Abstract
Keratin-rich wastes, mainly in the form of feathers, are recalcitrant residues generated in high amounts as by-products in chicken farms and food industry. Polylactic acid (PLA) is the second most common biodegradable polymer found in commercial plastics, which is not easily degraded by microbial activity. This work reports the 3.8-Mb genome of Bacillus altitudinis B12, a highly efficient PLA- and keratin-degrading bacterium, with potential for environmental friendly biotechnological applications in the feed, fertilizer, detergent, leather, and pharmaceutical industries. The whole genome sequence of B. altitudinis B12 revealed that this strain (which had been previously misclassified as Bacillus pumilus B12) is closely related to the B. altitudinis strains ER5, W3, and GR-8. A total of 4056 coding sequences were annotated using the RAST server, of which 2484 are core genes of the pan genome of B. altitudinis and 171 are unique to this strain. According to the sequence analysis, B. pumilus B12 has a predicted secretome of 353 proteins, among which a keratinase and a PLA depolymerase were identified by sequence analysis. The presence of these two enzymes could explain the characterized PLA and keratin biodegradation capability of the strain.
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Data availability
The assembled genome has been deposited at GenBank under the accession number CP101506 (BioProject: PRJNA858691). All the other relevant files for the genome and pan genome analyses are available at: https://github.com/SergioBordel/Bacillus_altitudinis_B12.
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Acknowledgements
We thank Prof. Todd B. Reynolds (University of Tennessee, USA) for providing the strain.
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The FEDER program (TCUE 2021–2023) is gratefully acknowledged for funding the project with reference number: 067/229111 to Dr. Fernando Santos-Beneit.
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438_2022_1989_MOESM1_ESM.pptx
Supplementary file1 Supplementary Fig. S1. Bacillus altitudinis B12 and Bacillus subtilis 168 PLA degrading activity on different media compositions. Cultures of 100 mL were grown in 1 L glass bottles using MSM medium (with either 1% glucose or mannitol as sole carbon source) at 30 ℃ and 180 rpm for 72 h. All cultures contained 2 % of extruded granules of PLA (~ 500 µm). After 72 h, lactate release and OD600 was measured by HPLC and spectrophotometry, respectively. Note the differences in lactate release when growing B. altitudinis B12 in glucose or mannitol (PPTX 60 KB)
438_2022_1989_MOESM2_ESM.pptx
Supplementary file2 Supplementary Fig. S2. A Histograms showing the numbers of genes versus the number of strains in which they have orthologues. The different panels show histograms for the pan genome, and for the genes in the strains B12, ER5 and H5-9. B Histogram showing how the number of unique genes is distributed among different strains (PPTX 148 KB)
438_2022_1989_MOESM3_ESM.pptx
Supplementary file3 Supplementary Fig. S3. Secondary metabolite biosynthetic clusters identified by antiSMASH version 6.0. Note the three cluster showing > 50 % similarity to the known biosynthetic gene cluster of the lipopeptides lichenysin, fengycin and bacilysin (PPTX 230 KB)
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Bordel, S., Martín-González, D., Muñoz, R. et al. Genome sequence analysis and characterization of Bacillus altitudinis B12, a polylactic acid- and keratin-degrading bacterium. Mol Genet Genomics 298, 389–398 (2023). https://doi.org/10.1007/s00438-022-01989-w
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DOI: https://doi.org/10.1007/s00438-022-01989-w