Immobilized lipase, preparation method thereof and application in catalyzed synthesis of glyceride type PUFA

An immobilized lipase, glyceride type technology, applied in the direction of immobilization on/in organic carriers, fermentation, etc., can solve the problems of low triglyceride content, low enzyme catalysis efficiency, low substrate conversion rate, etc., Achieve the effect of saving production cost and good stability

Active Publication Date: 2016-08-24
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the problems of low enzyme catalytic efficiency, low substrate conversion rate and low triglyceride content in glycerides in the existing preparation method of gl

Method used

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  • Immobilized lipase, preparation method thereof and application in catalyzed synthesis of glyceride type PUFA
  • Immobilized lipase, preparation method thereof and application in catalyzed synthesis of glyceride type PUFA
  • Immobilized lipase, preparation method thereof and application in catalyzed synthesis of glyceride type PUFA

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Experimental program
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Effect test

Embodiment 1

[0025] Preparation of immobilized lipase MAS1

[0026] 1.1 Pretreatment and screening of immobilized resin

[0027] 10g of XAD1180 / DA201 / HP20 / HP2MGL / AB8 five kinds of macroporous adsorption resins were respectively treated with 30mL 95% ethanol, 5% hydrochloric acid and 2% sodium hydroxide for 24h, 4h, 4h respectively. After each step of treatment, the supernatant was filtered Wash it repeatedly with distilled water until the pH value of the supernatant is 7.0, and then proceed to the next step. Finally, the resin is soaked in 20mmol / L buffer solution with different pH values ​​for 4h, and the filtered resin is placed in Store in a 4°C refrigerator for later use. The purpose of soaking the resin with ethanol and acid and alkali is to drive out the air bubbles in the macropores and remove the residual monomers and compounds in the macropores respectively.

[0028] Mix lipase MAS1 enzyme solution 50mg / g resin, equal volume of 20mM pH8 phosphate buffer, and resin pretreated wit...

Embodiment 2

[0041] Synthesis of n-3PUFA-enriched triglycerides through esterification reaction catalyzed by immobilized lipase MAS1

[0042] 10g of substrate (molar ratio of glycerol to PUFA is 1:1, 1:2, 1:3, 1:4, 1:5) was placed in a 250mL Erlenmeyer flask with a stopper, and a certain amount of lipase was added according to the enzyme activity unit ( 750U, 1125U, 1500U, 1875U, 2250U), placed in constant temperature oscillators at different temperatures (50°C, 55°C, 60°C, 65°C, 70°C) for 24 hours of vacuum reaction at a speed of 200rpm, at different reaction times Sampling, HPLC analysis of the composition of the reaction mixture, GC analysis of the fatty acid composition of TAG, and comparison with the esterification reaction catalyzed by Novozym 435 with the same enzyme activity unit under the same conditions.

[0043] Depend on figure 2 (a) It can be seen that under the condition that the molar ratio of glycerol to n-3PUFA is 1:3 and 750 U of immobilized enzyme, when the temperature...

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Abstract

The invention discloses an immobilized lipase, a preparation method thereof and application in catalyzed synthesis of glyceride type PUFA. The lipase is from lipase MAS1 of Streptomyces, and an immobilized carrier is XAD1180 resin. The preparation method of the immobilized lipase includes the following steps that 1, the XAD1180 resin is soaked with ethanol, acid and alkali in sequence to remove air bubbles and residues in resin macropores; 2, the lipase MAS1 and the treated XAD1180 resin are mixed in the proportion of 25-150 mg/g resin, and standing is performed to obtain the immobilized lipase. The immobilized lipase has better stability and repeatability compared with free lipase, and the production costs are saved. In addition, the esterification efficiency is up to 99.31%, and the PUFA triglyceride content is up to 92.26%.

Description

technical field [0001] The invention relates to the preparation of immobilized lipase and its application technology in catalytically synthesizing high-content glyceride type PUFA. Background technique [0002] Polyunsaturated fatty acids (PUFA) generally refer to straight-chain fatty acids containing two or more double bonds and a carbon chain length of 18 to 22 carbon atoms, mainly derived from marine fish oils and vegetable oils. Current studies have shown that PUFAs with important physiological activities mainly include eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), eicosapentaenoic acid (DPA), arachidonic acid (AA), Stearidonic acid (SDA), α-linolenic acid (ALA), γ-linolenic acid (GLA), etc. It has physiological regulation functions such as reducing blood viscosity of human body, improving blood microcirculation, lowering blood cholesterol and triglyceride, etc., and is currently receiving increasing attention. [0003] Compared with other forms of PUFA (mai...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12P7/64
CPCC12N11/08C12P7/6454C12P7/6472
Inventor 王永华蓝东明汪秀妹杨博王卫飞
Owner SOUTH CHINA UNIV OF TECH
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