Method for enriching n-3 polyunsaturated fatty acids by enzymic method

Abstract

The invention discloses a method for enriching n-3 polyunsaturated fatty acids by an enzymic method, and belongs to the field of grease deep processing. The method disclosed by the invention comprisesthe steps that: grease, a water (alcohol) solution and lipase are taken to react in a reactor, wherein the lipase comprises one or more kinds of lipase from Candida cylindracea and lipase Candida antarctica lipase A; and the adding sequence of the lipase is uncertain. At present, the content of saturated and monounsaturated fatty acids in glyceride obtained by enzymatic catalytic hydrolysis or alcoholysis PUFA enrichment reaction is still high; two kinds of lipase from different sources are selected and have specificity in hydrolysis or alcoholysis of saturated fatty acids and monounsaturatedfatty acids in the invention; the fatty acid content of non-PUFA in a glyceride product is further reduced; and therefore, the glyceride product with higher PUFA content is obtained.

Description

technical field

[0001] The invention relates to a method for enzymatically enriching n-3 polyunsaturated fatty acids, which belongs to the field of oil deep processing. Background technique

[0002] Polyunsaturated fatty acid (PUFA) has important biological significance to the human body, especially n-3PUFA, which has been widely reported in the world. Among them, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) can significantly prevent and treat cardiovascular diseases, assist in inflammation repair, and promote the normal development of infant sensory organs, and are widely used in health food and medicines.

[0003] The n-3PUFAs products on the market mainly have three forms: ethyl ester type, glyceride type and free fatty acid type. The content of n-3PUFAs in the glyceride type is relatively low, about 30%, which cannot meet people's health care and medicinal purposes. need. The content of n-3PUFAs in the ethyl ester type and free fatty acid type can reach ...

Examples

Embodiment 1

[0043] Accurately weigh the fish oil (the fatty acid composition of the fish oil sample is as follows: figure 1 Shown, wherein n-3PUFA=34.3%, SFA%=36.3%, MUFA%=24.0%) 3.0g, phosphate buffer solution 3g (concentration is 0.1mol / L, pH is 7) and AY "Amano" 400SD lipase 960U, added to the reaction kettle, put into the magnetic rotor, and sealed. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After the reaction, use KOH-ethanol aqueous solution to remove the hydrolyzed free fatty acid, then wash it with water three times, take the upper clear oil phase, and evaporate the solvent to obtain fish oil glycerides rich in n-3PUFAs (fatty acid composition such as figure 2 ).

[0044] Accurately weigh 10.0g of the obtained glyceride (the product accumulated many times in the previous step), 5g of ethanol solution (alcohol-water mass rati...

Embodiment 2

[0046] Accurately weigh 10.0 g of fish oil, 5 g of ethanol solution (alcohol-water mass ratio 1:4) and 4500 U of Candida antarcticalipase A lipase, add them into the reaction kettle, put it into a magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After reaction finishes, molecular distillation removes free fatty acid and ethyl ester in the product, obtains the fish oil glyceride that is rich in n-3PUFAs (fatty acid composition such as Figure 4 ).

[0047] Accurately weigh 3.0 g of glyceride, 3 g of phosphate buffer solution and 960 U of AY "Amano" 400SD lipase, add them into the reaction kettle, put it into the magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant ...

Embodiment 3

[0049] Accurately weigh 3.0g of fish oil, 3.0g of phosphate buffer solution, 960U of AY "Amano" 400SD lipase and 1350U of Candidaantarctica lipase A lipase, add them into the reaction kettle, put it into the magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After the reaction, use KOH-ethanol aqueous solution to remove the hydrolyzed free fatty acid, then wash with water three times, take the upper clear oil phase, evaporate the solvent to obtain fish oil glyceride rich in n-3PUFA. In the fatty acid composition of glyceride products, the content changes of n-3PUFA, SFA and MUFA are shown in Table 1.

[0050] The experimental conditions and results of Examples 4 to 19 are shown in Table 1 below, wherein, except for the conditions indicated, the remaining operating parameters of Examples 4, 6, 8, 11, 12,...

technical field

[0001] The invention relates to a method for enzymatically enriching n-3 polyunsaturated fatty acids, which belongs to the field of oil deep processing. Background technique

[0002] Polyunsaturated fatty acid (PUFA) has important biological significance to the human body, especially n-3PUFA, which has been widely reported in the world. Among them, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) can significantly prevent and treat cardiovascular diseases, assist in inflammation repair, and promote the normal development of infant sensory organs, and are widely used in health food and medicines.

[0003] The n-3PUFAs products on the market mainly have three forms: ethyl ester type, glyceride type and free fatty acid type. The content of n-3PUFAs in the glyceride type is relatively low, about 30%, which cannot meet people's health care and medicinal purposes. need. The content of n-3PUFAs in the ethyl ester type and free fatty acid type can reach ...

Examples

Embodiment 1

[0043] Accurately weigh the fish oil (the fatty acid composition of the fish oil sample is as follows: figure 1 Shown, wherein n-3PUFA=34.3%, SFA%=36.3%, MUFA%=24.0%) 3.0g, phosphate buffer solution 3g (concentration is 0.1mol / L, pH is 7) and AY "Amano" 400SD lipase 960U, added to the reaction kettle, put into the magnetic rotor, and sealed. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After the reaction, use KOH-ethanol aqueous solution to remove the hydrolyzed free fatty acid, then wash it with water three times, take the upper clear oil phase, and evaporate the solvent to obtain fish oil glycerides rich in n-3PUFAs (fatty acid composition such as figure 2 ).

[0044] Accurately weigh 10.0g of the obtained glyceride (the product accumulated many times in the previous step), 5g of ethanol solution (alcohol-water mass rati...

Embodiment 2

[0046] Accurately weigh 10.0 g of fish oil, 5 g of ethanol solution (alcohol-water mass ratio 1:4) and 4500 U of Candida antarcticalipase A lipase, add them into the reaction kettle, put it into a magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After reaction finishes, molecular distillation removes free fatty acid and ethyl ester in the product, obtains the fish oil glyceride that is rich in n-3PUFAs (fatty acid composition such as Figure 4 ).

[0047] Accurately weigh 3.0 g of glyceride, 3 g of phosphate buffer solution and 960 U of AY "Amano" 400SD lipase, add them into the reaction kettle, put it into the magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant ...

Embodiment 3

[0049] Accurately weigh 3.0g of fish oil, 3.0g of phosphate buffer solution, 960U of AY "Amano" 400SD lipase and 1350U of Candidaantarctica lipase A lipase, add them into the reaction kettle, put it into the magnetic rotor, and seal it. The reaction kettle was placed on a magnetic stirrer, and the reaction kettle was connected to circulating water, and the water temperature was kept at a constant temperature of 37° C. for 10 hours. After the reaction, use KOH-ethanol aqueous solution to remove the hydrolyzed free fatty acid, then wash with water three times, take the upper clear oil phase, evaporate the solvent to obtain fish oil glyceride rich in n-3PUFA. In the fatty acid composition of glyceride products, the content changes of n-3PUFA, SFA and MUFA are shown in Table 1.

[0050] The experimental conditions and results of Examples 4 to 19 are shown in Table 1 below, wherein, except for the conditions indicated, the remaining operating parameters of Examples 4, 6, 8, 11, 12,...

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