Electronic Resource
Structural elucidation and molecular characterization of Marinobacter sp. α-amylase
| العنوان: | Structural elucidation and molecular characterization of Marinobacter sp. α-amylase |
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| المؤلفون: | Sumit Kumar, Rizwan Hasan Khan, S. K. Khare |
| بيانات النشر: | Figshare 2016 Created: 2016-04-13 Updated: 2016-04-26 |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | Halophiles have been perceived as potential source of novel enzymes in recent years. The interest emanates from their ability to catalyze efficiently under high salt and organic solvents. Marinobacter sp. EMB8 α-amylase was found to be active and stable in salt and organic solvents. A study was carried out using circular dichroism (CD), fluorescence spectroscopy, and bioinformatics analysis of similar protein sequence to ascertain molecular basis of salt and solvent adaptability of α-amylase. Structural changes recorded in the presence of varying amounts of NaCl exhibited an increase in negative ellipticity as a function of salt, confirming that salt stabilizes the protein and increases the secondary structure, making it catalytically functional. The data of intrinsic and extrinsic fluorescence (using 1-anilinonaphthalene 8-sulfonate [ANS] as probe) further confirmed the role of salt. The α-amylase was active in the presence of nonpolar solvents, namely, hexane and decane, but inactivated by ethanol. The decrease in the activity was correlated with the loss of tertiary structure in the presence of ethanol. Guanidine hydrochloride and pH denaturation indicated the molten globule state at pH 4.0. Partial N-terminal amino acid sequence of the purified α-amylase revealed the relatedness to Pseudoalteromonas sp. α-amylase. “FVHLFEW” was found as the N-terminal signature sequence. Bioinformatics analysis was done using M. algicola α-amylase protein having the same N-terminal signature sequence. The three-dimensional structure of Marinobacter α-amylase was deduced using the I-TASSER server, which reflected the enrichment of acidic amino acids on the surface, imparting the stability in the presence of salt. Our study clearly indicate that salt is necessary for maintaining the secondary and tertiary structure of halophilic protein, which is a necessary prerequisite for catalysis. |
| مصطلحات الفهرس: | Biophysics, Biochemistry, Microbiology, Molecular Biology, Biotechnology, 39999 Chemical Sciences not elsewhere classified, 69999 Biological Sciences not elsewhere classified, Inorganic Chemistry, Collection |
| DOI: | 10.6084.M9.FIGSHARE.C.3143491.V1 |
| الاتاحة: | Open access content. Open access content CC-BY http://creativecommons.org/licenses/by/3.0/us |
| Other Numbers: | CNBMS oai:oai.datacite.org:8018885 doi:10.6084/M9.FIGSHARE.C.3143491.V1 947713159 |
| المصدر المساهم: | BEIJING NO.80 MIDDLE SCH From OAIster®, provided by the OCLC Cooperative. |
| رقم الانضمام: | edsoai.ocn947713159 |
| قاعدة البيانات: | OAIster |
| DOI: | 10.6084.M9.FIGSHARE.C.3143491.V1 |
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