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Isolation and Identification of Highly-productive Cellulase Strain and Study on the Condition for Liquid-state Fermentation

Author: LiuHaiBo
Tutor: WangYiQiang;ChenJieNan
School: Central South University of Forestry Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Cellulase Filter Appraisal Liquid fermentation Optimization
CLC: TQ925
Type: Master's thesis
Year: 2008
Downloads: 152
Quote: 0
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Abstract


Cellulose is the earth's most abundant, the most extensive source of organic matter is an important class of renewable resources and energy. Due to the special nature of the molecular structure of cellulose, and cellulose material in the existence of the state of nature particularity, so that these valuable resources is difficult to be rational utilization. A large number of studies have shown that the biodegradation of cellulose has incomparable advantages of the other methods. To make the cellulose is broken down into small molecules or other nutrients, screening the excellent bacteria isolated from the conversion of cellulose to rapid degradation. To spaces of the perennial accumulation cellulose waste, dead rotting trees in the forest and paper waste the underwater sludge as good bacteria sources, and collect samples. Cellulose Congo red medium plate culture stained of transparent hydrolytic ring size and the size of the colony diameter ratio, the screening of the medium for the process of filter paper cellulose filter paper crash speed and the filter paper weightlessness rate, determination of cellulase activity rescreening , Isolation of the mold to a plant height of cellulase activity - ZN-205 combination of morphological characteristics observed and 18s rRNA gene sequence analysis to determine the strain Java Eupenicillium (Eupenicillium javanicum). A cellulose liquid fermentation culture conditions and fermentation medium components on the Java Eupenicillium (Eupenicillium javanicumZN-205 enzyme production. Experimental research conditions in the papers, experimental: medium best carbon source straw powder, the best nitrogen source KNO3, plus a certain percentage of bran significantly increased the enzyme activity in the fermentation system, and add the appropriate concentration of surfactant no significant role in the improvement of enzyme production capacity. the orthogonal optimization test results show that, The fermentation broth 100mL, 250mL flask Zhongzhuang medium initial pH of 5.5 KNO3 added concentration of 1.0%, straw powder and wheat bran ratio of 7:3, Twain dosage of 0.4% at a temperature of 30 ° C, shaking speed 175r/min cultured under conditions 5d strain has maximum enzyme production capacity, which carboxymethyl cellulose (CMC) activity 24.52IU/mL filter paper activity (FPA) for 2.68IU/mL on Java Eupenicillium (of Eupenicillium javanicunZN 205 generated extracellular enzyme extraction methods, the enzymatic reaction conditions of the study results showed that: with 3000r/min.15min centrifugation conditions to obtain a crude enzyme solution, CMC highest activity, which enzyme promote optimal reaction temperature is 60 ℃, the optimum pH value of 5.0; crude enzyme solution with 4000r/min.10min centrifugation conditions, FPA highest, its enzymatic reaction optimum temperature of 50 ° C, the optimum pH value of 5.0. , CMC enzyme and filter enzyme stability, good enzyme stability of the CMC enzyme is between pH 4-5, at a pH less than 3 and greater than 6:00, the stability of the enzyme are sharply; in 40 ° C and 50 ° C and 60 ℃ enzyme thermal stability is better, rather than a sharp decline in the stability of the enzyme in 60 ℃. filter paper enzyme at pH 5-6 between enzyme stability is better, while when the pH is greater than 7 and less than 4:00 enzyme activity sharply; at 40 ° C, 50 ° C when thermal stability is better, while the enzyme activity at 60 ° C, 70 ° C for a sharp decline, poor stability of the enzyme. concentration of 1.0mmol / L, Na, K, Fe2, Mg2 activation, while Zn2, Cu2, Ca2, Mn2 activity of the CMC and filter paper activity of inhibitory activity of the CMC and filter paper activity.

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