Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis
Abstract Background Saccharification is the most crucial and cost-intensive process in second generation biofuel production. The deficiency of β-glucosidase in commercial enzyme leads to incomplete biomass hydrolysis. The decomposition of biomass at high temperature environments leads us to isolate...
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| Format: | Article |
| Language: | English |
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BMC
2017-10-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-017-0932-8 |
| _version_ | 1811244934721699840 |
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| author | Rameshwar Tiwari Puneet Kumar Singh Surender Singh Pawan K. S. Nain Lata Nain Pratyoosh Shukla |
| author_facet | Rameshwar Tiwari Puneet Kumar Singh Surender Singh Pawan K. S. Nain Lata Nain Pratyoosh Shukla |
| author_sort | Rameshwar Tiwari |
| collection | DOAJ |
| description | Abstract Background Saccharification is the most crucial and cost-intensive process in second generation biofuel production. The deficiency of β-glucosidase in commercial enzyme leads to incomplete biomass hydrolysis. The decomposition of biomass at high temperature environments leads us to isolate thermotolerant microbes with β-glucosidase production potential. Results A total of 11 isolates were obtained from compost and cow dung samples that were able to grow at 50 °C. On the basis of qualitative and quantitative estimation of β-glucosidase enzyme production, Bacillus subtilis RA10 was selected for further studies. The medium components and growth conditions were optimized and β-glucosidase enzyme production was enhanced up to 19.8-fold. The β-glucosidase from B. subtilis RA10 retained 78% of activity at 80 °C temperature and 68.32% of enzyme activity was stable even at 50 °C after 48 h of incubation. The supplementation of β-glucosidase from B. subtilis RA10 into commercial cellulase enzyme resulted in 1.34-fold higher glucose release. Furthermore, β-glucosidase was also functionally elucidated by cloning and overexpression of full length GH1 family β-glucosidase gene from B. subtilis RA10. The purified protein was characterized as thermostable β-glucosidase enzyme. Conclusions The thermostable β-glucosidase enzyme from B. subtilis RA10 would facilitate efficient saccharification of cellulosic biomass into fermentable sugar. Consequently, after saccharification, thermostable β-glucosidase enzyme would be recovered and reused to reduce the cost of overall bioethanol production process. |
| first_indexed | 2024-04-12T14:34:05Z |
| format | Article |
| id | doaj.art-33d13d967bd84076933557486646e8dd |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-12T14:34:05Z |
| publishDate | 2017-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-33d13d967bd84076933557486646e8dd2022-12-22T03:29:10ZengBMCBiotechnology for Biofuels1754-68342017-10-0110111810.1186/s13068-017-0932-8Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysisRameshwar Tiwari0Puneet Kumar Singh1Surender Singh2Pawan K. S. Nain3Lata Nain4Pratyoosh Shukla5Enzyme Technology and Protein Bioinformatics Laboratory, Department of Microbiology, Maharshi Dayanand UniversityEnzyme Technology and Protein Bioinformatics Laboratory, Department of Microbiology, Maharshi Dayanand UniversityDivision of Microbiology, Indian Agricultural Research InstituteDesign and Mechatronic Division, School of Civil and Mechanical Engineering, Galgotias UniversityDivision of Microbiology, Indian Agricultural Research InstituteEnzyme Technology and Protein Bioinformatics Laboratory, Department of Microbiology, Maharshi Dayanand UniversityAbstract Background Saccharification is the most crucial and cost-intensive process in second generation biofuel production. The deficiency of β-glucosidase in commercial enzyme leads to incomplete biomass hydrolysis. The decomposition of biomass at high temperature environments leads us to isolate thermotolerant microbes with β-glucosidase production potential. Results A total of 11 isolates were obtained from compost and cow dung samples that were able to grow at 50 °C. On the basis of qualitative and quantitative estimation of β-glucosidase enzyme production, Bacillus subtilis RA10 was selected for further studies. The medium components and growth conditions were optimized and β-glucosidase enzyme production was enhanced up to 19.8-fold. The β-glucosidase from B. subtilis RA10 retained 78% of activity at 80 °C temperature and 68.32% of enzyme activity was stable even at 50 °C after 48 h of incubation. The supplementation of β-glucosidase from B. subtilis RA10 into commercial cellulase enzyme resulted in 1.34-fold higher glucose release. Furthermore, β-glucosidase was also functionally elucidated by cloning and overexpression of full length GH1 family β-glucosidase gene from B. subtilis RA10. The purified protein was characterized as thermostable β-glucosidase enzyme. Conclusions The thermostable β-glucosidase enzyme from B. subtilis RA10 would facilitate efficient saccharification of cellulosic biomass into fermentable sugar. Consequently, after saccharification, thermostable β-glucosidase enzyme would be recovered and reused to reduce the cost of overall bioethanol production process.http://link.springer.com/article/10.1186/s13068-017-0932-8ThermotolerantThermostable β-glucosidaseSaccharificationCloning and expressionMolecular modelling |
| spellingShingle | Rameshwar Tiwari Puneet Kumar Singh Surender Singh Pawan K. S. Nain Lata Nain Pratyoosh Shukla Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis Biotechnology for Biofuels Thermotolerant Thermostable β-glucosidase Saccharification Cloning and expression Molecular modelling |
| title | Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis |
| title_full | Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis |
| title_fullStr | Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis |
| title_full_unstemmed | Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis |
| title_short | Bioprospecting of novel thermostable β-glucosidase from Bacillus subtilis RA10 and its application in biomass hydrolysis |
| title_sort | bioprospecting of novel thermostable β glucosidase from bacillus subtilis ra10 and its application in biomass hydrolysis |
| topic | Thermotolerant Thermostable β-glucosidase Saccharification Cloning and expression Molecular modelling |
| url | http://link.springer.com/article/10.1186/s13068-017-0932-8 |
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