CN108239578B

CN108239578B - Method for improving hydrolysis rate of grease

Info

Publication number: CN108239578B
Application number: CN201611206801.9A
Authority: CN
Inventors: 李磊; 丛芳; 郭晓峰
Original assignee: Wilmar Shanghai Biotechnology Research and Development Center Co Ltd
Current assignee: Wilmar Shanghai Biotechnology Research and Development Center Co Ltd
Priority date: 2016-12-23
Filing date: 2016-12-23
Publication date: 2022-06-07
Anticipated expiration: 2036-12-23
Also published as: CN108239578A

Abstract

The invention relates to a method for improving the hydrolysis rate of grease. Specifically, the present invention provides a method for hydrolyzing fats and oils or increasing the rate of hydrolysis of fats and oils, which comprises the step of hydrolyzing fats and oils using (a) a lipase derived from a genus Thermomyces sp or a lipase derived from a genus Rhizomucor sp and (b) a partial glyceride lipase. The present invention also provides a lipase composition comprising (a) a lipase derived from the genus Thermomyces or a lipase derived from the genus Rhizomucor; and (b) a partial glyceride lipase; wherein the weight ratio of the lipase derived from Thermomyces to the partial glyceride lipase in the composition is other than 1. The method and the composition can improve the hydrolysis rate of the grease under the condition of saving the production cost, thereby greatly improving the production efficiency.

Description

Method for improving hydrolysis rate of grease

Technical Field

The invention belongs to the field of grease processing technology, and particularly relates to a method for improving the hydrolysis rate of grease

Background

Ricinoleic acid is a commonly used raw material in the oil industry, primarily for the preparation of sebacic acid and 12-hydroxystearic acid. Ricinoleic acid is a hydrolysis product of castor oil in which the fatty acid content of ricinoleic acid is over 90%, much higher than that of other oil species.

At present, the main preparation methods of ricinoleic acid are high-temperature and high-pressure method and enzymatic hydrolysis. Although the high-temperature and high-pressure method has high production efficiency, the method has strict conditions, and when the method is applied to the castor oil hydrolysis process, hydroxyl groups and carboxyl groups in the ricinoleic acid undergo esterification reaction to form polyester, so that the product has a darker color, and the yield of the later-stage 12-hydroxystearic acid preparation is reduced.

Compared with a high-temperature and high-pressure method, the method for hydrolyzing the castor oil by the enzyme method can obtain a high-quality product under mild conditions, and can reduce the generation of waste water and the pollution to the environment. However, enzymatic hydrolysis also has the following problems: the condition of low hydrolysis efficiency generally exists in the hydrolysis process, and the hydrolysis efficiency cannot reach more than 90%; even if the hydrolysis rate can reach 90%, the enzyme adding amount is large, the cost is high, and the production requirement cannot be met.

For example, CN 200710118466.1 describes a method for hydrolyzing castor oil, which uses a combination of lipases (Novozyme 435, Lipozyme RM IM, Lipozyme TL) to hydrolyze, the reaction temperature is controlled at 35-60 ℃, the water-oil molar ratio is 1-50, the reaction time is 10-30h, and the final yield is about 90%. However, the method has the disadvantages of large amount of lipase added in the hydrolysis process, high cost and unsuitability for industrial production.

CN 201080010073.9 describes a method for preparing ricinoleic acid ester by ester exchange. The main process steps are that short-chain alcohol is added into castor oil, and ricinoleate is generated through alcoholysis reaction under the action of lipase.

Yang wei et al (the "castor oil is prepared by using free lipase catalysis", advances in biotechnology 2014(5) 373-378) have main factors influencing the hydrolysis process of castor oil catalyzed by free lipase NS 81006: the temperature, the enzyme dosage, the water dosage and the stirring speed are researched and optimized, and the hydrolysis rate reaches 94.8 percent in 48 hours under the condition of oiling. However, the reaction time of the method is too long, and the production efficiency is influenced in the actual production process.

Puthli M S et al ("Enzymatic hydrolysis of castor oil: Process interaction students", Biochemical Engineering Journal, 2006, 31(1):31-41) for the various factors of Enzymatic hydrolysis of castor oil: factors such as temperature, time, oil-water molar ratio, acetone addition amount, enzyme addition amount, and the like were examined. The article reports that the hydrolysis rate is low, and the normal production requirement cannot be met.

Therefore, there is still a need in the art for a method for hydrolyzing fats and oils by an enzymatic method, which can improve the hydrolysis rate of fats and oils while saving production costs, thereby greatly improving production efficiency.

Disclosure of Invention

The two lipases are compounded, so that the respective characteristics of the two lipases are fully exerted, a high hydrolysis rate can be achieved in a short time under the condition of a small enzyme addition amount, the production efficiency is improved, and the production cost is saved.

Accordingly, in a first aspect, the present invention provides a method for hydrolyzing fats and oils, which comprises the step of hydrolyzing fats and oils using a lipase (a), i.e., a lipase derived from the genus Thermomyces sp or a lipase derived from the genus Rhizomucor sp, and a lipase (b), i.e., a partial glyceride lipase.

In a second aspect, the present invention provides a method for increasing the rate of fat hydrolysis, the method comprising the step of hydrolyzing fat using a lipase (a) derived from a thermophilic fungus (Thermomyces sp) or a Rhizomucor (Rhizomucor sp) lipase, and a lipase (b) which is a partial glyceride lipase.

In one or more embodiments, the lipase derived from the genus Thermomyces (Thermomyces sp.) is a lipase derived from Thermomyces lanuginosus (Thermomyces lanuginosus).

In one or more embodiments, the lipase derived from the genus thermophilic fungus (Thermomyces sp.) is Lipozyme TL.

In one or more embodiments, the lipase derived from Rhizomucor sp is a lipase derived from Rhizomucor miehei (Rhizomucor miehei).

In one or more embodiments, the lipase derived from Rhizomucor sp is Lipozyme RM IM.

In one or more embodiments, the partial glyceride lipase is a lipase derived from penicillium (penicillium sp.).

In one or more embodiments, the lipase derived from Penicillium sp is a lipase derived from Penicillium camemberti (Penicillium camemberti).

In one or more embodiments, the Lipase derived from Penicillium sp is Lipase G50, a commercial enzyme Lipase G "Amano" 50.

In one or more embodiments, the lipase (a) is added in an amount of 0.2 to 5% by weight, for example, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5%, 0.5 to 1%, 0.8 to 5%, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3% by weight of the fat.

In one or more embodiments, the amount of the lipase (b) added is 0.15 to 2.0% by weight of the fat, for example, 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5%, 0.5 to 1.2%, 0.5 to 1.0%.

In one or more embodiments, the hydrolysis is carried out in the presence of water.

In one or more embodiments, the mass ratio of grease to water is 1: 0.2 to 5, for example 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 to 3, etc.

In one or more embodiments, the hydrolysis is carried out at a temperature of from 35 to 55 ℃, for example, from 40 to 50 ℃.

In one or more embodiments, the oil or fat is a vegetable oil or an animal oil.

In one or more embodiments, the oil is a refined oil.

In one or more embodiments, the oil is selected from: castor oil, rice oil, sunflower seed oil, palm kernel oil, peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, perilla seed oil, tea seed oil, palm fruit oil, coconut oil, olive oil, cocoa bean oil, Chinese tallow tree seed oil, almond oil, tung oil, rubber seed oil, corn germ oil, wheat germ oil, sesame seed oil, linseed oil, evening primrose seed oil, hazelnut oil, walnut oil, grape seed oil, glass endive seed oil, sea buckthorn seed oil, tomato seed oil, pumpkin seed oil, macadamia nut oil, cocoa butter, algal oil, beef tallow, lard, mutton fat, chicken fat, fish oil, seal oil, whale oil, dolphin oil and oyster oil.

In one or more embodiments, the oil is castor oil.

In one or more embodiments, the method includes the step of performing lipid hydrolysis using a lipase derived from the genus thermophile (Thermomyces sp.) with a partial glyceride lipase.

In one or more embodiments, the method includes the step of performing oleolysis with a partial glyceride lipase using a lipase derived from rhizomucor (Rhizomucorsp.); preferably, in one or more embodiments, the lipase derived from Rhizomucor sp is used in an amount of 0.8 to 5% by weight of the oil, such as 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3%; preferably, in one or more embodiments, the partial glyceride lipase is used in an amount of 0.15 to 2%, such as 0.15 to 1%, 0.3 to 2%, 0.3 to 1.5%, 0.3 to 1.2%, or 0.3 to 1.0% by weight of the fat.

In a third aspect, the invention provides the use of a partial glyceride lipase or a combination thereof with the following lipases in the hydrolysis of fats and oils or in increasing the rate of hydrolysis of fats and oils:

a lipase (a), i.e., a lipase derived from the genus Thermomyces (Thermomyces sp.) or a lipase derived from the genus Rhizomucor (Rhizomucor sp.); and

lipase (b), i.e., partial glyceride lipase.

In a fourth aspect, the present invention provides a lipase composition comprising:

lipase (b), i.e., partial glyceride lipase;

wherein the weight ratio of lipase derived from Thermomyces sp to the partial glyceride lipase in the composition is other than 1.

In one or more embodiments, the composition contains lipase (a) and lipase (b):

the lipase (a) is a lipase derived from Rhizomucor sp; and

the lipase (b) is a partial glyceride lipase.

In one or more embodiments, the Lipase derived from Penicillium sp is Lipase G50(Lipase G "Amano" 50).

Detailed Description

Lipase enzyme

Herein, "partial glyceride lipase" refers to a lipase which does not have catalytic hydrolysis activity on triglycerides, and preferentially has catalytic hydrolysis activity on diglycerides and/or monoglycerides. Partial glyceride lipases are mainly of two types: monoglyceride-diglyceride lipase and monoglyceride lipase. The present invention preferably uses partial glyceride lipases having catalytic hydrolytic activity for mono-and/or diglycerides. Partial glyceride lipases which are well known in the art may be used, for example, partial glyceride lipases from Aspergillus (Aspergillus), Penicillium (Penicillium) and Malassezia (Malassezia) may be used.

In certain embodiments, partial glyceride lipases suitable for use in the present invention include, but are not limited to, partial glyceride lipases as disclosed in CN 201410182887.0, commercially available partial glyceride lipases such as Lipase G50(Lipase G "Amano"50), and the lipases GH1(Huang J, Yang Z, Guan F et al, alpha novel mono-and diacylglycerol Lipase high expressed in Pichia pastoris, and its application for food emulsion prediction, Process Biochemistry, 2013, 48(12):1899 and 1904), Lipase SMG1 (Xssi T, Lu L, Hou S et al, Crystal lattice of mono-and diacylglycerol Lipase from Lipase front, glycerol front starch front, glycerol front starch front, glycerol front, rear, glycerol front, glycerol rear, and rear, molecular Lipase front, glycerol front, rear, glycerol front starch front, rear starch front starch, rear starch front starch, rear starch front starch, rear starch, rear starch, rear starch, rear starch, rear, biochemical catalysis and molecular basis for substrate selection, Plos One, 2014, 9(7): e102040), and the like.

When used in the method or use of the present invention, the amount of the partial glyceride lipase is usually 0.12 to 2.0% by weight, for example, 0.15 to 2.0%, 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5%, 0.5 to 1.2%, 0.5 to 1.0%, etc., based on the weight of the oil.

In certain embodiments, the present invention uses a partial glyceride lipase from Penicillium sp, particularly from Penicillium camemberti (Penicillium camemberti). For example, the partial glyceride Lipase from Penicillium sp may be Lipase G50, Lipase G "Amano" 50.

In the present invention, another lipase to be compounded with the partial glyceride lipase may be a lipase derived from a genus Thermomyces (Thermomyces sp.) or a lipase derived from a genus Rhizomucor (Rhizomucor sp.). In certain embodiments, the lipase derived from the genus thermophile (Thermomyces sp.) is a lipase derived from Thermomyces lanuginosus (Thermomyces lanuginosus), for example the lipase derived from the genus thermophile (Thermomyces sp.) is Lipozyme TL IM or a lipase derived from the genus thermophile (Thermomyces sp.) having similar enzymatic properties as Lipozyme TL IM.

In certain embodiments, the lipase derived from Rhizomucor sp may be a Rhizomucor miehei (Rhizomucor miehei) derived lipase, e.g., the Rhizomucor sp derived lipase is Lipozyme RM IM or a Rhizomucor derived lipase having similar enzymatic properties as Lipozyme RM IM.

When used in the method or the use of the present invention, the amount of the lipase derived from Thermomyces sp or the lipase derived from Rhizomucor sp may be 0.15 to 5% by weight, for example, 0.2 to 5%, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5%, 0.5 to 1%, 0.8 to 5%, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3% based on the weight of the fat.

In certain embodiments, the present invention uses a lipase derived from Rhizomucor sp in an amount of 0.8 to 5%, such as 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3% by weight of the oil.

In certain embodiments, the present invention uses a combination of a partial glyceride lipase with a lipase derived from the genus thermophilic fungi (Thermomyces sp.). In these embodiments, the partial glyceride lipase may be used in an amount of 0.15 to 2.0% by weight of the fat, for example 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5%, 0.5 to 1.2% or 0.5 to 1.0%; the amount of the lipase derived from Thermomyces sp may be 0.2 to 5% by weight, for example, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5%, 0.5 to 1%, 0.8 to 5%, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4% or 1 to 3% by weight of the oil.

Any combination of the above listed ranges of amounts of the two lipases can be used to practice the invention. For example, the amount of the partial glyceride lipase may be 0.15 to 1.5%, such as 0.15 to 1.0%, based on the weight of the fat, and the amount of the lipase derived from Thermomyces sp used in combination with the partial glyceride lipase may be 0.2 to 5%, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5% or 0.5 to 1% based on the weight of the fat; similarly, the amount of lipase derived from Thermomyces sp may be 0.2 to 5%, such as 0.2 to 2.0%, and the amount of partial glyceride lipase used in combination with the lipase is 0.15 to 2.0%, such as 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2%, 0.5 to 2.0%, 0.2 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5% or 0% by weight of the fat.

In some embodiments, the partial glyceride lipase may be used in an amount of 0.15 to 1%, such as 0.15 to 0.6%, 0.15 to 0.5% or 0.15 to 0.3% by weight of the fat, and the lipase derived from Thermomyces sp used in combination therewith may be used in an amount of 0.2 to 2%, such as 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 2%, 0.5 to 1.5% or 0.5 to 1% by weight of the fat.

In certain embodiments, the present invention uses a combination of a partial glyceride lipase and a lipase derived from Rhizomucor sp. In these embodiments, it is preferred that the partial glyceride lipase is used in an amount of 0.15 to 2% by weight of the fat, for example 0.15 to 1%, 0.3 to 2%, 0.3 to 1.5%, 0.3 to 1.2% or 0.3 to 1.0%; the lipase derived from Rhizomucor sp may be used in an amount of 0.8 to 5% by weight, for example, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4% or 1 to 3% by weight based on the amount of the fat or oil.

The total weight of all lipases added to the oil is usually not more than 7.0%, for example, 0.35 to 7.0%, such as 0.35 to 6.0%, 0.35 to 5.0%, 0.35 to 4.0%, 0.35 to 3.0%, 0.35 to 2.0%, 0.35 to 1.5%, 0.35 to 1.0%, 0.35 to 0.8%, 0.35 to 0.5%, 0.4 to 7.0%, 0.4 to 6.0%, 0.4 to 5.0%, 0.4 to 4.0%, 0.5 to 6.0%, 0.5 to 5.0%, 0.8 to 6.0%, 0.8 to 5.0%, 1.0 to 6.0%, and the like.

In certain embodiments, the weight of lipase derived from Thermomyces sp or from Rhizomucor sp added to the fat is higher than the weight of partial glyceride lipase added to the fat. For example, in these embodiments, the weight of the lipase derived from Thermomyces sp or the lipase derived from Rhizomucor sp added to the oil or fat is at least 1.1 times, such as 1.1 to 33.3 times, for example 1.1 to 30 times, 1.1 to 25 times, 1.1 to 20 times, 1.1 to 15 times, 1.1 to 10 times, etc., the weight of the partial glyceride lipase added to the oil or fat.

In certain embodiments, a lipase derived from a thermophilic fungus (Thermomyces sp.) and a partial glyceride lipase described herein are added to an oil or fat in an amount that is at least 1.2 times, such as at least 1.3 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., the amount of the lipase added to the oil or fat is greater than the amount of the partial glyceride lipase added to the oil or fat. In other embodiments, the partial glyceride lipase is added in an amount of at least 1.1 times, e.g., at least 1.2 times, e.g., 1.1 to 10 times, 1.1 to 8 times, 1.1 to 5 times, 1.1 to 3 times, 1.2 to 8 times, etc., the amount of lipase derived from a genus Thermomyces sp.

In other embodiments, a lipase derived from Rhizomucor sp is added to an oil or fat in an amount that is at least 1.5 times, e.g., at least 2 times, e.g., 2 to 33.3 times, e.g., 1.5 to 30 times, 1.5 to 25 times, 1.5 to 20 times, 1.5 to 15 times, 1.5 to 10 times, 3 to 10 times, etc., greater than the amount of a partial glyceride lipase described herein.

It will be appreciated that in these embodiments, the lipase is added in an amount that, while meeting the fold requirement, also falls within the ranges set forth herein before.

The lipase used in the present invention can be used in various forms well known in the art. For example, the lipase may be a mixture of one or more of liquid enzyme, powdery enzyme, solid enzyme, immobilized enzyme, and the like.

Lipase composition

The present invention also provides a lipase composition comprising (a) a lipase derived from Thermomyces sp or a lipase derived from Rhizomucor sp and (b) a partial glyceride lipase.

The lipase contained in the lipase composition added to the oil or fat is usually not more than 7.0% by weight, for example, 0.35 to 7.0% by weight, such as 0.35 to 6.0%, 0.35 to 5.0%, 0.35 to 4.0%, 0.35 to 3.0%, 0.35 to 2.0%, 0.35 to 1.5%, 0.35 to 1.0%, 0.35 to 0.8%, 0.35 to 0.5%, 0.4 to 7.0%, 0.4 to 6.0%, 0.4 to 5.0%, 0.4 to 4.0%, 0.5 to 6.0%, 0.5 to 5.0%, 0.8 to 6.0%, 0.8 to 5.0%, 1.0 to 6.0% by weight of the oil or fat.

The content of the lipase (a) in the lipase composition of the present invention is sufficient to make the lipase (a) 0.2 to 5% by weight of the oil when the composition is added to the oil in an amount of not more than 7% by weight of the lipase contained, for example, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5%, 0.5 to 1%, 0.8 to 5%, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, 1 to 3%, or the like.

The content of the lipase (b), i.e., partial glyceride lipase, in the lipase composition of the present invention is sufficient that the partial glyceride lipase accounts for 0.15 to 2.0% by weight of the oil when the composition is added to the oil in an amount of not more than 7% by weight of the oil, for example, 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5% or 1.5% by weight of the oil.

In some embodiments, the lipase (a) in the lipase composition is a lipase derived from Thermomyces sp in an amount sufficient such that when the lipase composition is added to an oil in an amount of not more than 7% by weight of the oil as lipase contained, the amount of lipase derived from Thermomyces sp is 0.2 to 5% by weight of the oil, for example, 0.2 to 4%, 0.2 to 3%, 0.2 to 2%, 0.2 to 1%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.5 to 5%, 0.5 to 4%, 0.5 to 3%, 0.5 to 2.5%, 0.5 to 2%, 0.5 to 1.5%, 0.5 to 1%, 0.8 to 5%, 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3% by weight of the oil; the partial glyceride lipase in the lipase (b) is contained in the composition in an amount sufficient that the partial glyceride lipase accounts for 0.15 to 2.0% by weight of the fat when the lipase composition is added to the fat in an amount of not more than 7% by weight of the fat, for example, 0.15 to 1.5%, 0.15 to 1.2%, 0.15 to 1.0%, 0.15 to 0.8%, 0.15 to 0.6%, 0.15 to 0.5%, 0.15 to 0.3%, 0.2 to 1.8%, 0.2 to 1.5%, 0.2 to 1.0%, 0.2 to 0.8%, 0.2 to 0.6%, 0.2 to 0.5%, 0.3 to 2.0%, 0.3 to 1.5%, 0.3 to 1.2%, 0.3 to 1.0%, 0.5 to 2.0%, 0.5 to 1.8%, 0.5 to 1.5% or 0.5 to 1.5% by weight of the fat

In still other embodiments, the lipase (a) in the lipase composition of the present invention is a lipase derived from Rhizomucor sp in an amount sufficient to provide 0.8 to 5%, such as 0.8 to 4%, 0.8 to 3%, 1 to 5%, 1 to 4%, or 1 to 3% by weight of the oil or fat when the lipase composition is added to the oil or fat in an amount such that the lipase does not exceed 7% by weight of the oil or fat; the partial glyceride lipase of the lipase (b) is contained in the lipase composition in an amount sufficient that the partial glyceride lipase accounts for 0.15 to 2% by weight of the fat, for example, 0.15 to 1%, 0.3 to 2%, 0.3 to 1.5%, 0.3 to 1.2% or 0.3 to 1.0% by weight of the fat when the lipase composition is added to the fat in an amount such that the lipase contained in the lipase composition accounts for not more than 7% by weight of the fat.

In certain embodiments, the weight ratio of lipase derived from the genus thermophilic fungus (Thermomyces sp.) to the partial glyceride lipase described herein in the lipase compositions of the present invention is other than 1. In certain embodiments, the lipase derived from the genus Thermomyces (Thermomyces sp.) or the lipase derived from the genus Rhizomucor (Rhizomucor sp.) is present in the lipase compositions of the present invention in an amount by weight greater than the weight of the partial glyceride lipase. For example, in these examples, the weight ratio of lipase derived from Thermomyces sp or from Rhizomucor sp to partial glyceride lipase described herein is 1.1-33.3: 1, for example, 1.1 to 30: 1, 1.1-25: 1, 1.1-20: 1, 1.1-15: 1, 1.1-10: 1, etc.

In certain embodiments, the lipase compositions of the invention comprise a lipase derived from a thermophilic fungus (Thermomyces sp.) and a partial glyceride lipase as described herein, the weight of the former being at least 1.2 times, such as at least 1.3 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, etc., the weight of the latter. In other embodiments, the weight of the partial glyceride lipase is at least 1.1 times, e.g., at least 1.2 times, e.g., 1.1-10 times, 1.1-8 times, 1.1-5 times, 1.1-3 times, 1.2-8 times, etc., the weight of the lipase derived from the genus Thermomyces sp.

In other embodiments, the lipase compositions of the invention comprise a lipase derived from Rhizomucorsp (Rhizomucorsp.) in an amount at least 1.5 times, such as at least 2 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., greater than the weight of a partial glyceride lipase described herein.

It is to be understood that in these embodiments, the weight of lipase in the lipase composition, while meeting the fold requirement, should be within the ranges set forth herein above.

The lipase composition of the present invention can be in the form of various lipase compositions known in the art, including but not limited to a liquid composition, a solid composition (e.g., a powder composition), or in the form of an immobilized enzyme.

Hydrolysis process

The lipase or the composition thereof can be used for hydrolyzing the grease, or can be used for improving the hydrolysis rate of the grease.

The fats and oils suitable for use in the method and use of the present invention may be any of various fats and oils known in the art, including vegetable oils or animal oils. Preferably, the fats and oils used in the present invention are liquid at the hydrolysis temperature of the present invention. The oil and fat may be refined oil and fat. For example, suitable fats and oils for use in the present invention may be selected from: castor oil, rice oil, sunflower seed oil, palm kernel oil, peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, perilla seed oil, tea seed oil, palm fruit oil, coconut oil, olive oil, cocoa bean oil, Chinese tallow tree seed oil, almond oil, tung oil, rubber seed oil, corn germ oil, wheat germ oil, sesame seed oil, linseed oil, evening primrose seed oil, hazelnut oil, walnut oil, grape seed oil, glass endive seed oil, sea buckthorn seed oil, tomato seed oil, pumpkin seed oil, macadamia nut oil, cocoa butter, algal oil, beef tallow, lard, mutton fat, chicken fat, fish oil, seal oil, whale oil, dolphin oil and oyster oil. In one or more embodiments, the oil is castor oil.

In the hydrolysis, an appropriate amount of water is added to the oil or fat, depending on the enzyme, oil or fat, and the like to be used. Generally, the mass ratio of grease to water is 1: 0.2 to 5, for example 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 to 2, etc. For optimal hydrolysis, one skilled in the art can determine and adjust the mass ratio of oil to water based on the specific oil used, the amount of enzyme used, the temperature of the reaction, the time of the reaction, and the like.

The lipase may then be added to the oil. It is to be understood that the above-mentioned lipases (a) and (b) may be added to fats and oils, respectively, or may be added in the form of a mixture, or the lipase composition of the present invention may be added to fats and oils. The amount of lipase added is as described above. After the addition, the oil and fat, lipase and water are mixed uniformly. For example, it may be mixed by high speed shearing. If the lipase is added to the fat or oil as an aqueous solution, it is preferable to ensure that the mass ratio of the total amount of water in the aqueous solution of fatty acid to the additionally added water to the fat or oil is within the above range.

The hydrolysis reaction may be carried out at a temperature of 35 to 55 ℃ in general, for example, at a temperature of 40 to 50 ℃. Therefore, after the oil and fat, the lipase and the water are uniformly mixed, the temperature of the obtained mixture is gradually increased to be within the range of 35-55 ℃ for reaction. Usually, the temperature is raised with stirring. The stirring can be continuously carried out in the reaction process.

The time of the hydrolysis reaction is not particularly limited, and may be adjusted depending on the amount of the fat or oil to be hydrolyzed, the kind of the lipase to be used, the amount thereof, and the like. Or, the acid value of the sample can be analyzed and the hydrolysis rate can be calculated in the hydrolysis reaction process, and the reaction can be stopped when the expected hydrolysis rate is reached.

In addition, the amount of each enzyme, the amount of water, the temperature and time of hydrolysis, etc. may be adjusted appropriately according to the type of fat or oil to achieve an optimum hydrolysis rate. In certain embodiments, at least 90% hydrolysis, preferably at least 92% hydrolysis, more preferably at least 95% hydrolysis, more preferably at least 97% hydrolysis, more preferably at least 98% hydrolysis can be achieved using the process of the present invention.

Therefore, the method for hydrolyzing fats and oils or the method for increasing the rate of hydrolysis of fats and oils according to the present invention includes a step of hydrolyzing fats and oils using (a) a lipase derived from a genus Thermomyces (Thermomyces sp.) or a lipase derived from a genus Rhizomucor (Rhizomucor sp.) and (b) a partial glyceride lipase. In certain embodiments, the method comprises blending the oil, adding the mixture of lipase and water, heating to 35-55 ℃, and reacting at that temperature for a period of 6 hours, 8 hours, 12 hours, 16 hours, 24 hours, or more, or a combination thereof, to hydrolyze the oil.

In certain embodiments, the hydrolysis rate of the fats and oils can be increased to more than 90% by the method of the present invention. In certain embodiments, the method of the present invention can increase the hydrolysis rate of fats and oils to more than 92%. The method can improve the hydrolysis rate of the grease to more than 95 percent. The method can improve the hydrolysis rate of the grease to more than 97 percent, preferably to more than 98 percent.

In certain embodiments, using the methods of the present invention, the mass ratio of grease to water is controlled to be 1: 0.6-3 or 1: 0.6-2% of lipase from thermophilic fungi (Thermomyces sp.) accounting for 0.2-2% of the weight of the grease, 0.15-1% of lipase (b) accounting for the weight of the grease, and after a proper reaction time, the hydrolysis rate of the grease can be improved to 95% or more than 98%.

In certain embodiments, using the methods of the present invention, the mass ratio of grease to water is controlled to be 1: 0.6-3 or 1: 0.6-2, the addition amount of the lipase (b) derived from Rhizomucor sp accounts for 1-5% of the weight of the grease, and the addition amount of the lipase (b) accounts for 0.15-1% of the weight of the grease, and after a proper reaction time, the hydrolysis rate of the grease can be improved to 95% or more or 98% or more.

Applications of

The invention also provides application of the partial glyceride lipase in hydrolyzing the oil and the application in preparing the lipase composition for hydrolyzing the oil. Also provided is the use of (a) a lipase derived from Thermomyces sp or a lipase derived from Rhizomucor sp with (b) a partial glyceride lipase in the hydrolysis of fats and oils, and in the preparation of a lipase composition for the hydrolysis of fats and oils. The lipase of the genus Thermomyces (Thermomyces sp.) or the lipase derived from Rhizomucor sp and partial glyceride lipase can be as described above.

The present invention will be illustrated below by way of specific examples. It should be understood that these examples are illustrative only and are not intended to limit the scope of the present invention. The lipases Novozyme 435, Lipozyme RM IM and Lipozyme TL IM used in the examples are products of Novozyme (China) investment Limited, and the Lipase G50 is Lipase G "Amano"50, a product of Amano Enzyme preparation Inc. The castor oil used in the examples was purchased from Fengyi Fine chemistry (Liandong harbor) Inc.; coconut oil was purchased from gali specialty fats & oils (shanghai) ltd; fish oil was purchased from basf (china) ltd under the trade name DHA fish oil; the refined soybean oil is purchased from Shanghai Jiali grain oil Co., Ltd, and other conventional chemicals are purchased from Chinese medicinal chemical reagent Co., Ltd, and are of analytical pure grade. The percentages in the examples are by weight. Lipase TL is added as an aqueous solution by first dissolving Lipozyme TL IM in water to form an aqueous solution of conventional concentration. The other lipases are all added in powder form.

In the examples, the hydrolysis ratio was calculated by the following formula:

wherein, AV₀Refers to the acid value of the grease raw material, which is 1.70mgKOH/g for castor oil; AV (Audio video)_tMeans sampling acid value at a certain time interval t; SV refers to the saponification value of the grease raw material, which for castor oil is 182.01 mgKOH/g.

Comparative example 1

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 1.5g of lipase TL (Lipozyme TL IM, accounting for 5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after 6h of reaction, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, detecting the acid value to be 116.69mgKOH/g, and calculating the hydrolysis rate to be 63.77%.

Comparative example 2

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.6G of Lipase G50(Lipase G 'Amano' 50 accounting for 2% of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after 6h of reaction, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, detecting the acid value to be 53.06mgKOH/G, and calculating the hydrolysis rate to be 28.46%.

Example 1

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 1.5G of lipase TL (5 percent of the weight of the castor oil), 0.6G of lipase G50 (2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after 6h of reaction, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase to detect the acid value to be 177.25mgKOH/G, and calculating the hydrolysis rate to be 97.36 percent.

Example 2

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 1.5G of lipase TL (accounting for 5 percent of the weight of the castor oil), 0.45G of lipase G50 (accounting for 1.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after 8h of reaction, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase to detect the acid value to be 173.61mgKOH/G, and calculating the hydrolysis rate to be 95.34 percent.

Example 3

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 1.5G of lipase TL (5 percent of the weight of the castor oil), 0.3G of lipase G50 (1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after 8 hours of reaction, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase to detect the acid value to be 169.95mgKOH/G, and calculating the hydrolysis rate to be 93.31 percent.

Example 4

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.3G of lipase TL (accounting for 1 percent of the weight of the castor oil), 0.12G of lipase G50 (accounting for 0.4 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 6h was 169.09mgKOH/g, the calculated hydrolysis rate was 92.83%, the acid value of 24h was 173.40mgKOH/g, and the calculated hydrolysis rate was 95.22%.

Example 5

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.24G of lipase TL (accounting for 0.8 percent of the weight of the castor oil) and 0.12G of lipase G50 (accounting for 0.4 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value was 173.47mgKOH/g for 24h, the calculated hydrolysis was 95.27%.

Example 6

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 178.04mgKOH/g, and the calculated hydrolysis rate was 97.80%.

Example 7

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.09G of lipase TL (accounting for 0.3 percent of the weight of the castor oil), 0.06G of lipase G50 (accounting for 0.2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 180.15mgKOH/g, and the calculated hydrolysis rate was 98.97%.

Example 8

Taking 30G of castor oil into a reactor, adding 12G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil) and 0.06G of lipase G50 (accounting for 0.2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 172.89mgKOH/g, and the calculated hydrolysis rate was 94.94%.

Example 9

Taking 30G of castor oil into a reactor, adding 18G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.06G of lipase G50 (accounting for 0.2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value in 24 hours was 173.12mgKOH/g, the calculated hydrolysis was 95.07%.

Example 10

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.9G of lipase TL (accounting for 3 percent of the weight of the castor oil), 0.3G of lipase G50 (accounting for 1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value was 181.00mgKOH/g for 24h, the calculated hydrolysis was 99.44%.

Example 11

Taking 30G of castor oil into a reactor, adding 3G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value for 24h was 131.20mgKOH/g, the calculated hydrolysis was 71.82%.

Example 12

Taking 30G of castor oil into a reactor, adding 150G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h is 160.64mgKOH/g, and the calculated hydrolysis rate is 88.15 percent.

Example 13

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.3g of lipase TL (accounting for 1 percent of the weight of the castor oil) and 0.3g of lipase Novozyme 435 (accounting for 1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value after 24 hours was 133.17mgKOH/g, and the calculated hydrolysis rate was 73.16%.

Example 14

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.15g of lipase TL (accounting for 0.5 percent of the weight of the castor oil), 0.3g of lipase Novozyme 435 (accounting for 1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value after 24 hours was 148.70mgKOH/g, and the calculated hydrolysis rate was 81.25%.

Example 15

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.15g of lipase TL (accounting for 0.5 percent of the weight of the castor oil) and 0.15g of lipase Novozyme 435 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min, separating, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 143.35mgKOH/g, and the calculated hydrolysis rate was 78.76%.

Example 16

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.15g of lipase TL (accounting for 0.5 percent of the weight of the castor oil) and 0.15g of lipase Novozyme 435 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 35 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 157.15mgKOH/g, and the calculated hydrolysis rate was 86.34%.

Example 17

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.15g of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.15g of lipase Novozyme 435 (accounting for 0.2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 55 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 160.71mgKOH/g, and the calculated hydrolysis rate was 88.30%.

Example 18

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 1.5G of lipase RM IM (lysozyme RM IM, accounting for 5 percent of the weight of the castor oil) and 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value was 185.65mgKOH/g for 24h, the calculated hydrolysis was 99.39%.

Example 19

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.9G of lipase RM IM (accounting for 3 percent of the weight of the castor oil), 0.3G of lipase G50 (accounting for 1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 178.95mgKOH/g, and the calculated hydrolysis rate was 98.32%.

Example 20

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.3G of lipase RM IM (accounting for 1 percent of the weight of the castor oil), 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 168.57mgKOH/g, and the calculated hydrolysis rate was 92.55%.

Example 21

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase RM IM (accounting for 0.2 percent of the weight of the castor oil), 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value for 24h was 111.53mgKOH/g, and the calculated hydrolysis rate was 60.91%.

Example 22

Taking 30G of castor oil into a reactor, adding 3G of deionized water, adding 0.06G of lipase RM IM (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 94.76mgKOH/g, and the calculated hydrolysis rate was 52.06%.

Example 23

Taking 30G of castor oil into a reactor, adding 150G of deionized water, adding 0.06G of lipase RM IM (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 97.34mgKOH/g, and the calculated hydrolysis rate was 53.48%.

Example 24

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase RM IM (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 92.18mgKOH/g, and the calculated hydrolysis rate was 50.65%.

Example 25

Taking 30g of castor oil into a reactor, adding 60g of deionized water, adding 0.6g of lipase RM IM (accounting for 2 percent of the weight of the castor oil), 0.3g of lipase Novozyme 435 (accounting for 1 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 155.79mgKOH/g, and the calculated hydrolysis rate was 85.59%.

Example 26

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 1.5G of lipase Novozyme 435 (accounting for 5 percent of the weight of the castor oil) and 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix the substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 102.67mgKOH/g, and the calculated hydrolysis rate was 56.00%.

Example 27

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.3G of lipase Novozyme 435 (accounting for 1 percent of the weight of the castor oil) and 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value for 24h was 57.27mgKOH/g, the calculated hydrolysis was 30.82%.

Example 28

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase Novozyme 435 (accounting for 0.2 percent of the weight of the castor oil) and 0.15G of lipase G50 (accounting for 0.5 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix the substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value was 21.95mgKOH/g for 24h, and the calculated hydrolysis rate was 11.23%.

Example 29

30G of coconut oil is taken in a reactor, 60G of deionized water is added, 0.06G of lipase TL (accounting for 0.2 percent of the weight of the coconut oil) and 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the coconut oil) are added, then the mixture is sheared for 1min at 15000rpm to completely mix the substrates, then the mixture is heated to 45 ℃ under the stirring state to start reaction, the heating is stopped after the reaction is finished, a sample is centrifuged for 3min at 10000rpm to be separated, and the upper oil phase is taken out to detect the acid value. The acid value of 24h was 209.58mgKOH/g, and the calculated hydrolysis rate was 86.85%.

Example 30

Taking 30G of fish oil in a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the fish oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the fish oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 118.49mgKOH/g, and the calculated hydrolysis rate was 56.47%.

Example 31

Taking 30G of refined soybean oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the soybean oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the soybean oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 166.78mgKOH/g, and the calculated hydrolysis rate was 95.29%.

Example 32

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.075G of lipase G50 (accounting for 0.25 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value for 24h was 174.18mgKOH/g, the calculated hydrolysis was 96.23%.

Example 33

Taking 30G of lard oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 168.59mgKOH/g, and the calculated hydrolysis rate was 86.46%.

Example 34

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.15G of lipase TL (accounting for 5 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 179.97mgKOH/g, and the calculated hydrolysis rate was 98.87%.

Example 35

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.06G of lipase TL (accounting for 0.2 percent of the weight of the castor oil), 0.6G of lipase G50 (accounting for 2 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 154.43mgKOH/g, and the calculated hydrolysis rate was 84.70%.

Example 36

Taking 30G of castor oil into a reactor, adding 60G of deionized water, adding 0.15G of lipase RM IM (accounting for 5 percent of the weight of the castor oil), 0.045G of lipase G50 (accounting for 0.15 percent of the weight of the castor oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min to separate, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 178.16mgKOH/g, and the calculated hydrolysis rate was 97.86%.

Comparative example 3

30g of coconut oil is taken in a reactor, 60g of deionized water is added, 0.06g of lipase TL (accounting for 0.2 percent of the weight of the coconut oil) and 0.045g of lipase 435 (accounting for 0.15 percent of the weight of the coconut oil) are added, then the substrates are completely mixed by shearing at 15000rpm, then the substrates are heated to 45 ℃ under the stirring state to start the reaction, the heating is stopped after the reaction is finished, a sample is taken and centrifuged at 10000rpm for 3min for separation, and the upper oil phase is taken out to detect the acid value. The acid value of 24h was 149.44mgKOH/g, and the calculated hydrolysis rate was 81.94%.

Comparative example 4

Taking 30g of fish oil in a reactor, adding 60g of deionized water, adding 0.06g of lipase TL (accounting for 0.2 percent of the weight of the fish oil) and 0.045g of lipase 435 (accounting for 0.15 percent of the weight of the fish oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value after 24 hours was 107.81mgKOH/g, and the calculated hydrolysis rate was 51.37%.

Comparative example 5

Taking 30g of lard oil into a reactor, adding 60g of deionized water, adding 0.06g of lipase TL (accounting for 0.2 percent of the weight of the lard oil), 0.045g of lipase 435 (accounting for 0.15 percent of the weight of the lard oil), shearing at 15000rpm for 1min to completely mix substrates, heating to 45 ℃ under a stirring state to start reaction, stopping heating after the reaction is finished, sampling, centrifuging at 10000rpm for 3min for separation, taking an upper oil phase, and detecting the acid value. The acid value of 24h was 124.87mgKOH/g, and the calculated hydrolysis rate was 68.31%.

Claims

1. A method for hydrolyzing fats and oils or increasing the hydrolysis rate of fats and oils, comprising the step of hydrolyzing fats and oils using a lipase (a) and a lipase (b), wherein the lipase (a) is derived from the genus Thermomyces (Thermomyces: (A))Thermomyces sp.) The lipase Lipozyme TL of (1) or derived from Rhizomucor genus (A), (B)Rhizomucor sp.) The lipase Lipozyme RM IM of (a), wherein the lipase (b) is derived from Penicillium (A)Penicillium sp.) Partial glyceride lipase G50; wherein the addition amount of the lipase (a) is 0.15-5% of the weight of the grease, and the addition amount of the lipase (b) is 0.12-2.0% of the weight of the grease.

2. The method of claim 1,

the addition amount of the lipase (a) is 0.2-5% of the weight of the grease; and/or

The addition amount of the lipase (b) is 0.15-2.0% of the weight of the grease.

3. The method according to claim 2, wherein the lipase (a) is added in an amount of 0.2 to 3% by weight based on the fat or oil.

4. The method according to claim 2, wherein the lipase (a) is added in an amount of 0.2 to 2% by weight based on the fat or oil.

5. The method according to claim 2, wherein the lipase (a) is added in an amount of 0.2 to 1% by weight based on the fat or oil.

6. The method according to claim 2, wherein the lipase (a) is added in an amount of 0.2 to 0.8% by weight based on the fat or oil.

7. The method according to claim 2, wherein the lipase (a) is added in an amount of 0.5 to 3% by weight based on the fat or oil.

8. The method according to claim 2, wherein the amount of the lipase (b) added is 0.15 to 1.5% by weight based on the fat or oil.

9. The method according to claim 2, wherein the amount of the lipase (b) added is 0.15 to 1.0% by weight based on the fat or oil.

10. The method according to claim 2, wherein the amount of the lipase (b) added is 0.15 to 0.5% by weight based on the fat or oil.

11. The method according to claim 2, wherein the amount of the lipase (b) added is 0.2 to 1.0% by weight based on the fat or oil.

12. The method of claim 1,

the hydrolysis is carried out in the presence of water; and/or

The hydrolysis is carried out at the temperature of 35-55 ℃; and/or

The oil is vegetable oil or animal oil.

13. The method of claim 12, wherein the mass ratio of grease to water is 1: 0.2 to 5.

14. The method of claim 13, wherein the mass ratio of grease to water is 1: 0.2 to 3.

15. The method of claim 12, wherein the hydrolysis is carried out at a temperature of 40 to 50 ℃.

16. The method of claim 12, wherein the oil is refined oil.

17. The method of claim 12, wherein the grease is selected from the group consisting of: castor oil, rice oil, sunflower seed oil, palm kernel oil, peanut oil, rapeseed oil, cottonseed oil, safflower seed oil, perilla seed oil, tea seed oil, palm fruit oil, coconut oil, olive oil, cocoa bean oil, Chinese tallow tree seed oil, almond oil, tung oil, rubber seed oil, corn germ oil, wheat germ oil, sesame seed oil, linseed oil, evening primrose seed oil, hazelnut oil, walnut oil, grape seed oil, glass endive seed oil, sea buckthorn seed oil, tomato seed oil, pumpkin seed oil, macadamia nut oil, cocoa butter, algal oil, beef tallow, lard, mutton fat, chicken fat, fish oil, seal oil, whale oil, dolphin oil and oyster oil.

18. The method of claim 17, wherein the oil is castor oil.

19. The method of any one of claims 1-2 and 12-18,

the method comprises using a microorganism derived from the genus Thermomyces (A)Thermomyces sp.) The lipase Lipozyme TL and the partial glyceride lipase G50 are subjected to grease hydrolysis; or

The method comprises using a microorganism derived from Rhizomucor genus (A), (B), (C) and D) a (C)Rhizomucor sp.) The lipase Lipozyme RM IM (1) and the partial glyceride lipase G50 are subjected to a step of hydrolyzing fats and oils.

20. The method of claim 19, wherein said source is from rhizomucor genus(s) (c: (a) ((b))Rhizomucor sp.) The amount of the lipase Lipozyme RM IM is 0.8-5% of the weight of the grease, and the amount of the partial glyceride lipase G50 is 0.15-2% of the weight of the grease.

21. As claimed in claim 20The method is characterized in that the microorganism is derived from Rhizomucor rhizopus (A), (B), (C) and (C)Rhizomucor sp.) The amount of the lipase Lipozyme RM IM is 0.8-3% of the weight of the grease, and the amount of the partial glyceride lipase G50 is 0.15-1% of the weight of the grease.

22. The method of claim 20, wherein the source is from rhizomucor genus(s) (c: (a) ((b))Rhizomucor sp.) The amount of the lipase Lipozyme RM IM is 0.8-3% of the weight of the grease, and the amount of the partial glyceride lipase G50 is 0.3-1.5% of the weight of the grease.

23. The method of any one of claims 12-18,

the mass ratio of the grease to the water is 1: 0.6-3 or 1: 0.6 to 2, derived from the genus Thermomyces: (Thermomyces sp.) The addition amount of the lipase Lipozyme TL accounts for 0.2-2% of the weight of the grease, and the addition amount of the partial glyceride lipase G50 accounts for 0.15-1% of the weight of the grease; or

The mass ratio of the grease to the water is 1: 0.6-3 or 1: 0.6 to 2, derived from Rhizomucor genus (A)Rhizomucor sp.) The addition amount of the lipase Lipozyme RM IM is 1-5% of the weight of the grease, and the addition amount of the partial glyceride lipase G50 is 0.15-1% of the weight of the grease.

24. From the genus Thermomyces (A), (B), (C)Thermomyces sp.) The lipase Lipozyme TL of (1) or derived from Rhizomucor genus (A), (B)Rhizomucor sp.) Lipozyme RM IM and a lipase derived from Penicillium (A)Penicillium sp.) The use of the partial glyceride lipase G50 in oil and fat hydrolysis or oil and fat hydrolysis rate improvement, or the use in the preparation of a lipase composition for oil and fat hydrolysis or oil and fat hydrolysis rate improvement, wherein, in the use, the partial glyceride lipase G50 is derived from thermophilic fungus (Thermomyces sp.) (AThermomyces sp.) The lipase Lipozyme TL of (1) or derived from Rhizomucor genus (A), (B)Rhizomucor sp.) The addition amount of the lipase Lipozyme RM IM is 0.15-5% of the weight of the grease, and the addition amount of the partial glyceride lipase G50 is 0.12-2.0% of the weight of the grease.

25. The use according to claim 24, wherein said source is from the genus thermophilic fungi (M.), (Thermomyces sp.) The lipase Lipozyme TL of (1) or derived from Rhizomucor genus (A), (B)Rhizomucor sp.) The lipase Lipozyme RM IM is added in the amount of any one of claims 2 to 7, and the partial glyceride lipase G50 is added in the amount of any one of claims 2 and 8 to 11.