Abstract
Following biodegradation tests according to the OECD guidelines for testing of chemicals 301F different degradation rates were observed for the three stereoisomers of iminodisuccinate (IDS). A strain was isolated from activated sludge, which used two of three isomers, R,S-IDS and S,S-IDS, as sole source of carbon, nitrogen, and energy. The isolated strain was identified by 16S-rDNA and referred to as Ralstonia sp. SLRS7. An IDS-degrading lyase was isolated from the cell-free extract. The enzyme was purified by three chromatographic steps, which included anion-exchange chromatography, hydrophobic interaction chromatography and gel filtration. The lyase catalysed the non-hydrolytic cleavage of IDS without requirement of any cofactors. Cleavage of S,S-IDS led to the formation of fumaric acid and L-aspartic acid. Interestingly R,S-IDS yielded only D-aspartic acid besides fumaric acid. R,R-IDS was not transformed. Thus, the IDS-degrading enzyme is a carbon–nitrogen lyase attacking only the asymmetric carbon atom exhibiting the S-configuration. Besides S,S-IDS and R,S-IDS cleavage, the lyase catalysed also the transformation of certain S,S-IDS metal complexes, namely Ca2+-, Mg2+- and Mn2+-IDS. The maximum enzyme activity was found at pH 8.0–8.5 and 35 °C. SDS-PAGE analysis revealed a single 57-kDa protein band. The native enzyme was estimated to be around 240 kDa indicating a homotetramer enzyme.
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Cokesa, Ž., Lakner, S., Knackmuss, HJ. et al. A Stereoselective Carbon-Nitrogen Lyase from Ralstonia sp. SLRS7 Cleaves Two of Three Isomers of Iminodisuccinate. Biodegradation 15, 229–239 (2004). https://doi.org/10.1023/B:BIOD.0000042903.04718.f6
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DOI: https://doi.org/10.1023/B:BIOD.0000042903.04718.f6