Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium
Abstract This study reports the isolation and characterization of Bacillus subtilis K35-1, a novel cellulolytic strain with exceptional forage degradation capabilities. From eight B. subtilis isolates obtained from yak rumen fluid through Congo red screening (hydrolysis capacity = 2.61 ± 0.23), K35-...
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2025-07-01
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| Online Access: | https://doi.org/10.1186/s12866-025-04136-8 |
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| author | Dan Wu Jing Feng Dongxu Wen Hongzhuang Wang Sijia Lu Kun Li Rizwana Sultan Bin Li |
| author_facet | Dan Wu Jing Feng Dongxu Wen Hongzhuang Wang Sijia Lu Kun Li Rizwana Sultan Bin Li |
| author_sort | Dan Wu |
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| description | Abstract This study reports the isolation and characterization of Bacillus subtilis K35-1, a novel cellulolytic strain with exceptional forage degradation capabilities. From eight B. subtilis isolates obtained from yak rumen fluid through Congo red screening (hydrolysis capacity = 2.61 ± 0.23), K35-1 demonstrated superior enzymatic performance, achieving peak cellulase (77.26 U/mL) and hemicellulase (222.85 nmol/min/mL) activities at 36 h of fermentation. The whole genome sequencing revealed a 4.06 Mb circular chromosome (GC content 43.83%) encoding 3,980 protein-coding sequences. Comprehensive CAZy annotation identified 703 carbohydrate-active enzymes, including: 87 cellulases spanning 7 GH families (GH5, GH6, GH9, GH12, GH44, GH45, GH48) and 34 hemicellulases from 4 GH families (GH10, GH11, GH26, GH30). Comparative genomic analysis showed K35-1 possesses 40% more glycoside hydrolases than reference strains (Srivastava et al., Mol Genet Genomics 298:361–74, 2023), explaining its enhanced degradation efficiency (53.2% cellulose reduction vs. 7.3% in conventional treatments). Functional annotation revealed: 275 carbohydrate metabolism genes (KEGG), 228 cell wall/membrane biogenesis genes (COG) and 53.91% reduced-virulence mutations (PHI database). The strain’s robust enzymatic profile, coupled with minimal antibiotic resistance (11 genes, including ermB), positions K35-1 as both an efficient forage degrader and safe probiotic candidate. These findings provide a genomic foundation for developing novel feed additives to improve livestock nutrition. |
| format | Article |
| id | doaj-art-8bb311fe56ad40c1864478794261a1bb |
| institution | Kabale University |
| issn | 1471-2180 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj-art-8bb311fe56ad40c1864478794261a1bb2025-08-20T04:01:53ZengBMCBMC Microbiology1471-21802025-07-0125111010.1186/s12866-025-04136-8Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacteriumDan Wu0Jing Feng1Dongxu Wen2Hongzhuang Wang3Sijia Lu4Kun Li5Rizwana Sultan6Bin Li7State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, and Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry SciencesState Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, and Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry SciencesState Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, and Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry SciencesState Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, and Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry SciencesCollege of Veterinary Medicine, Nanjing Agricultural UniversityCollege of Veterinary Medicine, Nanjing Agricultural UniversityDepartment of Pathology, Faculty of Veterinary Science, Cholistan University of Veterinary and Animal Sciences (CUVAS)State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, and Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry SciencesAbstract This study reports the isolation and characterization of Bacillus subtilis K35-1, a novel cellulolytic strain with exceptional forage degradation capabilities. From eight B. subtilis isolates obtained from yak rumen fluid through Congo red screening (hydrolysis capacity = 2.61 ± 0.23), K35-1 demonstrated superior enzymatic performance, achieving peak cellulase (77.26 U/mL) and hemicellulase (222.85 nmol/min/mL) activities at 36 h of fermentation. The whole genome sequencing revealed a 4.06 Mb circular chromosome (GC content 43.83%) encoding 3,980 protein-coding sequences. Comprehensive CAZy annotation identified 703 carbohydrate-active enzymes, including: 87 cellulases spanning 7 GH families (GH5, GH6, GH9, GH12, GH44, GH45, GH48) and 34 hemicellulases from 4 GH families (GH10, GH11, GH26, GH30). Comparative genomic analysis showed K35-1 possesses 40% more glycoside hydrolases than reference strains (Srivastava et al., Mol Genet Genomics 298:361–74, 2023), explaining its enhanced degradation efficiency (53.2% cellulose reduction vs. 7.3% in conventional treatments). Functional annotation revealed: 275 carbohydrate metabolism genes (KEGG), 228 cell wall/membrane biogenesis genes (COG) and 53.91% reduced-virulence mutations (PHI database). The strain’s robust enzymatic profile, coupled with minimal antibiotic resistance (11 genes, including ermB), positions K35-1 as both an efficient forage degrader and safe probiotic candidate. These findings provide a genomic foundation for developing novel feed additives to improve livestock nutrition.https://doi.org/10.1186/s12866-025-04136-8Bacillus subtilisCellulose degrading bacteriaWhole genome |
| spellingShingle | Dan Wu Jing Feng Dongxu Wen Hongzhuang Wang Sijia Lu Kun Li Rizwana Sultan Bin Li Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium BMC Microbiology Bacillus subtilis Cellulose degrading bacteria Whole genome |
| title | Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium |
| title_full | Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium |
| title_fullStr | Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium |
| title_full_unstemmed | Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium |
| title_short | Identification and whole genome sequencing analysis of Bacillus subtilis K35-1, a highly efficient cellulose in forage degrading bacterium |
| title_sort | identification and whole genome sequencing analysis of bacillus subtilis k35 1 a highly efficient cellulose in forage degrading bacterium |
| topic | Bacillus subtilis Cellulose degrading bacteria Whole genome |
| url | https://doi.org/10.1186/s12866-025-04136-8 |
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