Dataset Information

Co-metabolism of thiocyanate and free cyanide by Exiguobacterium acetylicum and Bacillus marisflavi under alkaline conditions.

ABSTRACT: The continuous discharge of cyanide-containing effluents to the environment has necessitated for the development of environmentally benign treatment processes that would result in complete detoxification of the cyanide-containing wastewaters, without producing additional environmental toxicants. Since biological detoxification of hazardous chemical compounds has been renowned for its robustness and environmental-friendliness, the ability of the Exiguobacterium acetylicum (GenBank accession number KT282229) and Bacillus marisflavi (GenBank accession number KR016603) to co-metabolise thiocyanate (SCN-) and free cyanide (CN-) under alkaline conditions was evaluated. E. acetylicum had an SCN- degradation efficiency of 99.9 % from an initial SCN- concentration of 150 mg SCN-/L, but the organism was unable to degrade CN-. Consequently, B. marisflavi had a CN- degradation efficiency of 99 % from an initial concentration of 200 mg CN-/L. Similarly, the organism was unable to degrade SCN-; hence, this resulted in the evaluation of co-metabolism of SCN- and CN- by the two microbial species. Optimisation of operational conditions was evaluated using response surface methodology (RSM). A numeric optimisation technique was used to evaluate the optimisation of the input variables i.e. pH, temperature, SCN- and CN- concentrations. The optimum conditions were found to be as follows: pH 9.0, temperature 34 °C, 140 mg SCN-/L and 205 mg CN-/L under which complete SCN- and CN- degradation would be achieved over a 168-h period. Using the optimised data, co-metabolism of SCN- and CN- by both E. acetylicum and B. marisflavi was evaluated, achieving a combined degradation efficiency of ?99.9 %. The high degradative capacity of these organisms has resulted in their supplementation on an active continuous biological degradation system that is treating both SCN- and CN-.

SUBMITTER: Mekuto L

PROVIDER: S-EPMC4990519 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Co-metabolism of thiocyanate and free cyanide by Exiguobacterium acetylicum and Bacillus marisflavi under alkaline conditions.

Mekuto Lukhanyo L Alegbeleye Oluwadara Oluwaseun OO Ntwampe Seteno Karabo Obed SK Ngongang Maxwell Mewa MM Mudumbi John Baptist JB Akinpelu Enoch A EA

3 Biotech 20160818 2

The continuous discharge of cyanide-containing effluents to the environment has necessitated for the development of environmentally benign treatment processes that would result in complete detoxification of the cyanide-containing wastewaters, without producing additional environmental toxicants. Since biological detoxification of hazardous chemical compounds has been renowned for its robustness and environmental-friendliness, the ability of the Exiguobacterium acetylicum (GenBank accession numbe ...[more]

PMID: 28330245

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Project description:Three kinds of alkaliphilic bacteria able to utilize thiocyanate (CNS-) at pH 10 were found in highly alkaline soda lake sediments and soda soils. The first group included obligate heterotrophs that utilized thiocyanate as a nitrogen source while growing at pH 10 with acetate as carbon and energy sources. Most of the heterotrophic strains were able to oxidize sulfide and thiosulfate to tetrathionate. The second group included obligately autotrophic sulfur-oxidizing alkaliphiles which utilized thiocyanate nitrogen during growth with thiosulfate as the energy source. Genetic analysis demonstrated that both the heterotrophic and autotrophic alkaliphiles that utilized thiocyanate as a nitrogen source were related to the previously described sulfur-oxidizing alkaliphiles belonging to the gamma subdivision of the division Proteobacteria (the Halomonas group for the heterotrophs and the genus Thioalkalivibrio for autotrophs). The third group included obligately autotrophic sulfur-oxidizing alkaliphilic bacteria able to utilize thiocyanate as a sole source of energy. These bacteria could be enriched on mineral medium with thiocyanate at pH 10. Growth with thiocyanate was usually much slower than growth with thiosulfate, although the biomass yield on thiocyanate was higher. Of the four strains isolated, the three vibrio-shaped strains were genetically closely related to the previously described sulfur-oxidizing alkaliphiles belonging to the genus Thioalkalivibrio. The rod-shaped isolate differed from the other isolates by its ability to accumulate large amounts of elemental sulfur inside its cells and by its ability to oxidize carbon disulfide. Despite its low DNA homology with and substantial phenotypic differences from the vibrio-shaped strains, this isolate also belonged to the genus Thioalkalivibrio according to a phylogenetic analysis. The heterotrophic and autotrophic alkaliphiles that grew with thiocyanate as an N source possessed a relatively high level of cyanase activity which converted cyanate (CNO-) to ammonia and CO2. On the other hand, cyanase activity either was absent or was present at very low levels in the autotrophic strains grown on thiocyanate as the sole energy and N source. As a result, large amounts of cyanate were found to accumulate in the media during utilization of thiocyanate at pH 10 in batch and thiocyanate-limited continuous cultures. This is a first direct proof of a "cyanate pathway" in pure cultures of thiocyanate-degrading bacteria. Since it is relatively stable under alkaline conditions, cyanate is likely to play a role as an N buffer that keeps the alkaliphilic bacteria safe from inhibition by free ammonia, which otherwise would reach toxic levels during dissimilatory degradation of thiocyanate.

Project description:Significant progress has been made in isolating novel alkaline ?-mannanases, however, there is a paucity of information concerning the structural basis for alkaline tolerance displayed by these ?-mannanases. We report the catalytic domain structure of an industrially important ?-mannanase from the alkaliphilic Bacillus sp. N16-5 (BSP165 MAN) at a resolution of 1.6 Å. This enzyme, classified into subfamily 8 in glycosyl hydrolase family 5 (GH5), has a pH optimum of enzymatic activity at pH 9.5 and folds into a classic (?/?)(8)-barrel. In order to gain insight into molecular features for alkaline adaptation, we compared BSP165 MAN with previously reported GH5 ?-mannanases. It was revealed that BSP165 MAN and other subfamily 8 ?-mannanases have significantly increased hydrophobic and Arg residues content and decreased polar residues, comparing to ?-mannanases of subfamily 7 or 10 in GH5 which display optimum activities at lower pH. Further, extensive structural comparisons show alkaline ?-mannanases possess a set of distinctive features. Position and length of some helices, strands and loops of the TIM barrel structures are changed, which contributes, to a certain degree, to the distinctly different shaped (?/?)(8)-barrels, thus affecting the catalytic environment of these enzymes. The number of negatively charged residues is increased on the molecular surface, and fewer polar residues are exposed to the solvent. Two amino acid substitutions in the vicinity of the acid/base catalyst were proposed to be possibly responsible for the variation in pH optimum of these homologous enzymes in subfamily 8 of GH5, identified by sequence homology analysis and pK(a) calculations of the active site residues. Mutational analysis has proved that Gln91 and Glu226 are important for BSP165 MAN to function at high pH. These findings are proposed to be possible factors implicated in the alkaline adaptation of GH5 ?-mannanases and will help to further understanding of alkaline adaptation mechanism.

Dataset Information

Co-metabolism of thiocyanate and free cyanide by Exiguobacterium acetylicum and Bacillus marisflavi under alkaline conditions.

Publications

Co-metabolism of thiocyanate and free cyanide by Exiguobacterium acetylicum and Bacillus marisflavi under alkaline conditions.

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