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Immobilization of Lipase from Pseudomonas Cepacia and Its Application in the Production of Biodiesel

Author: ZengShuHua
Tutor: ZuoYunJun
School: Huazhong University of Science and Technology
Course: Microbiology
Keywords: Pseudomonas cepacia Lipase Immobilized Enzymatic Properties Transesterification reaction Biodiesel
CLC: Q814.9
Type: Master's thesis
Year: 2007
Downloads: 170
Quote: 1
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Abstract


Lipase immobilization technique helps to improve the stability of the enzyme and to maintain the catalytic activity of the enzyme, the enzyme can be reused in the engineering conditions, thereby reducing the cost of production of the bio-industries. In this paper, tetramethoxysilane (TMOS) and methyl trimethoxysilane lane (MTMS) precursor sol - gel method (sol-gel) fixed Pseudomonas cepacia lipase (Pseudomonas cepacia), optimization the immobilization conditions. The results show that when when the TMOS 2 mmol MTMS 10 mmol, the optimum immobilization conditions: the molar ratio of water with a silane precursor 10, PEG400 120μL, to the amount of enzyme 200 mg. Under these conditions, the immobilized enzyme protein recovery was 93.7% the pNPP hydrolysis specific activity was 1.54 times the free enzyme. On this basis, study the enzymatic properties of immobilized enzyme and free enzyme difference. The results show that, below 80 ° C, the immobilized enzyme can be maintained more than 80% of the enzymatic activity of the free enzyme at 50 ° C or above activity sharply decreased to 80 ° C when the residual activity of about 10%; immobilized enzyme in 50% methanol in Processing 48 h to maintain the enzyme activity of 85% in 90% ethanol for 48 h to maintain 31% of the enzyme activity, while only 69% of the residual activity of the free enzyme and 0; pH stability of the immobilized enzyme and free enzyme also improved to some extent. Determination of enzyme kinetic parameters: Vmax of the immobilized enzyme the 12.3mmol/min mg, Km value of 4.04 mmol / L free enzyme Vmax the 8.0mmol/min mg, Km value 13.64mmol / L, The results showed that the immobilized enzyme is better than the free enzyme substrate affinity and catalytic activity. Then compared derived from the difference of of Japan Amano Enzyme seven free lipase transesterification efficiency, filter out the most efficient transesterification Lipase PS sol - gel method for immobilization of soybean oil catalyzed preparation of biodiesel; system immobilized Lipase PS-catalyzed exchange reaction of soybean oil with methanol ester Synthesis of biodiesel in various influence factors (including the amount of enzyme, the ratio of alcohol to oil, water content, and the reaction temperature, reaction time, solvent, etc.) ester exchange efficiency affected. The results show that, by the ester exchange reaction, catalytic 4.5g soybean oil synthetic biodiesel optimum reaction process parameters: the immobilized enzyme 646mg, alcohol oil ratio of 4:1, 6% moisture content, the reaction temperature of 40 ° C. This condition, the reaction after 72 h, methyl ester highest yield of 96.33%, 16.3 percentage points higher than the free enzyme, and has managed to maintain 60% of the activity of the immobilized enzyme was repeated 10 times.

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CLC: > Biological Sciences > Bioengineering ( Biotechnology ) > Enzyme Engineering > Application of enzymes
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