Please use this identifier to cite or link to this item: http://hdl.handle.net/10603/193629
Title: Studies on Amylase and Amylase Inhibitors of Tribolium Spp
Researcher: Khillare Neeta Sudamrao
Guide(s): Kamble L. H.
Keywords: n.a.
University: Swami Ramanand Teerth Marathwada University
Completed Date: 24/12/2016
Abstract: Amylase inhibitors (AIs) play an important role in host seed resistance to insect newlinepests. Therefore, the present studies were carried out on seed amylase inhibitors for newlineinhibition of amylase and their biological activity against the stored product insect, newlineTribolium castaneum (Herbst), the most important pest of stored products distributed newlinethroughout the world. newlineThe present study indicated that amylase was extracted from larvae of T. newlinecastaneum, partially purified and characterized using substrate assays. Starch was used as newlinea substrate to detect electrophoretically separated amylase of T. castaneum larvae extract newlineon Native-PAGE and SDS-PAGE. In gel activation method involved electrophoresis, newlinefollowed by washing the gel with water in case of Native-PAGE, equilibration of the gel newlinein starch (1%) buffer, followed by washing with water to remove hydrolysed starch, newlinerevealing amylase bands. newlineTribolium castaneum larvae amylase showed activity over a broad range of pH, newlinemaximum activity was observed between the pH 5.8-7.0. Larvae amylase showed optimal newlineactivity at 40 ºC. Although larvae amylase activation was affected by Na+, Mg2+ , Ba2+, newlineK+ and Zn2+ While Mn2+significantly increased activity. newlineThe T. castaneum larvae amylase isoform was observed in the fraction eluted on newlineDEAE-Cellulose column and activation measured by substrate assays. Purification and newlinesubstrate assay studies revealed that, 38.73kDa polypeptide was amylase protein. newlineThe molecular mass was analyzed by MALDI-TOF to detect the highly accurate newlinemolecular weight information on intact molecules. The ability to generate such accurate newlineinformation can be extremely useful for protein identification and characterization. The newlineobtained spectrum of m/z ratio verses intensity showed a single peak having molecular newlinemass 37.69Kda. newlineThe single polypeptide extracted and purified from T. castaneum larvae was newlinetryptic digested and the peptides were determined by TOF/Q-TOF Mass spectrometer. newlineThere were 16 peptides analyzed on spectrometer in m/z ratio 300-3200da.These peaks newlinewere having
Pagination: 108p
URI: http://hdl.handle.net/10603/193629
Appears in Departments:School of Life Sciences

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01_title.pdfAttached File68.81 kBAdobe PDFView/Open
02_certificate.pdf132.13 kBAdobe PDFView/Open
03_abstract.pdf60.69 kBAdobe PDFView/Open
04_declaration.pdf70.4 kBAdobe PDFView/Open
05_acknowledgement.pdf17.82 kBAdobe PDFView/Open
06_contents.pdf56.79 kBAdobe PDFView/Open
07_list_of_tables.pdf32.53 kBAdobe PDFView/Open
08_list_of_figures.pdf53.73 kBAdobe PDFView/Open
09_abbreviations.pdf32.1 kBAdobe PDFView/Open
10_chapter1.pdf2.74 MBAdobe PDFView/Open
11_chapter2.pdf142.53 kBAdobe PDFView/Open
12_chapter3.pdf170.53 kBAdobe PDFView/Open
13_chapter4.pdf14.39 MBAdobe PDFView/Open
14_conclusion.pdf52.9 kBAdobe PDFView/Open
15_bibliography.pdf149.04 kBAdobe PDFView/Open


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