Cellulase synthesis in 2-deoxy glucose resistant mutants of trichoderma reesei p-2
KrishiKosh
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Title |
Cellulase synthesis in 2-deoxy glucose resistant mutants of trichoderma reesei p-2
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Creator |
Tina Mittra
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Contributor |
Chaudhary, Kamla
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Subject |
Enzymes, Mutants, Fungi, Cellulose, Productivity, Proteins, Electrophoresis, Solutes, Concentrates, Oligosaccharides
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Description |
A study was under taken to isolate 2-DG resistant mutants of Trichoderma reesei P-2 for cellulase synthesis. Spores of T. reesei were mutagenized with UV irradiation for a time period of 15 minutes. At this time maximum number of mutants were obtained.Antimetabolite resistant mutants were obtained on 0.05% 2-DG containing Reese medium plates. A total of 24 isolates which grew profusely, were purified and again patched on Reese medium contained 2-DG. Finally 11 mutants showed good growth on this medium. 2-DG resistant mutants of T. reesei mutants MU-22 and MU-24 showed maximum cellulase production and selected. T.reesei mutants MU-22 and MU-24 showed maximum FPase activity (1.54 IU/ml and 1.56 IU/ml) respectively, CMCase activity (6.4IU/ml and 5.5IU/ml) respectively in cellulose medium. However and -glucosidase activity was more in MU-22(0.28 IU/ml). Effect of different carbon sources on synthesis of cellulases were analysed in mutants MU-24 and MU-22. Cellulose was found to be the best carbon source whereas in cellobiose and cotton medium, cellulase activity was reduced. In phosphocellulose medium -glucosidase activity was high and sorbitol was not a good inducer for the synthesis of cellulases. The optimization studies for cellulase production by MU-24 showed that cellulose at 1% concentration produced high level of cellulases. Similarly ammonium sulphate at a concentration of 0.14% was found to be essential for all the three enzyme activity. Since T. reesei is a mesophilic organism, at high temperature it did not grow. The optimum temperature for growth of T. reesei was found to be 30ºC. Saccharification of rice straw indicated the release of 50% reducing sugar which were more (35%) then the parents strain. The protein profile of culture filtrate of T. reesei mutants were determined by SDS PAGE. It was observed that the first set of proteins which appeared during cellulase induction were of 50KDa, 40KDa, 30KDa and 25KDa. Thus from this investigation it can be concluded that level of exoglucanase, endoglucanase and -glucosidase has been altered by mutation and such improvement in altering the level of individual component of cellulase complex is practically possible. |
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Date |
2016-12-24T09:49:00Z
2016-12-24T09:49:00Z 2005 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/92572
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Language |
en
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Format |
application/pdf
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Publisher |
CCSHAU
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