechol by 2,3-dihydroxybenzoate decarboxylase (DhbA). A. niger cleaves the aromatic ring of catechol catalyzed by catechol 1,2-dioxygenase (CrcA) to form cis,cis-muconic acid. However, the identification and role of the genes and characterization of the enzymes involved in these pathways are lacking. In this study, we used transcriptome data of A. niger grown on salicylic acid to identify genes (shyA and crcA) involved in salicylic acid metabolism. Heterologous production in Escherichia coli followed by biochemical characterization confirmed the function of ShyA and CrcA. The combination of ShyA and CrcA demonstrated that cis,cis-muconic acid can be produced from salicylic acid. In addition, the in vivo roles of shyA, dhbA, and crcA were studied by creating A. niger deletion mutants which revealed the role of these genes in the fungal metabolism of salicylic acid. IMPORTANCE Nonrenewable petroleum sources are being depleted, and therefore, alternative sources are needed. Plant biomass is one of the most abundant renewable sources on Earth and is efficiently degraded by fungi. In order to utilize plant biomass efficiently, knowledge about the fungal metabolic pathways and the genes and enzymes involved is essential to create efficient strategies for producing valuable compounds such as cis,cis-muconic acid. cis,cis-Muconic acid is an important platform chemical that is used to synthesize nylon, polyethylene terephthalate (PET), polyurethane, resins, and lubricants. Currently, cis,cis-muconic acid is mainly produced through chemical synthesis from petroleum-based chemicals. Here, we show that two enzymes from fungi can be used to produce cis,cis-muconic acid from salicylic acid and contributes in creating alternative methods for the production of platform chemicals.

Additional file 1: Fig S1. Visualization of VdhA, VhyA and MhdA by SDS-PAGE. Fig. S2. Conversion of aromatic compounds by the reference (a), ∆vdhA (b), ∆vhyA (c), and ∆mhdA (d) after 24 h of incubation. Concentrations of the detected compounds can be found in Table 1. Error bars represent the standard deviation between three biological replicates. Fig. S3. Maximum likelihood (ML; 500 bootstraps) phylogenetic tree of A. niger VdhA compared to selected fungal genomes. The scale bar shows a distance equivalent to 0.2 amino acid substitutions per site. Values over 50% bootstrap support are shown with ML values in black, Neighbor Joining values in purple and Minimum Evolution values in blue. Characterized enzymes are in bold. Blue font represents ascomycete fungi, red font basidiomycete fungi, green font bacteria, and orange font Saccharomycetes. Fungal species names are followed by protein IDs from JGI ( http://genome.jgi-psf.org/programs/fungi/index.jsf ). Fig. S4. Maximum likelihood (ML; 500 bootstraps) phylogenetic tree of A. niger MhdA compared to selected fungal genomes. The scale bar shows a distance equivalent to 0.2 amino acid substitutions per site. Values over 50% bootstrap support are shown with ML values in black, Neighbor Joining values in purple and Minimum Evolution values in blue. In bold are characterized enzymes. Blue font represents ascomycete fungi, red font basidiomycete fungi, green font bacteria, black font plants and pink font Homo sapiens. Fungal species names are followed by protein IDs from JGI ( http://genome.jgi-psf.org/programs/fungi/index.jsf ). Fig. S5. 1H-NMR spectrum of 4-oxo-monomethyl adipate (D2O). The structure corresponds to 4-oxo-monomethyl adipate with D2 at C5 position.