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Method for synthesizing biodiesel by two-step catalysis of lipid via Yarrowia lipolytica lipase, and biodiesel obtained thereby

A biodiesel and yeast technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of increasing the risk of industrialized production, increasing the intensity of equipment investment, and affecting production efficiency, and achieving increased industrialization feasibility. performance, improved production efficiency, and extended service life

Inactive Publication Date: 2009-04-22
领先生物农业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the organic solvents used in the reaction system have a very low boiling range (such as petroleum ether, tert-butanol, with a boiling range of 40-90° C.), are flammable and explosive, increase the risk of industrial production, and increase the input intensity of equipment. , and its production efficiency is affected due to the use of a large amount of solvent

Method used

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  • Method for synthesizing biodiesel by two-step catalysis of lipid via Yarrowia lipolytica lipase, and biodiesel obtained thereby
  • Method for synthesizing biodiesel by two-step catalysis of lipid via Yarrowia lipolytica lipase, and biodiesel obtained thereby
  • Method for synthesizing biodiesel by two-step catalysis of lipid via Yarrowia lipolytica lipase, and biodiesel obtained thereby

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Preparation of crude lipase enzyme solution: Yarrowia lipplytica CGMCCNO.2707 was cultured on the slope of eggplant bottle for 12 hours and then inoculated in a fermenter for cultivation. After 90 hours of cultivation, the hydrolase activity was measured to reach 8500u / ml Lower the tank, collect the fermentation broth after the lower tank, use the model SSP450 centrifuge produced by Zhangjiagang Xinhua Chemical Machinery Co., Ltd. to centrifuge at 1900 rpm, and take out the solids and bacterial cells in the fermentation broth to obtain the crude Lipase enzyme solution.

[0061] Preparation of immobilized lipase: based on the total weight of lipase fermentation broth, first mix 2‰ polyvinyl alcohol, 1‰ Tween 80, 2‰ PVP and 1‰ MgSO 4 The immobilized auxiliary agent is added to the lipase fermentation broth and mixed evenly. Mix silk (about 1:1.5, the ratio of the weight of the carrier to the crude lipase enzyme solution) with the crude lipase enzyme solution, so that the amou...

Embodiment 2

[0065] The same as in Example 1, except that 20g of commercially available soybean oil was added to a 100ml ground-mouthed conical flask with a stopper, and then 12ml (enzyme activity 480U / ml) of Candida rugosa 14830 lipase fermentation broth was added. The reaction is carried out by rpm incubation and shaking. The method for preparing fatty acids in Example 1 is used to obtain the crude fatty acids used for the esterification reaction.

[0066] Take 1.39g of the above fatty acid, add 291ul of ethanol, add 0.14g of the immobilized lipase in Example 1, and react with shaking at 30°C and 190 rpm for 8 hours. The final fatty acid ethyl ester conversion rate is measured by the acid value analysis method. Up to 81%.

Embodiment 3

[0068] The same as in Example 1, except that 20g of soybean acidified oil purchased from Qinhuangdao Jinhai Cereals, Oils and Foodstuffs Co., Ltd. was added into a 100ml ground-mouthed conical flask, and then 0.4ml (enzyme activity 5000U / ml) lipolytic Yarrow was added. Yeast strain CGMCC NO.2707 crude lipase enzyme solution, and then adopt the method of preparing fatty acid in Example 1 at 40°C, and react with 130 rpm incubation and shaking to obtain crude fatty acid used for esterification reaction.

[0069] Take 1.40g of the fatty acid described in Example 1, add 202ul of methanol, add 0.14g of the immobilized enzyme described in Example 1, and shake the reaction at 30°C and 190 rpm for 8 hours, using the detection method in Example 1. The final fatty acid methyl ester conversion rate reached 82%.

[0070] The obtained certain amount of fatty acid methyl esters are subjected to vacuum distillation under a vacuum degree of 95KPa, and the fractions at 190-210°C are collected as th...

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Abstract

The present invention provides a crude lipase enzyme solution of Yarrowia lipolytica microzyme CGMCC No.2707 and a method for synthesizing biological diesel oil by catalyzing the hydrolization and esterification reaction of waste animal and vegetable fat by the immobilized lipase of the crude lipase enzyme solution. A method for preparing the lipase crude enzyme solution and the immobilized enzyme is also provided. The method uses hydrolization and esterification reactions of the oil and fat use lipase as catalysts, but does not use strong acid and strong base. The method has the advantages of mild reaction condition and small pollution. Organic solvent is not used during the entire production process of the fatty acid methyl ester / ethyl ester. The reaction is safe and the efficiency is high.

Description

【Technical Field】 [0001] The invention belongs to the field of biochemical technology. More specifically, the present invention relates to a method for synthesizing biodiesel from fats and oils using biological enzymes and the obtained diesel. 【Background technique】 [0002] The combustion performance of biodiesel is comparable to that of petrochemical diesel. It is a new type of pollution-free renewable energy. Its research and application have received extensive attention. [0003] The current method of producing biodiesel is mainly chemical method, which uses high temperature and high pressure, high reaction energy consumption, many side reactions, and strict raw material requirements. Research on the green chemical method of bio-enzymatic synthesis of biodiesel is becoming a hot spot in the biodiesel industry. Enzymatic synthesis of biodiesel generally has many advantages such as mild reaction conditions, fewer side reactions, and no pollutant emissions during the reaction. ...

Claims

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Application Information

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IPC IPC(8): C12N9/20C12P7/64C12R1/645
CPCY02E50/13Y02E50/10
Inventor 孟永宏徐志文李月娟周志奇梁晓梅杨娜陈晋南
Owner 领先生物农业股份有限公司
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