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Sophorose the extraction and purification for the inter- type Neurospora cellulase produced induction of

Author: DengXiaoHong
Tutor: ChenYuRu
School: Nanjing Normal University
Course: Microbiology
Keywords: Neurospora intermedia cellulose sophorose abduction fermentation
CLC: TQ925
Type: Master's thesis
Year: 2011
Downloads: 15
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It was studied that the optimum conditions of solid fermentation of Neurospora intermedia to produce cellulose through orthogonal test and single factor experiments and the activities of CMCase,β-glucosidase and FPAase were measured.The activities of CMCase and B-glucosidase were used as main reference indexs.The results showed that the components of the optimal solid culture followed the following ratio of matter: bran to rice husk (1:1);solid materials to water (1:1.5); (NH4)2SO43% and that the optimal conditions for cellulose production are that initial pH was 7.0, culture temperature 34℃and cultivating time 108h. The activities of CMCase, FPAase andβ-glucosidase are respectively 668.0U/g、162.5U/g and 939.5U/g under these optimal zymotic conditions. The enzyme showed optimal activity at 55-60℃and pH 4.5-5.0. The glucose and cane sugar can be hydrolyzed and isomeried visibly by Neurospora intermedia solid enzymes.Cellulase production from liquid mixed fermentation of Neurospora intermedia and Trichodermas viride was investigated. It was found that the largest CMCase activity value,96.3U/mL, was obtained when Trichodermas viride delay in Neurospora intermedia was 24h and volumetric inoculum ratio of Neurospora intermedia versus Trichodermas viride was 5:1, higher than their’s of the monoculture of them. As for FPAase activity,22.8U/mL, was postponed for 1 day to come maximal enzyme activity, higher than their’s of the monoculture of them.The extraction and purification technique of sophorose from crude stevioside is studied in this paper. The article was focus on the conditions of extraction and purification of sophorose from crude steviosides. Several factors, such as concentration of crude stevioside, concentration of acid, type of acid and hydrolysis time. The optimal conditions are optimized by the method of orthogonal design, and the optimum condition is determined. The optimal conditions were as follows: concentration of crude steviosides for 1%, concentration of sulfuric acid for 0.3N, hydrolysis time for 60min, and the hydrolysis yield was 34.1%. Chromatographed by activated carbon column, most impurities were removed. The target products were identified for sophorose by LC-MS.The preliminary separation and extraction of flavonoids from from sophora japonica was studied. The purity of total flavonoids and isoflavones from crude extracts are 61.3% and 21.6%. The sophorose content of sophora japonica is 0.52% when sophora japonica was boiled by water. Kaempferol, sophoricoside, genistin and genistein were identified.Cellulase inducement from sphorose was studied. It was found that the time of the maximal enzyme activity, the CMCase activity,46.3U/mL,24th, was gained when the sophorose concentration was 10-2M. The time point of adding abduction including sophorose for cellulose production affects the positive impact for eyzyme output and activity. The CMCase activity keeps a low and changleless almostly value along with the sophorose concentration elevatory, when the CMCase activity value was measured at 3th. When the CMCase activity value was measured at 18th and 24th, They get more and more substantial along with increasing of concentration of sophorose; The maximal value respectively,24.7U/mL and 68.0 U/mL at 10-1M. It is proved that the ability of sophorose for clllulase production correspond with certain time factor. Whether different hours of contacting Neurospora hypha with sophorose have an effect on the maximal enzyme activity was studied, it was found that the enzyme activity value at 6th,12th and 18th when the abduction including sophorose was filtered out, were 22.1%,43.4% and 63.7% respectively comparing with control experiment which the sophorose abduction was reserved.

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CLC: > Industrial Technology > Chemical Industry > Other chemical industries > Fermentation industry > Enzyme preparation ( enzyme )
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