The triglyceride Production by Enzymes method of the unsaturated mono-glycerides of a kind of coproduction
Technical field
The present invention relates to the unsaturated mono-glycerides of a kind of coproduction and triglyceride Production by Enzymes method.
Background technology
Natural food oils main component is triglyceride level, and triglyceride level is that a glycerine is in conjunction with the compound of three fatty acid molecules.In natural fats and oils, also have a small amount of triglyceride, triglyceride is on a glycerol molecule, to be connected with the compound that two molecules of fatty acids form, the common content of the content of triglyceride in edible oil is no more than 5%.1993, Hara K reported first triglyceride there is function (the Ann Nutr Metab1993 that reduces blood fat; 37:185), prompting can for prevention and treatment hyperlipidaemia and with the closely-related cardiovascular and cerebrovascular diseases of hyperlipidaemia.Japanese Kao company has released triglyceride type edible oil first in later 1990s, and obtains coml success.Afterwards, increasing research shows that triglyceride has the multiple function that is conducive to HUMAN HEALTH, and experimentation on animals and human body all prove that triglyceride has unique metabolic patterns, adopts triglyceride to substitute triglyceride level, can prevention of obesity, reduce blood fat.
Triglyceride as edible oil is generally unsaturated type triglyceride, and it is generally to adopt enzymatic conversion method explained hereafter.More representative production technique is first triglyceride level to be decomposed and obtains free fatty acids, then to adopt lipase be that catalysis instrument makes the reaction of free fatty acids and glycerine esterification obtain triglyceride.In this technique, the acquisition of lipid acid can adopt highly pressured hydrolysis, also can adopt enzymatic hydrolysis.In the enzyme process building-up process of triglyceride, all follow a certain amount of mono-glycerides to generate, mono-glycerides also has very high using value, still, and when adopting lipid acid to be substrate synthetic triglyceride, still there are a considerable amount of lipid acid residues, when fractionation by distillation, very difficult that lipid acid is thoroughly separated with mono-glycerides, so, be difficult to obtain the mono-glycerides product of low acid value, and the application of the mono-glycerides of high acid value is very limited.
Glycerine solution technique is the another kind of important method of producing triglyceride, the method be take triglyceride level as substrate, in reaction process, do not need to add free fatty acids, therefore, avoid lipid acid in above-mentioned technique to be difficult to separated problem with mono-glycerides, when producing triglyceride, obtained the mono-glycerides product that acid value is lower.In these class methods, because glycerine has package action to enzyme, for addressing this problem, patent CN200310112327.X adopts the glycerine of carrier adsorption form as substrate, and reaction is carried out.Practice is found, although the glycerine of adsorption form can overcome the defect that free glycerol is difficult to reaction, but still exist, reaction efficiency is low regenerates and use inconvenient shortcoming with carrier, and the difficulty of industrialized implementation is very large.
Summary of the invention
The object of the invention is to the shortcoming existing for prior art, the triglyceride Production by Enzymes method of the unsaturated mono-glycerides of a kind of coproduction is provided.It is substrate that the method adopts triglyceride type grease, first carries out ethanolysis reaction, obtains reaction product, adds free glycerol in reaction product, carries out enzyme process glycerolysis reaction, can when obtaining triglyceride, obtain the unsaturated mono-glycerides of low acid value.
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
(1) in natural fats and oils, add dehydrated alcohol, under fixed lipase catalyzed, carry out ethanolysis reaction, making fatty-acid ethyl ester production rate is 5~40%w/w, removes residue ethanol, obtains alcoholysis product;
(2) in step (1) alcoholysis product, add the glycerine of 5~15%w/w, then by above-mentioned mixed substrates in the packed bed enzyme reactor of immobilized lipase is housed, under vacuum condition, react;
(3) step (2) reaction product is through molecular distillation, and the high temperature cut that separation obtains is the product containing triglyceride, obtains take mono-glycerides as main low temperature fraction simultaneously.
The described reaction of step (2) adopts pressure segmentation to control, and under vacuum condition, before reaction, first under normal pressure, reacts certain hour; Wherein the synthesis under normal pressure time length is 1/4~1/3 of the vacuum reaction time.
The condition of the described vacuum reaction of step (2): pounds per square inch absolute (psia) is 10~1000pa.
The described temperature of reaction of step (2) is 50 ℃~60 ℃.
The add-on of the described glycerine of step (2) is 8~12%w/w.
The consumption of the described immobilized lipase of step (2) is 0.2~5% of substrate weight, and the vacuum reaction time length is 3~24 hours.
The described molecular distillation of step (3) adopts three-stage distillation, the degassed and de-glycerine of the first step, and distillation temperature is 80~120 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 150~180 ℃; Third stage separation obtains target product, and distillation temperature is 190~220 ℃.
Immobilized lipase of the present invention is one or more the mixture deriving from root enzyme genus, Aspergillus, hair enzyme genus, bacterium, yeast and steapsase.Be preferably lipase Novozym435, Lipozyme RM IM or Lipozyme TL IM.
Described natural fats and oils is animal grease or Vegetable oil lipoprotein.
Described Vegetable oil lipoprotein is one or more the mixture in soybean oil, rapeseed oil, Semen Maydis oil, plam oil, peanut oil, sunflower seed oil, Camellia oil, sweet oil and Rice pollard oil; Animal grease is one or more the mixture in fish oil, lard, butter and chicken fat.
Step of the present invention (1) is the enzyme process ethanolysis reaction of grease, and reaction product is fatty-acid ethyl ester.The enzyme process ethanolysis of grease is process simply individual and that be easy to realize.The ethanolysis of grease normally mixes dehydrated alcohol and grease, then adds immobilized lipase to stir, or by reaction substrate by the pillar of immobilized lipase is housed, reaction certain hour, can obtain reaction product.The enzyme process ethanolysis temperature of reaction of grease can be set flexibly according to the tolerable temperature of lipase, is generally 20~60 ℃; The addition of reaction times and enzyme is relevant.In the present invention, the add-on by reasonable computation ethanol and control extent of reaction, the production rate of controlling ethyl ester is 5~40%(w/w), in reaction product, the amount of ethyl ester accounts for 5~40%(w/w of reaction product).
Step of the present invention (2) is glycerolysis reaction, is committed step of the present invention.As previously mentioned, the invention is characterized in and adopt free glycerol to carry out glycerine solution.In the present invention, directly in step (1) reaction product, add 5~15%(w/w) glycerine, then by this reaction substrate by the packed bed enzyme reactor of immobilized lipase is housed, under the vacuum condition that catalysis is 10~1000pa in absolute pressure, react.In this step, the addition of glycerine is 5~15%(w/w), glycerine addition is 5~15% of step (1) reaction product weight, preferably 8~12%.Very few glycerine addition, can affect the growing amount of partial glyceride (partial glyceride is the general designation of triglyceride and mono-glycerides), and too much glycerine addition is unnecessary, and superfluous glycerine still tends to be gathered in zymin top layer, affects speed of response.In the present invention, the interpolation of glycerine can be once to add, the mode that also can take stream to add, and the mode that adds of preferred streams.Step of the present invention (2) is particularly limited and will adopts filling bed type enzyme reactor, and with respect to stirring-type enzyme reactor in batches, filling bed type enzyme reactor has better substrate mass-transfer performance, more easily realizes the efficient catalytic of enzyme.The feature of step of the present invention (2) reaction is also that catalyzed reaction is to carry out under the vacuum condition that is 10~1000pa in absolute pressure, vacuum condition can remove the residual ethanol of reaction substrate and micro-moisture, in addition, vacuum condition also impels part ethyl ester in reaction system to be converted into triglyceride.In the present invention, the pressure-controlling of step (2) reaction also can be carried out in segmentation, is first synthesis under normal pressure, carries out after certain hour (be about vacuum reaction time 1/4~1/3) at synthesis under normal pressure, forward again vacuum reaction to, under the vacuum condition that is 10~1000pa in absolute pressure, carry out.The consumption of step (2) reaction times and enzyme is relevant, and conventionally, the consumption of immobilized enzyme is 0.2~5% of substrate weight, and the reaction times is 3~24 hours.Passed through step (2) reaction, the triglyceride level major part in reactive system is converted into partial glyceride, and in partial glyceride, the weight ratio of mono-glycerides and triglyceride is left and right, 1:4~8 conventionally.
Step (3) is separation phase, step (2) reaction product mainly comprises triglyceride level, triglyceride, mono-glycerides, fatty-acid ethyl ester, glycerine, conventionally triglyceride and mono-glycerides are as object product, and separated object is to obtain relatively pure triglyceride and mono-glycerides.Conventional separation method is molecular distillation, and the feature of these class methods is under vacuum condition, can significantly reduce separated needed temperature.Molecular distillation and short-path distillation separation method are the conventional separation unit operation of a class, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, and distillation temperature is 80~120 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 150~180 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 190~220 ℃.In the present invention, triglyceride and triglyceride level are to mix and exist as high temperature cut, in the present invention, containing the product of triglyceride, are the mixture of triglyceride and triglyceride level.
In the present invention, owing to there is no the participation of water, the free fatty acid content in each stage enzyme reaction thing is very low, and in the mono-glycerides product that separation obtains through step (3), main component is mono-glycerides, and free fatty acid content is lower than 1.5%; The high temperature cut part of step (3), free fatty acid content is lower than 0.5%.Because step (3) high temperature cut is as target product, for further improving quality standard, still need further refining treatment.In the present invention, step (3) high temperature cut need to be further through decolouring and deodorization processing, and Zhe Liang workshop section is edible oil processing common processes.The discoloring agent of the general employing 1.5% of decolouring is processed, and discoloring agent is the mixture of atlapulgite and gac 20:1.Deodorization is to adopt wet distillation method to remove the bad flavor of fat residue.
This research team carries out the glycerolysis reaction of grease for a long time, research discovery, adopting triglyceride level and glycerine is that substrate reacts, while there is free glycerol in reaction system, free glycerol is understood preferentially and zymin is wrapped up effect in conjunction with forming, and causes the apparent inactivation of enzyme.For addressing this problem, glycerine must adopt carrier to carry out preadsorption processing.Further research is found, fatty-acid ethyl ester is made an addition in the reaction system of triglyceride level and free glycerol, enzyme reactor adopts fixed-bed reactor, reaction substrate mixture is forced constantly to cycle through fixed bed enzyme reactor, now, although do not adopt the glycerine of ADSORPTION STATE, reaction still can be carried out smoothly.Show, the parcel effect of glycerine to zymin alleviated in the interpolation of fatty-acid ethyl ester, makes to take the glycerolysis reaction that free glycerol is substrate to be achieved.Because reaction process of the present invention does not have the participation of water, avoid the generation of free fatty acids, thereby can produce the reaction product of low acid value, obtain beyond thought effect, and then formed the present invention.
With existing technology, compare, the technology of the present invention has the following advantages:
Reaction system of the present invention does not need to add water, has solved product acid value problem, thereby can coproduction mono-glycerides; Because glycerine does not need to do preadsorption, do not process, simplified production operation.
Accompanying drawing explanation
Fig. 1 is the device of enzyme process glycerolysis reaction of the present invention, raw material storage tank 1, pump 2, packed bed enzyme reactor 3.
Embodiment
Below in conjunction with embodiment, the invention will be further described, and in embodiment, all % content specifies, is all weight percentage.
Embodiment 1
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
Step (1), 5kg soybean oil and 0.3kg dehydrated alcohol mix, and add immobilized lipase Novozyme 435(Denmark Novozymes company to provide) 2g, 35 ℃ of stirring reactions 3 hours, surveying fatty-acid ethyl ester production rate is 20.6%, and vacuum removal residual ethanol, obtains alcoholysis product.
Step (2), alcoholysis product 3kg packs in raw material storage tank 1 as shown in Figure 1, add 0.3kg glycerine, material cycles through the packed bed enzyme reactor 3 of immobilized lipase under pump 2 effects, and temperature of reaction is 60 ℃, adopt immobilized lipase Novozyme 435, the amount of fill of enzyme is 10g, reacts 1 hour under normal pressure, then will react absolute pressure and control as 10pa, react again discharging after 3 hours, analyze it and form in Table 1.
Step (3), reaction product is separated through molecular distillation, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, distillation temperature is 110 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 175 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 210 ℃.High temperature cut and low temperature fraction analytical results table 1.
Embodiment 2
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
Step (1), 5kg soybean oil and 0.3kg dehydrated alcohol mix, and add immobilized lipase Novozyme 435(Denmark Novozymes company to provide) 2g, 35 ℃ of stirring reactions 0.5 hour, surveying ethyl ester production rate is 9.8%, and vacuum removal residual ethanol, obtains alcoholysis product.
Step (2), alcoholysis product 3kg packs in raw material storage tank as shown in Figure 1, add 0.15kg glycerine, material cycles through the packed bed enzyme reactor of immobilized lipase under pumping action, and temperature of reaction is 60 ℃, adopt immobilized lipase Novozyme 435, the amount of fill of enzyme is 10g, first under normal pressure, reacts 6 hours, then reacts absolute pressure and controls as 100pa, react discharging after 24 hours, analyze it and form in Table 1.
Step (3), reaction product is separated through molecular distillation, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, distillation temperature is 110 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 175 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 210 ℃.High temperature cut and low temperature fraction analytical results table 1.
Embodiment 3
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
Step (1), 5kg soybean oil and 0.3kg dehydrated alcohol mix, and add immobilized lipase Novozyme 435(Denmark Novozymes company to provide) 2g, 35 ℃ of stirring reactions 5 hours, surveying ethyl ester production rate is 31.2%, and vacuum removal residual ethanol, obtains alcoholysis product.
Step (2), alcoholysis product 3kg packs in raw material storage tank as shown in Figure 1, add 0.45kg glycerine, material cycles through the packed bed enzyme reactor of immobilized lipase under pumping action, and temperature of reaction is 60 ℃, adopt immobilized lipase Novozyme 435, the amount of fill of enzyme is 10g, first under normal pressure, reacts 3 hours, then reacts absolute pressure and controls as 200pa, react discharging after 12 hours, analyze it and form in Table 1.
Step (3), reaction product is separated through molecular distillation, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, distillation temperature is 110 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 175 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 210 ℃.High temperature cut and low temperature fraction analytical results table 1.
Embodiment 4
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
Step (1), 5kg soybean oil and 0.3kg dehydrated alcohol mix, and add immobilized lipase Novozyme 435(Denmark Novozymes company to provide) 2g, 30 ℃ of stirring reactions 3 hours, surveying ethyl ester production rate is 18.3%, and vacuum removal residual ethanol, obtains alcoholysis product.
Step (2), alcoholysis product 3kg packs in raw material storage tank as shown in Figure 1, add 0.2kg glycerine, material cycles through the packed bed enzyme reactor of immobilized lipase under pumping action, and temperature of reaction is 60 ℃, adopt immobilized lipase Novozyme 435, the amount of fill of enzyme is 10g, first under normal pressure, reacts 2 hours, then reacts absolute pressure and controls as 1000pa, react discharging after 8 hours, analyze it and form in Table 1.
Step (3), reaction product is separated through molecular distillation, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, distillation temperature is 110 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 175 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 210 ℃.High temperature cut and low temperature fraction analytical results table 1.
Embodiment 5
A triglyceride Production by Enzymes method for the unsaturated mono-glycerides of coproduction, comprises the following steps:
Step (1), 5kg soybean oil and 0.3kg dehydrated alcohol mix, and add immobilized lipase Novozyme 435(Denmark Novozymes company to provide) 2g, 35 ℃ of stirring reactions 3 hours, surveying ethyl ester production rate is 22.3%, and vacuum removal residual ethanol, obtains alcoholysis product.
Step (2), alcoholysis product 3kg packs in raw material storage tank as shown in Figure 1, add 0.3kg glycerine, material cycles through the packed bed enzyme reactor of immobilized lipase under pumping action, and temperature of reaction is 60 ℃, adopt immobilized lipase Novozyme 435, the amount of fill of enzyme is 10g, first under normal pressure, reacts 3 hours, then reacts absolute pressure and controls as 100pa, react discharging after 12 hours, analyze it and form in Table 1.
Step (3), reaction product is separated through molecular distillation, and the present invention distills flow process can adopt three-stage distillation, the degassed and de-glycerine of the first step, distillation temperature is 110 ℃; The second stage removes fatty-acid ethyl ester, and distillation temperature is 175 ℃; Third stage separation obtains take triglyceride and triglyceride level is main high temperature cut, obtains take mono-glycerides as main low temperature fraction simultaneously, and distillation temperature is 210 ℃.High temperature cut and low temperature fraction analytical results table 1.
Comparative example
3kg soybean oil raw material packs in raw material storage tank as shown in Figure 1, add 0.15kg glycerine, material cycles through the packed bed enzyme reactor of immobilized lipase under pumping action, temperature of reaction is 60 ℃, adopts immobilized lipase Novozyme 435, and the amount of fill of enzyme is 10g, first under normal pressure, react 3 hours, then react absolute pressure and control as 100pa, react discharging after 12 hours, glycerine hydrolysis products analytical results is in Table 1.
Table 1
From table 1 result, comparative example directly adopts soybean oil to carry out enzyme process glycerine solution, and glycerolysis reaction does not almost occur.Embodiment 1 to embodiment 5 is first ethanolysis glycerine solution again, and reaction can be carried out smoothly, can obtain the grease that comprises triglyceride, obtains the mono-glycerides product of low acid value simultaneously.