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Bioflocculation potentials of a uronic acid-containing glycoprotein produced by Bacillus sp. AEMREG4 isolated from Tyhume River, South Africa.

ABSTRACT: Bioflocculants are secondary metabolites produced by microorganisms during their growth which have received attentions due to their biodegradability, innocuousness and lack of secondary pollution from degradation intermediates. This study reports on a bioflocculant produced by Bacillus specie isolated from Thyume River in South Africa. The bacterial isolate was identified through 16S rDNA sequencing and the BLAST analysis of the nucleotide sequences revealed 99% similarity to Bacillus sp. BCT-7112. The sequence was subsequently deposited in the GenBank as Bacillus sp. AEMREG4 with accession number KP406729. The optimum culture conditions for bioflocculant production were an inoculum size 4% (v/v) (80%) and starch (81%) as well as yeast extract (82%) as sole carbon and nitrogen sources, respectively. Addition of Ca2+ greatly enhanced the flocculating activity (76%) of crude bioflocculant over a wide range of pH 4-10 and retained high flocculating activity when heated at 100 °C for 1 h. Chemical analyses of the purified bioflocculant revealed carbohydrate (79% w/w) as a predominant component followed by uronic acid (15% w/w) and protein (5% w/w). Fourier transform infrared spectrum revealed the presence of carboxyl, hydroxyl and methoxyl groups as the functional groups responsible for flocculation and the high flocculation activity achieved portends its industrial applicability.

SUBMITTER: Ntsangani N

PROVIDER: S-EPMC5429313 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Bioflocculation potentials of a uronic acid-containing glycoprotein produced by Bacillus sp. AEMREG4 isolated from Tyhume River, South Africa.

Ntsangani Nozipho N Okaiyeto Kunle K Uchechukwu Nwodo U NU Olaniran Ademola O AO Mabinya Leonard V LV Okoh Anthony I AI

3 Biotech 20170512 1

Bioflocculants are secondary metabolites produced by microorganisms during their growth which have received attentions due to their biodegradability, innocuousness and lack of secondary pollution from degradation intermediates. This study reports on a bioflocculant produced by Bacillus specie isolated from Thyume River in South Africa. The bacterial isolate was identified through 16S rDNA sequencing and the BLAST analysis of the nucleotide sequences revealed 99% similarity to Bacillus sp. BCT-71 ...[more]

PMID: 28500400

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Project description:BackgroundThe genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C30-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species' genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC-MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C30-carotenoid fatty acid ester produced by Planococcus sp. CP5-4.ResultsGenome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and "yellow" mutants respectively. Eight candidate genes, likely responsible for C30-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the "yellow" mutant phenotype. Genome annotation revealed that the genes encoded 4,4'-diapolycopene oxygenase (CrtNb), 4,4'- diapolycopen-4-al dehydrogenase (CrtNc), 4,4'-diapophytoene desaturase (CrtN), 4,4'- diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C30-carotenoid fatty acid ester, methyl 5-(6-C17:3)-glucosyl-5, 6'-dihydro-apo-4, 4'-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4.ConclusionWe identified and characterized the carotenoid biosynthetic gene cluster and the C30-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4'-lycopenoate esterified to heptadecatrienoic acid (C17:3). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show that various C30-carotenoid product chemotypes, apart from methyl 5-glucosyl-5, 6-dihydro-apo-4, 4'-lycopenoate and 5-glucosyl-4, 4-diaponeurosporen-4'-ol-4-oic acid, may be produced that could offer opportunities for a variety of applications.

Project description:BackgroundOligosaccharides from polysaccharides containing uronic acids are known to have many useful bioactivities. Thus, polysaccharide lyases (PLs) and glycoside hydrolases (GHs) involved in producing the oligosaccharides have attracted interest in both medical and industrial settings. The numerous polysaccharide lyases and glycoside hydrolases involved in producing the oligosaccharides were isolated from soil and marine microorganisms. Our previous report demonstrated that an agar-degrading bacterium, Catenovulum sp. CCB-QB4, isolated from a coastal area of Penang, Malaysia, possessed 183 glycoside hydrolases and 43 polysaccharide lyases in the genome. We expected that the strain might degrade and use uronic acid-containing polysaccharides as a carbon source, indicating that the strain has a potential for a source of novel genes for degrading the polysaccharides.MethodsTo confirm the expectation, the QB4 cells were cultured in artificial seawater media with uronic acid-containing polysaccharides, namely alginate, pectin (and saturated galacturonate), ulvan, and gellan gum, and the growth was observed. The genes involved in degradation and utilization of uronic acid-containing polysaccharides were explored in the QB4 genome using CAZy analysis and BlastP analysis.ResultsThe QB4 cells were capable of using these polysaccharides as a carbon source, and especially, the cells exhibited a robust growth in the presence of alginate. 28 PLs and 22 GHs related to the degradation of these polysaccharides were found in the QB4 genome based on the CAZy database. Eleven polysaccharide lyases and 16 glycoside hydrolases contained lipobox motif, indicating that these enzymes play an important role in degrading the polysaccharides. Fourteen of 28 polysaccharide lyases were classified into ulvan lyase, and the QB4 genome possessed the most abundant ulvan lyase genes in the CAZy database. Besides, genes involved in uronic acid metabolisms were also present in the genome. These results were consistent with the cell growth. In the pectin metabolic pathway, the strain had genes for three different pathways. However, the growth experiment using saturated galacturonate exhibited that the strain can only use the pathway related to unsaturated galacturonate.

Dataset Information

Bioflocculation potentials of a uronic acid-containing glycoprotein produced by Bacillus sp. AEMREG4 isolated from Tyhume River, South Africa.

Publications

Bioflocculation potentials of a uronic acid-containing glycoprotein produced by Bacillus sp. AEMREG4 isolated from Tyhume River, South Africa.

Similar Datasets

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