CN108239578A - A kind of method for improving grease hydrolysis rate - Google Patents
A kind of method for improving grease hydrolysis rate Download PDFInfo
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- CN108239578A CN108239578A CN201611206801.9A CN201611206801A CN108239578A CN 108239578 A CN108239578 A CN 108239578A CN 201611206801 A CN201611206801 A CN 201611206801A CN 108239578 A CN108239578 A CN 108239578A
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- lipase
- oil
- grease
- thermomyces
- rhizomucor
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/02—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils
- C11C1/04—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by hydrolysis
- C11C1/045—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by hydrolysis using enzymes or microorganisms, living or dead
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- Life Sciences & Earth Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of methods for improving grease hydrolysis rate.Specifically, the present invention provides a kind of hydrolysate oil or the method for improving grease hydrolysis rate, and the method includes using the step of lipase of (a) from thermophilic fungal category (Thermomyces sp.) or the lipase from root Mucor (Rhizomucor sp.) are with (b) partial glyceride lipase progress grease hydrolysis.The present invention also provides a kind of lipase compositions, the lipase belonged to containing (a) from thermophilic fungal or the lipase from root Mucor；(b) partial glyceride lipase；Wherein, it is not 1 that the lipase of thermophilic fungal category is originated from the composition with the weight ratio of the partial glyceride lipase.Method and composition using the present invention can improve the percent hydrolysis of grease, so as to greatly improve production efficiency in the case where saving production cost.
Description
Technical field
The invention belongs to grease processing technique field, specially a kind of method for improving grease hydrolysis rate
Background technology
Ricinoleic acid is common raw material in oiling industry, is mainly used to prepare decanedioic acid and 12- hydroxy stearic acids.Castor-oil plant
Oleic acid is the hydrolysate of castor oil, and in castor oil, the content of fatty acid of ricinoleic acid accounts for more than 90%, far above other oil kind
Content.
The main preparation methods of ricinoleic acid are high temperature and high pressure method and enzymatic hydrolysis at present.Although high temperature and high pressure method production effect
Rate is high, but this method condition is harsh, and hydroxyl and carboxylic in ricinoleic acid can be caused during being hydrolyzed applied to castor oil
Base occurs esterification and forms polyester, and so as to cause the color of product deeper, but also it is hard to reduce later stage preparation 12- hydroxyls
The yield of resin acid.
Compared to high temperature and high pressure method, enzymatic hydrolysis castor oil can obtain the product of high quality under mild conditions, simultaneously
The generation of waste water can be reduced, reduces the pollution to environment.But there is also problems with for enzymatic hydrolysis：It is universal in hydrolytic process
There is a situation where that hydrolysis efficiency is low, be unable to reach more than 90%；Even if percent hydrolysis can reach 90%, then enzyme concentration can compare
Greatly, it is of high cost, can not meet the needs of production.
For example, CN 200710118466.1 describes a kind of method of hydrolyzed castor oil, this method uses combined lipase
(Novozyme 435, Lipozyme RM IM, Lipozyme TL) is hydrolyzed, and for controlling reaction temperature at 35-60 DEG C, water is oily
Molar ratio is 1-50, and the reaction time, last yield was 90% or so in 10-30h.But what this method was added in hydrolytic process
Fatty enzyme amount is larger, of high cost, is not suitable for industrialized production.
CN 201080010073.9 describes a kind of method that ricinoleate ester is prepared by transesterification.Main technique walks
Rapid is that short chain alcohol is added in castor oil, and ricinoleate ester is generated by alcoholysis reaction under the action of lipase.
Yang Wei etc. (" free-fat enzymatic castor oil prepares ricinoleic acid ",《Biotechnological Advances》, 2014 (5):373-
378) principal element of castor oil hydrolytic process is catalyzed to influencing free-fat enzyme NS81006：Temperature, enzyme dosage, water consumption and
Mixing speed is studied and has been optimized, after oiling under conditions of 48h percent hydrolysis reach 94.8%.But during this method reaction
Between it is long, influence production efficiency in the actual production process.
(" the Enzymatic hydrolysis of castor oil such as Puthli M S:Process
Intensification studies ", Biochemical Engineering Journal, 2006,31 (1):It is 31-41) right
Each factor of enzymatic hydrolysis castor oil：The factors such as temperature, time, grease molar ratio, acetone additive amount, enzyme additive amount carry out
It investigates.Percent hydrolysis can not meet normal production requirement all than relatively low in this article report.
Therefore, this field still needs a kind of method of enzymatic hydrolysis grease, and production cost can be being saved using this method
In the case of improve grease percent hydrolysis, so as to greatly improve production efficiency.
Invention content
It is compounded herein using two kinds of lipase, gives full play to two kinds of lipase respectively characteristic, it is seldom in enzyme additive amount
In the case of can reach very high percent hydrolysis in a short time, improve production efficiency, save production cost.
Therefore, first aspect present invention provides a kind of grease hydrolysis method, the method includes using lipase (a), i.e.,
Lipase from thermophilic fungal category (Thermomyces sp.) or the fat from root Mucor (Rhizomucor sp.)
Enzyme, and lipase (b), i.e. partial glyceride lipase, the step of carrying out grease hydrolysis.
Second aspect of the present invention provides a kind of method for improving grease hydrolysis rate, and the method includes using lipase (a)
The step of carrying out grease hydrolysis with lipase (b), fat of the lipase (a) from thermophilic fungal category (Thermomyces sp.)
Fat enzyme or the lipase from root Mucor (Rhizomucor sp.), the lipase (b) are partial glyceride lipases.
In one or more embodiments, the lipase from thermophilic fungal category (Thermomyces sp.) is
Lipase from Thermomyces lanuginosus (Thermomyces languginosus).
In one or more embodiments, the lipase from thermophilic fungal category (Thermomyces sp.) is
Lipozyme TL。
In one or more embodiments, the lipase from root Mucor (Rhizomucor sp.) is source
From the lipase of Rhizomucor miehei (Rhizomucor miehei).
In one or more embodiments, the lipase from root Mucor (Rhizomucor sp.) is
Lipozyme RM IM。
In one or more embodiments, the partial glyceride lipase is derived from Penicillium (Penicilliumsp.)
Lipase.
In one or more embodiments, the lipase from Penicillium (Penicillium sp.) is derived from
The lipase of penicillium cammenberti (Penicillium camemberti).
In one or more embodiments, the lipase from Penicillium (Penicillium sp.) is fat
Enzyme G50, i.e. commercial enzyme Lipase G " Amano " 50.
In one or more embodiments, the additive amount of the lipase (a) is the 0.2~5% of grease weight, example
Such as 0.2~4%, 0.2~3%, 0.2~2%, 0.2~1%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.5~
5%th, 0.5~4%, 0.5~3%, 0.5~2.5%, 0.5~2%, 0.5~1.5%, 0.5~1%, 0.8~5%, 0.8~
4%th, 0.8~3%, 1~5%, 1~4% or 1~3%.
In one or more embodiments, the additive amount of the lipase (b) for grease weight 0.15~
2.0%, such as 0.15~1.5%, 0.15~1.2%, 0.15~1.0%, 0.15~0.8%, 0.15~0.6%, 0.15~
0.5%th, 0.15~0.3%, 0.2~1.8%, 0.2~1.5%, 0.2~1.0%, 0.2~0.8%, 0.2~0.6%, 0.2
~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~1.2%, 0.3~1.0%, 0.5~2.0%, 0.5~1.8%, 0.5
~1.5%, 0.5~1.2%, 0.5~1.0%.
In one or more embodiments, hydrolysis carries out in presence of water.
In one or more embodiments, the mass ratio of grease and water is 1：0.2~5, such as 1：0.2~4,1：0.2
~3,1：0.3~5,1：0.3~4,1：0.3~3,1：0.4~5,1：0.4~4,1：0.4~3 etc..
It in one or more embodiments, hydrolyzes and is carried out at a temperature of 35~55 DEG C, such as in 40~50 DEG C of temperature
Degree is lower to carry out.
In one or more embodiments, the grease is vegetable oil or animal oil.
In one or more embodiments, the grease is polished fat.
In one or more embodiments, the grease is selected from：Castor oil, Rice oil, sunflower oil, palm oil, palm fibre
Palmitic acid benevolence oil, peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, Purple Perilla Seed Oil, tea-seed oil, palm fruit oil, coconut oil, olive
Oil, oleum theobromatis, tallowseed oil, almond oil, apricot kernel oil, bancoul nuts oil, rubber seed oil, maize germ, wheat germ oil, sesame
Seed oil, linseed oil, oenothera seed oil, hazelnut oil, nut oil, grape seed oil, borage seed oil, Seabuckthorm Seed Oil, tomato seed oil,
Pumpkin seed oil, macadimia nut oil, cocoa butter, algae oil, tallow, lard, sheep oil, chicken fat, fish oil, seal oil, whale oil, dolphin oil
With the arbitrary mixing of the grease of one or more of oyster sauce.
In one or more embodiments, the grease is castor oil.
In one or more embodiments, the method includes using to be originated from thermophilic fungal category (Thermomyces
Sp. the step of lipase) carries out grease hydrolysis with partial glyceride lipase.
In one or more embodiments, the method includes using to be originated from root Mucor (Rhizomucorsp.)
Lipase and partial glyceride lipase the step of carrying out grease hydrolysis；Preferably, it in one or more embodiments, is originated from
The dosage of the lipase of root Mucor (Rhizomucor sp.) is the 0.8~5% of grease weight, such as 0.8~4%, 0.8
~3%, 1~5%, 1~4% or 1~3%；Preferably, in one or more embodiments, the use of partial glyceride lipase
Measure 0.15~2% for grease weight, such as 0.15~1%, 0.3~2%, 0.3~1.5%, 0.3~1.2% or 0.3~
1.0%.
Third aspect present invention provides partial glyceride lipase or its combination with following lipase in grease hydrolysis or carries
Application in high grease hydrolysis rate：
Lipase (a), i.e., lipase from thermophilic fungal category (Thermomyces sp.) or from root Mucor
The lipase of (Rhizomucor sp.)；With
Lipase (b), i.e. partial glyceride lipase.
Fourth aspect present invention provides a kind of lipase compositions, and the composition contains：
Lipase (a), i.e., lipase from thermophilic fungal category (Thermomyces sp.) or from root Mucor
The lipase of (Rhizomucor sp.)；With
Lipase (b), i.e. partial glyceride lipase；
Wherein, the lipase in the composition from thermophilic fungal category (Thermomyces sp.) and the inclined glycerine
The weight ratio of ester lipase is not 1.
In one or more embodiments, the composition contains lipase (a) and lipase (b)：
Lipase (a) is derived from the lipase of root Mucor (Rhizomucor sp.)；With
Lipase (b) is partial glyceride lipase.
In one or more embodiments, the lipase from thermophilic fungal category (Thermomyces sp.) is
Lipase from Thermomyces lanuginosus (Thermomyces languginosus).
In one or more embodiments, the lipase from thermophilic fungal category (Thermomyces sp.) is
Lipozyme TL。
In one or more embodiments, the lipase from root Mucor (Rhizomucor sp.) is source
From the lipase of Rhizomucor miehei (Rhizomucor miehei).
In one or more embodiments, the lipase from root Mucor (Rhizomucor sp.) is
Lipozyme RM IM。
In one or more embodiments, the partial glyceride lipase is derived from Penicillium (Penicilliumsp.)
Lipase.
In one or more embodiments, the lipase from Penicillium (Penicillium sp.) is derived from
The lipase of penicillium cammenberti (Penicillium camemberti).
In one or more embodiments, the lipase from Penicillium (Penicillium sp.) is fat
Enzyme G50 (Lipase G " Amano " 50).
Specific embodiment
Lipase
Herein, " partial glyceride lipase " refers to do not have catalyzing hydrolysis vigor to triglycerides, preferentially to diglyceride
And/or monoglyceride has the lipase of catalyzing hydrolysis vigor.Partial glyceride lipase mainly has two classes：Monoglyceride-glycerine
Diester lipase and monoglyceride lipase.Present invention preferably uses have catalyzing hydrolysis to monoglyceride and/or diglyceride
The partial glyceride lipase of vigor.Partial glyceride lipase well known in the art can be used, for example, can be used from aspergillus
(Aspergillus), the partial glyceride lipase of Penicillium (Penicillium) and Marxist philosophy lesson (Malassezia).
In certain embodiments, it is suitable for the invention partial glyceride lipase and includes but not limited to such as CN
Partial glyceride lipase disclosed in 201410182887.0, commercially available partial glyceride lipase such as lipase G50 (Lipase
And lipase GH1 (Huang J, Yang Z, Guan F etc., A novel mono-and G " Amano " 50)
diacylglycerol lipase highly expressed in Pichia pastoris,and its application
For food emulsifier preparation, Process Biochemistry, 2013,48 (12):1899-1904)、
Lipase SMG1 (Xu T, Lu L, Hou S etc., Crystal structure of a mono-and diacylglycerol
lipase from Malassezia globosa,reveals a novel lid conformation and insights
Into the substrate specificity, Journal of Structural Biology, 2012,178 (3):363-
And lipase rePcMdl (Tan Z B, Li J F, Li X T etc., A unique mono-and diacylglycerol 9)
lipase from Penicillium cyclopium:heterologous expression,biochemical
Characterization and molecular basis for its substrate selectivity, Plos One,
2014,9 (7):E102040) etc..
During in the method or purposes of the present invention, the dosage of partial glyceride lipase be usually grease weight 0.12~
2.0%, such as 0.15~2.0%, 0.15~1.5%, 0.15~1.2%, 0.15~1.0%, 0.15~0.8%, 0.15~
0.6%th, 0.15~0.5%, 0.15~0.3%, 0.2~1.8%, 0.2~1.5%, 0.2~1.0%, 0.2~0.8%, 0.2
~0.6%, 0.2~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~1.2%, 0.3~1.0%, 0.5~2.0%, 0.5
~1.8%, 0.5~1.5%, 0.5~1.2%, 0.5~1.0% etc..
In certain embodiments, the present invention uses the partial glyceride lipase from Penicillium (Penicillium sp.)
Enzyme, especially from the partial glyceride lipase of penicillium cammenberti (Penicillium camemberti).For example, it comes from
The partial glyceride lipase of Penicillium (Penicillium sp.) can be lipase G50, i.e. Lipase G " Amano " 50.
In the present invention, thermophilic fungal category can be derived from another lipase of partial glyceride lipase compounding
The lipase of (Thermomyces sp.) or the lipase from root Mucor (Rhizomucor sp.).In certain implementations
In scheme, the lipase from thermophilic fungal category (Thermomyces sp.) is derived from Thermomyces lanuginosus
The lipase of (Thermomyces languginosus), for example, it is described from thermophilic fungal category (Thermomyces sp.)
Lipase is Lipozyme TL IM or derives from thermophilic fungal category with similar zymologic property with Lipozyme TL IM
The lipase of (Thermomyces sp.).
In certain embodiments, the lipase from root Mucor (Rhizomucor sp.) can be derived from meter He
The lipase of root Mucor (Rhizomucor miehei), for example, the fat from root Mucor (Rhizomucor sp.)
Fat enzyme is Lipozyme RM IM or derives from root Mucor with similar zymologic property with Lipozyme RM IM
Lipase.
During in the method or purposes of the present invention, lipase from thermophilic fungal category (Thermomyces sp.) or
The dosage of lipase from root Mucor (Rhizomucor sp.) can be the 0.15~5% of grease weight, such as 0.2
~5%, 0.2~4%, 0.2~3%, 0.2~2%, 0.2~1%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%,
0.5~5%, 0.5~4%, 0.5~3%, 0.5~2.5%, 0.5~2%, 0.5~1.5%, 0.5~1%, 0.8~5%,
0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3% etc..
In certain embodiments, the present invention is used using the lipase from root Mucor (Rhizomucor sp.)
Amount can be the 0.8~5% of grease weight, such as 0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3%.
In certain embodiments, the present invention is using partial glyceride lipase and from thermophilic fungal category (Thermomyces
Sp. the combination of lipase).In these embodiments, the dosage of partial glyceride lipase can be the 0.15 of grease weight
~2.0%, such as 0.15~1.5%, 0.15~1.2%, 0.15~1.0%, 0.15~0.8%, 0.15~0.6%, 0.15
~0.5%, 0.15~0.3%, 0.2~1.8%, 0.2~1.5%, 0.2~1.0%, 0.2~0.8%, 0.2~0.6%,
0.2~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~1.2%, 0.3~1.0%, 0.5~2.0%, 0.5~1.8%,
0.5~1.5%, 0.5~1.2% or 0.5~1.0%；The use of lipase from thermophilic fungal category (Thermomyces sp.)
Amount can be grease weight 0.2~5%, such as 0.2~4%, 0.2~3%, 0.2~2%, 0.2~1%, 0.2~
0.8%th, 0.2~0.6%, 0.2~0.5%, 0.5~5%, 0.5~4%, 0.5~3%, 0.5~2.5%, 0.5~2%,
0.5~1.5%, 0.5~1%, 0.8~5%, 0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3%.
The arbitrary combination of above-mentioned two kinds of listed lipase amount ranges can be used to implement the present invention.For example, partially
The dosage of glyceride fat enzyme can be the 0.15~1.5% of grease weight, such as 0.15~1.0%, the matched source used
From the dosage of the lipase of thermophilic fungal category (Thermomyces sp.) can be grease weight 0.2~5%, 0.2~4%,
0.2~3%, 0.2~2%, 0.2~1%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.5~5%, 0.5~
4%th, 0.5~3%, 0.5~2.5%, 0.5~2%, 0.5~1.5% or 0.5~1%；Similarly, from thermophilic fungal category
The dosage of the lipase of (Thermomyces sp.) can be the 0.2~5% of grease weight, such as 0.2~2.0%, and match therewith
It can be the 0.15~2.0% of grease weight, such as 0.15~1.5%, 0.15 to close the dosage of partial glyceride lipase used
~1.2%, 0.15~1.0%, 0.15~0.8%, 0.15~0.6%, 0.15~0.5%, 0.15~0.3%, 0.2~
1.8%th, 0.2~1.5%, 0.2~1.0%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.3~2.0%, 0.3~
1.5%th, 0.3~1.2%, 0.3~1.0%, 0.5~2.0%, 0.5~1.8%, 0.5~1.5%, 0.5~1.2% or 0.5
~1.0%.
In certain embodiments, the dosage of partial glyceride lipase can be the 0.15~1% of grease weight, such as 0.15
~0.6%, 0.15~0.5% or 0.15~0.3%, it is matched use be originated from thermophilic fungal category (Thermomyces
Sp. the dosage of lipase) can be grease weight 0.2~2%, such as 0.2~1%, 0.2~0.8%, 0.2~0.6%,
0.2~0.5%, 0.5~2%, 0.5~1.5% or 0.5~1%.
In certain embodiments, the present invention is using partial glyceride lipase and from root Mucor (Rhizomucor
Sp. the combination of lipase).In these embodiments it is preferred that the dosage of partial glyceride lipase is grease weight
0.15~2%, such as 0.15~1%, 0.3~2%, 0.3~1.5%, 0.3~1.2% or 0.3~1.0%；It is matched to make
The dosage for being originated from the lipase of root Mucor (Rhizomucor sp.) can be the 0.8~5% of grease weight, such as
0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3%.
In general, the percentage that the total weight for all lipase being added in grease accounts for grease weight is no more than 7.0%, example
Such as between 0.35~7.0%, such as 0.35~6.0%, 0.35~5.0%, 0.35~4.0%, 0.35~3.0%, 0.35~
2.0%th, 0.35~1.5%, 0.35~1.0%, 0.35~0.8%, 0.35~0.5%, 0.4~7.0%, 0.4~6.0%,
0.4~5.0%, 0.4~4.0%, 0.5~6.0%, 0.5~5.0%, 0.8~6.0%, 0.8~5.0%, 1.0~6.0%
Deng in the range of.
In certain embodiments, the fat from thermophilic fungal category (Thermomyces sp.) being added in grease
The weight of enzyme or lipase from root Mucor (Rhizomucor sp.) is higher than the partial glyceride being added in grease
The weight of lipase.For example, in these embodiments, it is added in grease and is originated from thermophilic fungal category (Thermomyces
Sp. the weight of lipase) or the lipase from root Mucor (Rhizomucor sp.) be added to it is inclined in grease
At least 1.1 times of the weight of glyceride fat enzyme, such as 1.1~33.3 times, such as 1.1~30 times, 1.1~25 times, 1.1~20
Times, 1.1~15 times, 1.1~10 times etc..
In certain embodiments, the lipase of thermophilic fungal category (Thermomyces sp.) and described herein will be originated from
Partial glyceride lipase be added in grease, the former additive amount is at least 1.2 times, for example, at least 1.3 of the latter's additive amount
Times, such as 1.2~33.3 times, such as 1.1~30 times, 1.1~25 times, 1.1~20 times, 1.1~15 times, 1.1~10 times, 1.2
~5 times, 1.2~3 times etc..In other embodiments, the additive amount of partial glyceride lipase is derived from thermophilic fungal category
At least 1.1 times of the lipase additive amount of (Thermomyces sp.), for example, at least 1.2 times, such as 1.1~10 times, 1.1~
8 times, 1.1~5 times, 1.1~3 times, 1.2~8 times etc..
In other embodiments, the lipase of root Mucor (Rhizomucor sp.) and as described herein will be originated from
Partial glyceride lipase is added in grease, the former additive amount is at least 1.5 times of the latter's additive amount, for example, at least 2 times, example
Such as 2~33.3 times, such as 1.5~30 times, 1.5~25 times, 1.5~20 times, 1.5~15 times, 1.5~10 times, 3~10 times etc..
It should be understood that in these embodiments, the lipase added while the multiple requirement is met,
Dosage should be also fallen within previously described amount ranges herein.
Lipase for the present invention can be used with various forms well known in the art.For example, lipase used can be with
For one or more mixing such as liquid enzymes, powdered enzyme, solid enzyme, immobilised enzymes.
Lipase compositions
The present invention also provides a kind of lipase compositions, and the lipase contained by the composition is (a) from thermophilic fungal category
The lipase of (Thermomyces sp.) or lipase and (b) inclined glycerine from root Mucor (Rhizomucor sp.)
Ester lipase.
In general, the percentage that the lipase contained by the lipase compositions being added in grease accounts for grease weight is no more than
7.0%, such as between 0.35~7.0%, such as 0.35~6.0%, 0.35~5.0%, 0.35~4.0%, 0.35~
3.0%th, 0.35~2.0%, 0.35~1.5%, 0.35~1.0%, 0.35~0.8%, 0.35~0.5%, 0.4~7.0%,
0.4~6.0%, 0.4~5.0%, 0.4~4.0%, 0.5~6.0%, 0.5~5.0%, 0.8~6.0%, 0.8~5.0%,
In the range of 1.0~6.0% grades.
In lipase compositions of the present invention the content of lipase (a) should be sufficient to make by the composition with contained lipase
Account for the lipase (a) when the amount of the percentage of grease weight no more than 7% is added in grease account for grease weight 0.2~
5%, for example, 0.2~4%, 0.2~3%, 0.2~2%, 0.2~1%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%,
0.5~5%, 0.5~4%, 0.5~3%, 0.5~2.5%, 0.5~2%, 0.5~1.5%, 0.5~1%, 0.8~5%,
0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3% etc..
The content of lipase (b) in lipase compositions of the present invention, i.e. partial glyceride lipase should be sufficient to make should
Composition accounts for the partial glyceride lipase when amount of the percentage of grease weight no more than 7% is added in grease with contained lipase
Enzyme accounts for the 0.15~2.0% of grease weight, such as 0.15~1.5%, 0.15~1.2%, 0.15~1.0%, 0.15~
0.8%th, 0.15~0.6%, 0.15~0.5%, 0.15~0.3%, 0.2~1.8%, 0.2~1.5%, 0.2~1.0%,
0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~1.2%, 0.3~1.0%,
0.5~2.0%, 0.5~1.8%, 0.5~1.5%, 0.5~1.2% or 0.5~1.0% etc..
In certain embodiments, the lipase (a) described in lipase compositions is derived from thermophilic fungal category
The lipase of (Thermomyces sp.), content should be sufficient to make is accounting for oil by the lipase compositions with contained lipase
Amount of the percentage of fat weight no more than 7% is originated from the fat of thermophilic fungal category (Thermomyces sp.) when being added in grease
Fat enzyme accounts for the 0.2~5% of grease weight, for example, 0.2~4%, 0.2~3%, 0.2~2%, 0.2~1%, 0.2~0.8%,
0.2~0.6%, 0.2~0.5%, 0.5~5%, 0.5~4%, 0.5~3%, 0.5~2.5%, 0.5~2%, 0.5~
1.5%th, 0.5~1%, 0.8~5%, 0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3% etc.；In lipase (b)
The content of the partial glyceride lipase in the composition should be sufficient to make by the lipase compositions with contained lipase
It accounts for partial glyceride lipase described when the amount of the percentage of grease weight no more than 7% is added in grease and accounts for grease weight
0.15~2.0%, for example, 0.15~1.5%, 0.15~1.2%, 0.15~1.0%, 0.15~0.8%, 0.15~0.6%,
0.15~0.5%, 0.15~0.3%, 0.2~1.8%, 0.2~1.5%, 0.2~1.0%, 0.2~0.8%, 0.2~
0.6%th, 0.2~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~1.2%, 0.3~1.0%, 0.5~2.0%, 0.5~
1.8%th, 0.5~1.5%, 0.5~1.2% or 0.5~1.0%
In other embodiments, the lipase (a) described in lipase compositions of the present invention is from root Mucor
The lipase of Pseudomonas (Rhizomucor sp.), the content of the lipase in the composition should be sufficient to make by the lipase group
Close object with contained lipase account for when the amount of the percentage of grease weight no more than 7% is added in grease described in be originated from root mucor
The lipase for belonging to (Rhizomucor sp.) accounts for the 0.8~5% of grease weight, such as 0.8~4%, 0.8~3%, 1~5%, 1
~4% or 1~3%；Content of the partial glyceride lipase in the lipase compositions described in lipase (b) should be enough to make
When the lipase compositions being accounted for the amount of the percentage of grease weight no more than 7% with contained lipase being added in grease
The partial glyceride lipase accounts for the 0.15~2% of grease weight, such as 0.15~1%, 0.3~2%, 0.3~1.5%,
0.3~1.2% or 0.3~1.0%.
In certain embodiments, thermophilic fungal category (Thermomyces sp.) is originated from lipase compositions of the present invention
The weight ratio of lipase and partial glyceride lipase as described herein be not 1.In certain embodiments, lipase of the present invention
The lipase of thermophilic fungal category (Thermomyces sp.) is originated from composition or from root Mucor (Rhizomucor
Sp. the weight of lipase) is higher than the weight of partial glyceride lipase.For example, in these embodiments, from thermophilic fungal
Belong to (Thermomyces sp.) lipase or lipase from root Mucor (Rhizomucor sp.) with it is described herein
Partial glyceride lipase weight ratio 1.1~33.3：1, such as 1.1~30：1,1.1~25：1,1.1~20：1,1.1~
15：1,1.1~10：1 etc..
In certain embodiments, lipase compositions of the present invention contain from thermophilic fungal category (Thermomyces
Sp. lipase) and partial glyceride lipase as described herein, the former weight are at least 1.2 times, such as extremely of the latter's weight
It is 1.3 times few, such as 1.2~33.3 times, such as 1.1~30 times, 1.1~25 times, 1.1~20 times, 1.1~15 times, 1.1~10 times
Deng.In other embodiments, the weight of partial glyceride lipase is derived from the fat of thermophilic fungal category (Thermomyces sp.)
At least 1.1 times of fat enzyme weight, for example, at least 1.2 times, such as 1.1~10 times, 1.1~8 times, 1.1~5 times, 1.1~3 times,
1.2~8 times etc..
In other embodiments, lipase compositions of the present invention contain from root Mucor (Rhizomucorsp.)
Lipase and partial glyceride lipase as described herein, the former weight be at least 1.5 times, for example, at least 2 of the latter's weight
Times, such as 2~33.3 times, such as 1.5~30 times, 1.5~25 times, 1.5~20 times, 1.5~15 times, 1.5~10 times etc..
It should be understood that in these embodiments, the weight of the lipase in lipase compositions is meeting described times
While number requires, content should be also fallen within previously described content range herein.
The present invention lipase compositions can be various lipase compositions forms well known in the art, including but it is unlimited
In the form of liquid composition, solid composite (such as powder composition) or immobilised enzymes.
Method for hydrolysis
The lipase or combination object of the present invention can be used for hydrolysate oil or the percent hydrolysis available for improving grease.
It can be various greases well known in the art suitable for the grease of the method for the present invention and purposes, including vegetable oil or move
Object oil.Preferably, it is liquid under the hydrolysis temperature of the present invention for the grease of the present invention.Grease can also be refined oil
Fat.For example, grease is suitable for the invention may be selected from：Castor oil, Rice oil, sunflower oil, palm oil, palm-kernel oil,
Peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, Purple Perilla Seed Oil, tea-seed oil, palm fruit oil, coconut oil, olive oil, cocoa bean
Oil, tallowseed oil, almond oil, apricot kernel oil, bancoul nuts oil, rubber seed oil, maize germ, wheat germ oil, sesame seed oil, flax
Seed oil, oenothera seed oil, hazelnut oil, nut oil, grape seed oil, borage seed oil, Seabuckthorm Seed Oil, tomato seed oil, pumpkin seed oil,
In macadimia nut oil, cocoa butter, algae oil, tallow, lard, sheep oil, chicken fat, fish oil, seal oil, whale oil, dolphin oil and oyster sauce
One or more kinds of greases arbitrary mixing.In one or more embodiments, the grease is castor oil.
During hydrolysis, first according to situations such as specifically used enzyme, grease, suitable water is added in grease.In general, grease
Mass ratio with water is 1：0.2~5, such as 1：0.2~4,1：0.2~3,1：0.3~5,1：0.3~4,1：0.3~3,1：0.4
~5,1：0.4~4,1：0.4~3,1：0.6~3,1：0.6~2 etc..To obtain best hydrolysis effect, those skilled in the art
Grease and water can be determined and adjust according to specifically used grease, the dosage of enzyme, the temperature of reaction, time etc. factor of reaction
Mass ratio.
Then lipase can be added in grease.It should be understood that above-mentioned lipase (a) and (b) can be respectively added to grease
In, can also mixture form addition or can by the present invention lipase compositions be added in grease.Lipase adds
Dosage is as mentioned before.After addition, it is uniformly mixed grease, lipase and water.For example, can high speed shear, make its be uniformly mixed.
If lipase is added in a manner of aqueous solution in grease, it is preferred that ensure water in fatty aqueous acid with it is described previously
The water additionally added total amount and grease mass ratio within above range.
Hydrolysis can usually carry out at a temperature of 35~55 DEG C, such as can be carried out at a temperature of 40~50 DEG C.Cause
This, after by grease, lipase and water mixing, carries out in the range of the temperature of gained mixture is gradually warming up to 35~55 DEG C
Reaction.In general, warming while stirring.Sustainable stirring in reaction process.
The time of hydrolysis is had no it is specifically limited, can be according to the amount of the grease being hydrolyzed, lipase used
The factors such as type and its dosage are adjusted.Alternatively, sampling analysis sample acid value and hydrolysis can be calculated in hydrolysis reaction
Rate, after expected percent hydrolysis is reached, you can stop reaction.
In addition, the dosage of each enzyme can be suitably adjusted according to fat type, and the dosage of water, the temperature of hydrolysis and time etc.,
To reach best percent hydrolysis.In certain embodiments, using the method for the present invention, at least 90% percent hydrolysis can be realized,
Preferably at least 92% percent hydrolysis, more preferably at least 95% percent hydrolysis, more preferably at least 97% percent hydrolysis, more preferably at least
98% percent hydrolysis.
Therefore, the method for hydrolysate oil of the present invention or the method for improving grease hydrolysis rate include the use of (a) from thermophilic true
The lipase of Pseudomonas (Thermomyces sp.) or lipase from root Mucor (Rhizomucor sp.) and (b) are inclined
Glyceride fat enzyme carries out the step of grease hydrolysis.In certain embodiments, the method includes described in mixing grease, addition
It is warming up to 35~55 DEG C after the mixture of lipase and water, and reacts 6 hours at such a temperature, 8 hours, 12 hours, 16 hours,
24 hours or longer time or these combinations of values period, so as to make grease hydrolysis.
In certain embodiments, the percent hydrolysis of grease can be increased to more than 90% by method using the present invention.At certain
In a little embodiments, the percent hydrolysis of grease can be increased to more than 92% by method using the present invention.Method using the present invention
The percent hydrolysis of grease can be increased to more than 95%.Method using the present invention the percent hydrolysis of grease can be increased to 97% with
On, it is preferably increased to more than 98%.
In certain embodiments, method using the present invention, it is 1 to control the mass ratio of grease and water：0.6~3 or 1：
0.6~2, the additive amount of the lipase from thermophilic fungal category (Thermomyces sp.) account for grease weight 0.2~
2%, the additive amount of the lipase (b) accounts for the 0.15~1% of grease weight, can be by the hydrolysis of grease after reacting appropriate time
Rate is increased to 95% or more than 98%.
In certain embodiments, method using the present invention, it is 1 to control the mass ratio of grease and water：0.6~3 or 1：
0.6~2, the additive amount of the lipase from root Mucor (Rhizomucor sp.) accounts for the 1~5% of grease weight,
The additive amount of the lipase (b) accounts for the 0.15~1% of grease weight, after reacting appropriate time, can carry the percent hydrolysis of grease
Height is to 95% or more than 98%.
Using
The present invention also provides application of the partial glyceride lipase in hydrolysate oil and preparing for hydrolysate oil
Application in lipase compositions.There is also provided (a) lipase or source from thermophilic fungal category (Thermomyces sp.)
From the lipase of root Mucor (Rhizomucor sp.) and application of (b) partial glyceride lipase in grease hydrolysis, with
And the application in the lipase compositions for hydrolysate oil are prepared.The thermophilic fungal category (Thermomyces sp.)
Lipase or lipase from root Mucor (Rhizomucor sp.) can be as mentioned before with partial glyceride lipase.
Hereafter the present invention will be illustrated in a manner of specific embodiment.It should be understood that these embodiments are only illustrative, and
The range being not intended to limit the invention.Lipase Novozyme 435 used, Lipozyme RM IM, fat in embodiment
Fat enzyme Lipozyme TL IM are letter (China) Investment Co., Ltd of Novi product, and lipase G50 is Lipase G " Amano "
50, it is Amano Enzyme Inc. (Amano Enzyme Inc.) product.Castor oil used is available from Feng Yijing in embodiment
(Lianyun Harbour) Co., Ltd is learned in refinement；Coconut oil is purchased from Southseas Specialty Fats Industrial (Shanghai) Co., Ltd.；Fish oil purchased from BASF (in
State) Co., Ltd, trade name is DHA fish oil；Purified soyabean oil is purchased from Shanghai Jia Li grain and oil Co., Ltd, other conventional chemicals
Product are purchased from traditional Chinese medicines chemical reagent Co., Ltd, to analyze pure grade.Percentage in embodiment is weight percentage.Lipase TL
It is to add as an aqueous solution, first Lipozyme TL IM is dissolved in the water to form the aqueous solution of normal concentration.Other fat
Enzyme adds in powder form.
In embodiment, percent hydrolysis is calculated by the following formula：
Wherein, AV0Refer to glyceride stock acid value, be 1.70mgKOH/g for castor oil；AVtRefer to certain time interval t samplings
Acid value；SV refers to glyceride stock saponification number, is 182.01mgKOH/g for castor oil.
Comparative example 1
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase TL (Lipozyme
TL IM account for the 5% of castor-oil plant weight of oil), then it is thoroughly mixed substrate in 15000rpm down cuts 1min, then in stirring shape
It is heated to 45 DEG C under state to start to react, stops heating after reacting 6h, samples the centrifugation 3min under 10000rpm and is detached, taken
Layer oil phase detection acid value is 116.69mgKOH/g, and it is 63.77% to calculate percent hydrolysis.
Comparative example 2
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.6g lipase G50 (Lipase
G " Amano " 50, accounts for the 2% of castor-oil plant weight of oil), substrate then is thoroughly mixed in 15000rpm down cuts 1min, is then being stirred
It mixes and 45 DEG C are heated under state start to react, stop heating after reacting 6h, sample and 3min is centrifuged under 10000rpm detached,
It is 53.06mgKOH/g to take upper oil phase detection acid value, and it is 28.46% to calculate percent hydrolysis.
Embodiment 1
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase TL and (account for castor oil
The 5% of weight), 0.6g lipase G50 (account for castor-oil plant weight of oil 2%), then make substrate complete in 15000rpm down cuts 1min
Full mixing, 45 DEG C are then heated under stirring and starts to react, and stop heating after reacting 6h, sample under 10000rpm from
Heart 3min is detached, and it is 177.25mgKOH/g to take upper oil phase detection acid value, and it is 97.36% to calculate percent hydrolysis.
Embodiment 2
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase TL and (account for castor oil
The 5% of weight), 0.45g lipase G50 (account for castor-oil plant weight of oil 1.5%), then make substrate in 15000rpm down cuts 1min
It is thoroughly mixed, 45 DEG C is then heated under stirring and starts to react, stop heating after reacting 8h, sample under 10000rpm
Centrifugation 3min is detached, and it is 173.61mgKOH/g to take upper oil phase detection acid value, and it is 95.34% to calculate percent hydrolysis.
Embodiment 3
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase TL and (account for castor oil
The 5% of weight), 0.3g lipase G50 (account for castor-oil plant weight of oil 1%), then make substrate complete in 15000rpm down cuts 1min
Full mixing, 45 DEG C are then heated under stirring and starts to react, and stop heating after reacting 8h, sample under 10000rpm from
Heart 3min is detached, and it is 169.95mgKOH/g to take upper oil phase detection acid value, and it is 93.31% to calculate percent hydrolysis.
Embodiment 4
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.3g lipase TL and (account for castor oil
The 1% of weight), 0.12g lipase G50 (account for castor-oil plant weight of oil 0.4%), then make substrate in 15000rpm down cuts 1min
It is thoroughly mixed, 45 DEG C is then heated under stirring and starts to react, stop heating after reaction, sample in 10000rpm
Lower centrifugation 3min is detached, and upper oil phase is taken to detect acid value.6h acid values are 169.09mgKOH/g, calculate percent hydrolysis and are
92.83%, acid value is 173.40mgKOH/g for 24 hours, and it is 95.22% to calculate percent hydrolysis.
Embodiment 5
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.24g lipase TL and (account for castor-oil plant
The 0.8% of weight of oil), 0.12g lipase G50 (account for castor-oil plant weight of oil 0.4%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 173.47mgKOH/g for 24 hours, calculates hydrolysis
Rate is 95.27%.
Embodiment 6
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 178.04mgKOH/g for 24 hours, calculates hydrolysis
Rate is 97.80%.
Embodiment 7
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.09g lipase TL and (account for castor-oil plant
The 0.3% of weight of oil), 0.06g lipase G50 (account for castor-oil plant weight of oil 0.2%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 180.15mgKOH/g for 24 hours, calculates hydrolysis
Rate is 98.97%.
Embodiment 8
30g castor oil is taken in reactor, then adds in 12g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.06g lipase G50 (account for castor-oil plant weight of oil 0.2%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 172.89mgKOH/g for 24 hours, calculates hydrolysis
Rate is 94.94%.
Embodiment 9
30g castor oil is taken in reactor, then adds in 18g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.06g lipase G50 (account for castor-oil plant weight of oil 0.2%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 173.12mgKOH/g for 24 hours, calculates hydrolysis
Rate is 95.07%.
Embodiment 10
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.9g lipase TL and (account for castor oil
The 3% of weight), 0.3g lipase G50 (account for castor-oil plant weight of oil 1%), then make substrate complete in 15000rpm down cuts 1min
Full mixing, 45 DEG C are then heated under stirring and starts to react, and are stopped heating after reaction, are sampled under 10000rpm
Centrifugation 3min is detached, and upper oil phase is taken to detect acid value.Acid value is 181.00mgKOH/g for 24 hours, calculates percent hydrolysis and is
99.44%.
Embodiment 11
30g castor oil is taken in reactor, then adds in 3g deionized waters, and add in 0.06g lipase TL and (account for castor oil
The 0.2% of weight), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 131.20mgKOH/g for 24 hours, calculates hydrolysis
Rate is 71.82%.
Embodiment 12
30g castor oil is taken in reactor, then adds in 150g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 160.64mgKOH/g for 24 hours, calculates hydrolysis
Rate is 88.15%.
Embodiment 13
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.3g lipase TL and (account for castor oil
The 1% of weight), 0.3g lipase Novozyme 435 (account for castor-oil plant weight of oil 1%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 133.17mgKOH/g for 24 hours, calculates hydrolysis
Rate is 73.16%.
Embodiment 14
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase TL and (account for castor-oil plant
The 0.5% of weight of oil), 0.3g lipase Novozyme 435 (account for castor-oil plant weight of oil 1%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 148.70mgKOH/g for 24 hours, calculates water
Solution rate is 81.25%.
Embodiment 15
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase TL and (account for castor-oil plant
The 0.5% of weight of oil), 0.15g lipase Novozyme 435 (account for castor-oil plant weight of oil 0.5%), then under 15000rpm
Shearing 1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating after reaction,
Sampling centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 143.35mgKOH/g for 24 hours, is counted
It is 78.76% to calculate percent hydrolysis.
Embodiment 16
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase TL and (account for castor-oil plant
The 0.5% of weight of oil), 0.15g lipase Novozyme 435 (account for castor-oil plant weight of oil 0.5%), then under 15000rpm
Shearing 1min is thoroughly mixed substrate, and 35 DEG C are then heated under stirring and starts to react, and stops heating after reaction,
Sampling centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 157.15mgKOH/g for 24 hours, is counted
It is 86.34% to calculate percent hydrolysis.
Embodiment 17
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.15g lipase Novozyme 435 (account for castor-oil plant weight of oil 0.2%), then under 15000rpm
Shearing 1min is thoroughly mixed substrate, and 55 DEG C are then heated under stirring and starts to react, and stops heating after reaction,
Sampling centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 160.71mgKOH/g for 24 hours, is counted
It is 88.30% to calculate percent hydrolysis.
Embodiment 18
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase RM IM
(Lypozyme RM IM, account for the 5% of castor-oil plant weight of oil), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), Ran Hou
15000rpm down cuts 1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, reaction terminates
Stop heating afterwards, sample the centrifugation 3min under 10000rpm and detached, upper oil phase is taken to detect acid value.Acid value is for 24 hours
185.65mgKOH/g, it is 99.39% to calculate percent hydrolysis.
Embodiment 19
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.9g lipase RM IM and (account for castor
The 3% of sesame oil weight), 0.3g lipase G50 (account for castor-oil plant weight of oil 1%), then make bottom in 15000rpm down cuts 1min
Object is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 178.95mgKOH/g for 24 hours, calculates hydrolysis
Rate is 98.32%.
Embodiment 20
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.3g lipase RM IM and (account for castor
The 1% of sesame oil weight), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 168.57mgKOH/g for 24 hours, calculates hydrolysis
Rate is 92.55%.
Embodiment 21
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase RM IM and (account for castor
The 0.2% of sesame oil weight), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 111.53mgKOH/g for 24 hours, calculates hydrolysis
Rate is 60.91%.
Embodiment 22
30g castor oil is taken in reactor, then adds in 3g deionized waters, and add in 0.06g lipase RM IM and (account for castor
The 0.2% of sesame oil weight), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 94.76mgKOH/g for 24 hours, calculates water
Solution rate is 52.06%.
Embodiment 23
30g castor oil is taken in reactor, then adds in 150g deionized waters, and add in 0.06g lipase RM IM and (account for
The 0.2% of castor-oil plant weight of oil), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 97.34mgKOH/g for 24 hours, calculates water
Solution rate is 53.48%.
Embodiment 24
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase RM IM and (account for castor
The 0.2% of sesame oil weight), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 92.18mgKOH/g for 24 hours, calculates water
Solution rate is 50.65%.
Embodiment 25
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.6g lipase RM IM and (account for castor
The 2% of sesame oil weight), 0.3g lipase Novozyme 435 (account for castor-oil plant weight of oil 1%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 155.79mgKOH/g for 24 hours, calculates water
Solution rate is 85.59%.
Embodiment 26
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 1.5g lipase Novozyme
435 (account for castor-oil plant weight of oil 5%), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), then cut under 15000rpm
Cutting 1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating after reaction, takes
Sample centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 102.67mgKOH/g for 24 hours, is calculated
Percent hydrolysis is 56.00%.
Embodiment 27
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.3g lipase Novozyme
435 (account for castor-oil plant weight of oil 1%), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), then cut under 15000rpm
Cutting 1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating after reaction, takes
Sample centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 57.27mgKOH/g for 24 hours, is calculated
Percent hydrolysis is 30.82%.
Embodiment 28
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase Novozyme
435 (account for castor-oil plant weight of oil 0.2%), 0.15g lipase G50 (account for castor-oil plant weight of oil 0.5%), then under 15000rpm
Shearing 1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating after reaction,
Sampling centrifuges 3min under 10000rpm and is detached, and upper oil phase is taken to detect acid value.Acid value is 21.95mgKOH/g for 24 hours, is counted
It is 11.23% to calculate percent hydrolysis.
Embodiment 29
30g coconut oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for coconut
The 0.2% of weight of oil), 0.045g lipase G50 (account for coconut weight of oil 0.15%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 209.58mgKOH/g for 24 hours, calculates hydrolysis
Rate is 86.85%.
Embodiment 30
30g fish oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for fish oil weight
Amount 0.2%), 0.045g lipase G50 (account for fish oil weight 0.15%), then make substrate in 15000rpm down cuts 1min
It is thoroughly mixed, 45 DEG C is then heated under stirring and starts to react, stop heating after reaction, sample in 10000rpm
Lower centrifugation 3min is detached, and upper oil phase is taken to detect acid value.Acid value is 118.49mgKOH/g for 24 hours, calculates percent hydrolysis and is
56.47%.
Embodiment 31
30g purified soyabean oils are taken in reactor, then add in 60g deionized waters, and add in 0.06g lipase TL and (account for
The 0.2% of soybean weight of oil), 0.045g lipase G50 (account for soybean weight of oil 0.15%), then in 15000rpm down cuts
1min is thoroughly mixed substrate, and 45 DEG C are then heated under stirring and starts to react, and stops heating, sampling after reaction
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 166.78mgKOH/g for 24 hours, calculates water
Solution rate is 95.29%.
Embodiment 32
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.075g lipase G50 (account for castor-oil plant weight of oil 0.25%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 174.18mgKOH/g for 24 hours, calculates hydrolysis
Rate is 96.23%.
Embodiment 33
30g lards are taken in reactor, then add in 60g deionized waters, and add in 0.06g lipase TL and (account for castor oil
The 0.2% of weight), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 168.59mgKOH/g for 24 hours, calculates hydrolysis
Rate is 86.46%.
Embodiment 34
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase TL and (account for castor-oil plant
The 5% of weight of oil), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then make in 15000rpm down cuts 1min
Substrate is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 179.97mgKOH/g for 24 hours, calculates hydrolysis
Rate is 98.87%.
Embodiment 35
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for castor-oil plant
The 0.2% of weight of oil), 0.6g lipase G50 (account for castor-oil plant weight of oil 2%), then make bottom in 15000rpm down cuts 1min
Object is thoroughly mixed, and 45 DEG C are then heated under stirring and starts to react, and is stopped heating after reaction, is sampled
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 154.43mgKOH/g for 24 hours, calculates hydrolysis
Rate is 84.70%.
Embodiment 36
30g castor oil is taken in reactor, then adds in 60g deionized waters, and add in 0.15g lipase RM IM and (account for castor
The 5% of sesame oil weight), 0.045g lipase G50 (account for castor-oil plant weight of oil 0.15%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 178.16mgKOH/g for 24 hours, calculates hydrolysis
Rate is 97.86%.
Comparative example 3
30g coconut oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for coconut
The 0.2% of weight of oil), 0.045g lipase 435 (account for coconut weight of oil 0.15%), then in 15000rpm down cuts 1min
Substrate is thoroughly mixed, 45 DEG C are then heated under stirring and starts to react, stops heating after reaction, samples
3min is centrifuged under 10000rpm to be detached, and upper oil phase is taken to detect acid value.Acid value is 149.44mgKOH/g for 24 hours, calculates hydrolysis
Rate is 81.94%.
Comparative example 4
30g fish oil is taken in reactor, then adds in 60g deionized waters, and add in 0.06g lipase TL and (account for fish oil weight
Amount 0.2%), 0.045g lipase 435 (account for fish oil weight 0.15%), then make substrate in 15000rpm down cuts 1min
It is thoroughly mixed, 45 DEG C is then heated under stirring and starts to react, stop heating after reaction, sample in 10000rpm
Lower centrifugation 3min is detached, and upper oil phase is taken to detect acid value.Acid value is 107.81mgKOH/g for 24 hours, calculates percent hydrolysis and is
51.37%.
Comparative example 5
30g lards are taken in reactor, then add in 60g deionized waters, and add in 0.06g lipase TL and (account for lard weight
Amount 0.2%), 0.045g lipase 435 (account for lard weight 0.15%), then make substrate in 15000rpm down cuts 1min
It is thoroughly mixed, 45 DEG C is then heated under stirring and starts to react, stop heating after reaction, sample in 10000rpm
Lower centrifugation 3min is detached, and upper oil phase is taken to detect acid value.Acid value is 124.87mgKOH/g for 24 hours, calculates percent hydrolysis and is
68.31%.
Claims (10)
1. a kind of hydrolysate oil or the method for improving grease hydrolysis rate, which is characterized in that the method includes using lipase (a)
The step of grease hydrolysis being carried out with lipase (b), wherein the lipase (a) is derived from thermophilic fungal category (Thermomyces
Sp. lipase) or the lipase from root Mucor (Rhizomucor sp.), the lipase (b) are partial glycerides
Lipase.
2. the method as described in claim 1, which is characterized in that
The lipase from thermophilic fungal category (Thermomyces sp.) is derived from Thermomyces lanuginosus
The lipase of (Thermomyces languginosus)；Preferably, it is described to be originated from thermophilic fungal category (Thermomyces sp.)
Lipase be Lipozyme TL；And/or
The lipase from root Mucor (Rhizomucor sp.) is derived from Rhizomucor miehei (Rhizomucor
Miehei lipase)；Preferably, the lipase from root Mucor (Rhizomucor sp.) is Lipozyme
RM IM；And/or
It is mould that the partial glyceride lipase is derived from aspergillus (Aspergillus), Penicillium (Penicillium) or horse traction color
The lipase of Pseudomonas (Malassezia)；Preferably, the partial glyceride lipase is derived from penicillium cammenberti
The lipase of (Penicillium camemberti), more preferably lipase G50.
3. method as claimed in claim 1 or 2, which is characterized in that
The additive amount of the lipase (a) be grease weight 0.2~5%, such as 0.2~4%, 0.2~3%, 0.2~2%,
0.2~1%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.5~5%, 0.5~4%, 0.5~3%, 0.5~
2.5%th, 0.5~2%, 0.5~1.5%, 0.5~1%, 0.8~5%, 0.8~4%, 0.8~3%, 1~5%, 1~4% or
1~3%；And/or
The additive amount of the lipase (b) be grease weight 0.15~2.0%, such as 0.15~1.5%, 0.15~1.2%,
0.15~1.0%, 0.15~0.8%, 0.15~0.6%, 0.15~0.5%, 0.15~0.3%, 0.2~1.8%, 0.2~
1.5%th, 0.2~1.0%, 0.2~0.8%, 0.2~0.6%, 0.2~0.5%, 0.3~2.0%, 0.3~1.5%, 0.3~
1.2%th, 0.3~1.0%, 0.5~2.0%, 0.5~1.8%, 0.5~1.5%, 0.5~1.2% or 0.5~1.0%.
4. the method as described in any one of claim 1-3, which is characterized in that
Hydrolysis carries out in presence of water, it is preferable that the mass ratio of grease and water is 1：0.2~5, such as 1：0.2~4,1：0.2
~3,1：0.3~5,1：0.3~4,1：0.3~3,1：0.4~5,1：0.4~4,1：0.4~3,1：0.6~3 or 1：0.6~2
Deng；And/or
Hydrolysis carries out, such as carried out at a temperature of 40~50 DEG C at a temperature of 35~55 DEG C；And/or
The grease is vegetable oil or animal oil, preferably polished fat；Preferably, the grease is selected from：Castor oil, rice
Oil, sunflower oil, palm oil, palm-kernel oil, peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, Purple Perilla Seed Oil, tea-seed oil, palm fibre
Palmitic acid fruit oil, coconut oil, olive oil, oleum theobromatis, tallowseed oil, almond oil, apricot kernel oil, bancoul nuts oil, rubber seed oil, jade
Rice embryo oil, wheat germ oil, sesame seed oil, linseed oil, oenothera seed oil, hazelnut oil, nut oil, grape seed oil, borage seed
Oil, Seabuckthorm Seed Oil, tomato seed oil, pumpkin seed oil, macadimia nut oil, cocoa butter, algae oil, tallow, lard, sheep oil, chicken fat, fish
The arbitrary mixing of the grease of one or more of oil, seal oil, whale oil, dolphin oil and oyster sauce.
5. the method as described in any one of claim 1-4, which is characterized in that
The method includes use be originated from lipase and the partial glyceride lipase of thermophilic fungal category (Thermomyces sp.) into
The step of row grease hydrolysis；Or
The method includes use be originated from lipase and the partial glyceride lipase of root Mucor (Rhizomucor sp.) into
The step of row grease hydrolysis；Preferably, the dosage of the lipase from root Mucor (Rhizomucor sp.) is oil
The 0.8~5% of fat weight, such as 0.8~4%, 0.8~3%, 1~5%, 1~4% or 1~3%, the partial glyceride lipase
The dosage of enzyme is the 0.15~2% of grease weight, such as 0.15~1%, 0.3~2%, 0.3~1.5%, 0.3~1.2% or
0.3~1.0%.
6. the method as described in any one of claim 1-5, which is characterized in that
The mass ratio of grease and water is 1：0.6~3 or 1：0.6~2, it is described to be originated from thermophilic fungal category (Thermomyces sp.)
The additive amount of lipase account for the 0.2~2% of grease weight, the additive amount of the partial glyceride lipase accounts for grease weight
0.15~1%；Or
The mass ratio of grease and water is 1：0.6~3 or 1：0.6~2, it is described from root Mucor (Rhizomucor sp.)
The additive amount of lipase accounts for the 1~5% of grease weight, the additive amount of the partial glyceride lipase account for grease weight 0.15~
1%.
7. partial glyceride lipase or its combination with following lipase (a) or lipase (b) in grease hydrolysis or improve grease
Application in percent hydrolysis or the application in the lipase compositions for preparing for hydrolysate oil or improving grease hydrolysis rate：
The lipase (a) is derived from the lipase of thermophilic fungal category (Thermomyces sp.) or from root Mucor
The lipase of (Rhizomucor sp.)；With
The lipase (b) is partial glyceride lipase.
8. the use as claimed in claim 7, which is characterized in that
The lipase from thermophilic fungal category (Thermomyces sp.) is derived from Thermomyces lanuginosus
The lipase of (Thermomyces languginosus)；Preferably, it is described to be originated from thermophilic fungal category (Thermomyces sp.)
Lipase be Lipozyme TL；And/or
The lipase from root Mucor (Rhizomucor sp.) is derived from Rhizomucor miehei (Rhizomucor
Miehei lipase)；Preferably, the lipase from root Mucor (Rhizomucor sp.) is Lipozyme
RM IM；And/or
It is mould that the partial glyceride lipase is derived from aspergillus (Aspergillus), Penicillium (Penicillium) or horse traction color
The lipase of Pseudomonas (Malassezia)；Preferably, the partial glyceride lipase is derived from penicillium cammenberti
The lipase of (Penicillium camemberti), more preferably lipase G50.
9. a kind of lipase compositions, which is characterized in that the composition contains：
(a) lipase from thermophilic fungal category (Thermomyces sp.) or from root Mucor (Rhizomucor
Sp. lipase)；With
(b) partial glyceride lipase；
Wherein, the lipase in the composition from thermophilic fungal category (Thermomyces sp.) and the partial glyceride fat
The weight ratio of fat enzyme is not 1；
Preferably, the composition contains：
(a) lipase from root Mucor (Rhizomucor sp.)；With
(b) partial glyceride lipase.
10. lipase compositions as claimed in claim 9, which is characterized in that
The lipase from thermophilic fungal category (Thermomyces sp.) is derived from Thermomyces lanuginosus
The lipase of (Thermomyces languginosus)；Preferably, it is described to be originated from thermophilic fungal category (Thermomyces sp.)
Lipase be Lipozyme TL；And/or
The lipase from root Mucor (Rhizomucor sp.) is derived from Rhizomucor miehei (Rhizomucor
Miehei lipase)；Preferably, the lipase from root Mucor (Rhizomucor sp.) is Lipozyme
RM IM；And/or
It is mould that the partial glyceride lipase is derived from aspergillus (Aspergillus), Penicillium (Penicillium) or horse traction color
The lipase of Pseudomonas (Malassezia)；Preferably, the partial glyceride lipase is derived from penicillium cammenberti
The lipase of (Penicillium camemberti), more preferably lipase G50.
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