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Fur-Grip gold wire fruiting bodies stipe cell wall extension growth studies
Author: LiJiYing
Tutor: YuanSheng
School: Nanjing Normal University
Course: Microorganism
Keywords: Flammulina Cell wall Stipe elongation Elongation protein beta - 1 , 3 - glucanase
CLC: Q942.5
Type: Master's thesis
Year: 2005
Downloads: 42
Quote: 0
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Abstract
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In vitro from recombinant heat-inactivated cell wall elongation measurement method from the crude enzyme preparation the various rich in polysaccharide hydrolase screened a cell wall in the hair handle money bacteria ( mushroom , Flammulina velutipes) Stipe elongation protein , this protein can cause cell wall elongation Junbing . Filter crude enzyme preparation through a Bio-Gel P - 60 molecular sieve chromatography , DEAE - Cellulose ion- exchange chromatography and high performance liquid chromatography Synchropak S300 cation exchange chromatography to obtain electrophoretically pure molecular weight of about 21KD protein , this protein added to the cell wall extension analyzer chamber solution instantly cause the rapid elongation of the stipe cell wall . The protein does not contain chitinase activity , the activity of beta - glucan has not found xyloglucan and glucomannan had no role in the use of Chinese Academy of Sciences Shanghai Institute of Biochemistry Proteomics Analysis Center LCQ-DECA-XP mass spectrometry of the protein sequence analysis , the results show that the the existing protein library does not match the fragment to Edman and LS / MS / MS are two ways sequencing ; Harvard University, the protein by trypsin cleavage , get two a peptide fragment , respectively, to the amino acid sequence of the seven EFHWAVPK the peptides and MFSNDVLAR nonapeptide , GenBank did not find the same protein sequence , and also found no matching peptides , presumably as a new protein . Additionally discovered a new class of beta -1 , 3 - glucanase, in SDS-PAGE showed a molecular weight of 33kD electrophoretic bands in the screening process . This protein hydrolase activity but there is a strong hydrolase activity in the barley beta - glucan , microcrystalline cellulose , it does not have to cause the the stipe cell walls elongated function . The characteristics of the study show that the optimum temperature and optimum pH of the enzyme at 37 ° C and 4.5 , respectively , at pH lower than 4.0 when the enzyme is more stable . Not found an exact match with the peptides by mass spectrometry LCQ-DECA-XP peptide mapping analysis , presumably for a new inner cut beta - 1 , 3 - glucanase .
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CLC: > Biological Sciences > Botany > Plant cytology > Plant Cell Physiology
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