EXPRESSION OF CELLULOSE-DEGRADING ENDOGLUCANASE FROM BACILLUS SUBTILIS USING PTOLT EXPRESSION SYSTEM IN ESCHERICHIA COLI
| Autor: | Hülya Kuduğ Ceylan, Yakup Ulusu, İsa Gökçe, Sema Bilgin |
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| Přispěvatelé: | Ulusu, Yakup |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Ptolt Escherichia Coli biology 010405 organic chemistry Organic Chemistry Bacillus subtilis Cellulase Recombinant Protein biology.organism_classification medicine.disease_cause 01 natural sciences 0104 chemical sciences chemistry.chemical_compound Biochemistry chemistry 010608 biotechnology Materials Chemistry biology.protein medicine Endoglucanase Cellulose Escherichia coli |
| Zdroj: | Cellulose Chemistry and Technology. 55:619-627 |
| ISSN: | 2457-9459 0576-9787 0006-7768 |
| Popis: | WOS:000677681400010 Endoglucanases randomly hydrolyse the cellulose chains by acting upon internal beta-1,4-D-glycosidic bonds and are used extensively in industrial applications. In this study, bacterial endoglucanase gene yhfE was obtained by PCR, using primers based on genomic sequences of Bacillus subtilis strains. 1041 bp DNA fragment of yhfE was cloned into Escherichia coli DH5 alpha through the use of pTolT expression plasmid. PCR, restriction enzyme analysis and DNA sequencing were performed in order to confirm the cloning. E. coli BL21-AI cells expressed the yhfE after induction at 0.04% of arabinose concentration for 4 h. The expected 38.7 kDa size yhfE protein after digestion with thrombin of the His-tagged fusion protein (yhfE-TolAIII) was visualized by SDS-PAGE. The yhfE-TolAIII production yield was approximately 82 mg/L. The recombinant yhfE was characterized by MALDI-TOF mass spectrometry and CD analysis. |
| Databáze: | OpenAIRE |
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