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Title: | Cloning and expression of β-1, 4-endoglucanase gene from Bacillus subtilis isolated from soil long term irrigated with effluents of paper and pulp mill |
Other Titles: | Not Available |
Authors: | Pandey, S., Kushwah J., Tiwari, R. Kumar, R., Somvanshi, V.S., Nain, L Saxena, A.K |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR-NATIONAL BUREAU OF AGRICULTURALLY IMPORTANT MICROORGANISMS KUSHMAUR MAU NATH BHANJAN 275103 |
Published/ Complete Date: | 2014-01-01 |
Project Code: | Not Available |
Keywords: | Cellulaseβ-1, 4-Endoglucanase; Cloning;Overexpression |
Publisher: | ELSEVIER |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | A strain of Bacillus subtilis IARI-SP-1 isolated from soil long term irrigated with effluents of paper and pulp mill showed high β-1, 4-endoglucanase (2.5 IU/ml) but low activity of β-1, 4-exoglucanase (0.8 IU/ml) and β-glucosidase (0.084 IU/ml). The β-1, 4-endoglucanase gene of IARI-SP-1 was amplified using degenerate primers designed based on sequences already available in NCBI GenBank. A full length gene of β-1, 4-endonuclease consisting of 1499 nucleotides was identified through sequence analysis of the amplified product. The ORF encoded for a protein of 500 amino acids with a predicted molecular weight of 55 kDa. The gene was cloned in pET-28a and over expressed in Escherichia coli BL21 (DE3). In comparison to wild strain (B. subtilis), the transformed E. coli exhibited four times increase in cellulase production. Higher enzyme activity was observed in supernatant (8.2 IU/ml) than cell pellet (2.8 IU/ml) suggesting more extracellular production of β-1, 4-endoglucanase. SDS-PAGE and CMC plate assay also confirmed the overproduction by the transformed E. coli. The pH and temperature optima of expressed β-1, 4-endoglucanase enzyme was identical to that of wild strain and was 8 and 50–60 °C, respectively. |
Description: | Not Available |
ISSN: | Not Available |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | Microbiological Research (Zentralblatt f?r Mikrobiologie) |
NAAS Rating: | 9.97 |
Volume No.: | 169 |
Page Number: | 693-698 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | Not Available |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/65734 |
Appears in Collections: | CS-NBAIMO-Publication |
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