Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Reexamination Certificate
1999-11-24
2001-07-10
Naff, David M. (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
C435S177000, C435S180000, C435S289100
Reexamination Certificate
active
06258575
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a process for efficiently hydrolyzing fats and oils to produce aliphatic acids. In particular, the present invention relates to a process for efficiently hydrolyzing fats and oils without the reduction of enzyme activity caused by an increase in the number of treatments. In the description, the term of “fats and oils” may mean an inclusion of a fat, an oil, a lard, a grease, and so on.
2. Description of the Background
As continuous processes for hydrolyzing fats and oils by an enzyme, there are processes wherein an lipolytic enzyme (lipase) is immobilized onto resin or membrane and used in a circulating reactor such as a batch stirring multi-stage reactor, a packed column or a membrane reactor and an oily substrate and a water-soluble substrate are circulated in countercurrent or cocurrent (JP-A 61-85195, JP-A 63-59896 and JP-A 1-98494). In any methods, however, apparatuses are complicated and the method of operating an enzyme column is very difficult.
As reaction examples in a batch circulating system, e.g. the following techniques are known: (1) a method of conducting a mixture having fats and oils mixed with water in an amount of about 60 weight % relative to the fats and oils to pass through an enzyme-packed reactor; (2) a method of conducting only fats and oils not containing an aqueous phase substrate to pass through a lipase immobilized on polysaccharide gel containing a large amount of water (JP-A 58-146284); (3) a method of mixing fats and oils with water uniformly by circulating them through a fat- and oil-storing tank and a humidifying apparatus and then conducting the mixture with the water content kept constant to pass through an enzyme-packed reactor (JP-A 4-335881); and (4) a method of introducing fats, oils, and water to a substrate-circulating chamber, feeding the oil phase substrate only to an enzyme column while the oil phase is separated from the aqueous phase, and returning the discharged reaction solution to the bottom of the substrate-circulating chamber (Kosugi, Y., Tomizuka, N., J. Am. Oil. Chem. Soc. 72;1329 (1995)).
In the process for hydrolyzing fats and oils by an enzyme, it is desired that the enzyme is not lost while attaining a sufficient reaction rate in easy operation in simple facilities and also that an oil phase product and an aqueous phase product can be separated from each other without centrifugation or a membrane.
Further, the above-described method (1) has the problem that because the enzyme is removed with time, the enzyme activity is lowered as the number of treatments is increased, although the degree of hydrolysis is high due to a high water content in the mixture. In the method (2) unlike the method (1), the enzyme is not removed with time, but water being necessary for hydrolysis tends to be insufficient while glycerol is accumulated in the immobilized enzyme, thus shifting the equilibrium to the side of fats and oils, which may result in inadequate decomposition of fats and oils. In the method (3), separation of the oil phase from the aqueous phase after decomposition (or hydrolysis) is difficult thus making a separation step and facilities therefor necessary and incidental. Further, any method of the methods (1) to (3) suffers from the problem that the apparatuses are complicated or the enzyme-packed reactor is very difficult to operate. In the method (4), the water content in the oil phase passing through the enzyme reactor is low and the separation of the oil phase from the aqueous phase after decomposition (or hydrolysis) is easy, but there is the problem that it is difficult to obtain high degrees of decomposition due to inadequate extraction of glycerol from the reaction solution.
SUMMARY OF THE INVENTION
The present inventors have found a process in which an immobilized enzyme is used for recovery of the enzyme and while the oil phase and the aqueous phase are separated from each other naturally in a substrate-feeding chamber, the reaction is repeatedly conducted thereby solving the conventionally problematic reaction rate and separation of products, and further because the apparatuses consisting of only an enzyme column and a substrate-feeding chamber, the facilities are simple and operation are used easily to be able to achieve stable operation, and simultaneously the investment for the facilities are economical.
Another object of the present invention is to provide a process for efficiently hydrolyzing fats and oils in easy operation at high degrees of hydrolysis without lowering enzyme activity with time.
In addition, the present inventors have found that after the oil and water are separated in the substrate-feeding chamber, the water content in the oil phase substrate fed to the enzyme column is made up to have sufficient amount whereby the hydrolysis reaction in the enzyme column is promoted, and thereafter the reaction solution is brought into contact with a sufficient amount of water whereby the glycerol formed by hydrolysis is dissolved so that the reaction is prevented from being shifted to the side of fats and oils, whereby the hydrolysis can be conducted sufficiently, the apparatus used is made simpler, and the operation is made easier.
The present invention relates to a process for hydrolyzing fats and oils, which comprises using an enzyme column (reaction column) using an immobilized enzyme and a substrate-feeding chamber also serving as an oil-water separating chamber, circulating a reaction solution between the enzyme column and the substrate-feeding chamber and simultaneously separating oils from water in the substrate-feeding chamber, wherein an oil phase substrate and an aqueous phase substrate are removed separately without previously mixing them in the substrate-feeding chamber, and then the hydrolysis of fats and oils is conducted in any of the following processes:
1) a process for hydrolyzing fats and oils, which comprises mixing an oil phase substrate with an aqueous phase substrate and feeding the mixture to the enzyme column;
2) a process for hydrolyzing fats and oils, which comprises mixing an oil phase substrate with an aqueous phase substrate, feeding the mixture to the enzyme column, and introducing a reaction solution having passed through the enzyme column into a mixing chamber for bringing the reaction solution into contact with an aqueous phase substrate, and bringing the reaction solution into contact with the aqueous phase substrate in said mixing chamber, while, after that, returning the mixture to the substrate-feeding chamber; and
3) a process for hydrolyzing fats and oils, which comprises feeding only an oil phase substrate to the enzyme column, introducing a reaction solution having passed through the enzyme column into a mixing chamber for bringing the reaction solution into contact with an aqueous phase substrate and bringing the reaction solution into contact with the aqueous phase substrate in said mixing chamber, while, after that, returning the mixture to the substrate-feeding chamber.
Preferably, an oil-water interface can be formed during reaction in the substrate-feeding chamber. The water content in the oil phase substrate after separation of oils and water is preferably from the saturated one to 5%.
The aqueous phase substrate can be fed from the substrate-feeding chamber to the oil-water mixing chamber.
Preferably, the reaction solution is brought into contact with the aqueous phase substrate by stirring or in a countercurrent contacting system in the oil-water mixing chamber.
In addition, the present invention comprises conducting an oil phase and an aqueous phase to be separated in the feeding chamber without stirring, removing them through respective pipes to bring them into contact with an enzyme, separating the reaction solution flowing from the enzyme column into an oil phase and an aqueous phase in the feeding chamber, and repeating this operation-series, to solve the problem described above. In the present invention, the pipe for returning the reaction solution to the substr
Komatsu Toshiteru
Shimizu Masami
Yamada Naoto
Kao Corporation
Naff David M.
Oblon & Spivak, McClelland, Maier & Neustadt P.C.
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