Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Stablizing an enzyme by forming a mixture – an adduct or a...
Patent
1996-07-25
2000-02-29
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Stablizing an enzyme by forming a mixture, an adduct or a...
435193, 426 20, C12N 900
Patent
active
060308214
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a stable transglutaminase having excellent shelf life (as the case may be, transglutaminase is abbreviated as TGase hereinafter) and a transglutaminase enzyme preparation which contains it as an active ingredient.
BACKGROUND ART
Decreased enzyme activities are observable in many enzymes when stored for a prolonged period of time.
TGase is an enzyme whose enzyme activity is decreased in a marked degree due, mainly, to oxygen when stored for a prolonged period of time. In consequence, a process in which an organic acid, an inorganic acid, a polyphenol, a thiol compound, a sugar alcohol and the like are used as additives has been developed with the aim of improving shelf life of TGase (cf. Japanese Patent Application Kokai No. 4-207194). However, improvement of shelf life by these additives is not sufficient. Especially, when TGase is stored under severe conditions such as storage throughout the summer season after its production, it is necessary to provide a proper means such as the use of an oxygen scavenger or vacuum treatment of the packaging, but these means are also still unsatisfactory.
DISCLOSURE OF THE INVENTION
In view of the above, it therefore becomes an object of the present invention to provide a stabilized TGase which can be stored for a prolonged period of time at ordinary temperature without requiring oxygen scavenger, vacuum packaging and the like and a transglutaminase enzyme preparation which contains the same.
With the aim of achieving the aforementioned object, the inventors of the present invention have conducted intensive studies and found that treatment of TGase by a specified method using a specified substance can stabilize it markedly so that its enzyme activity does not decrease even after a prolonged period of storage at ordinary temperature, and have accomplished the present invention on the basis of such a finding.
Accordingly, the present invention relates to a stabilized TGase obtained by drying a solution containing TGase and a protein material and to an enzyme preparation which contains it as an active ingredient.
The following illustratively describes the present invention.
Origin of the TGase to be stabilized in the present invention is not particularly limited, with the proviso that it is an enzyme having TGase activity. Examples of such an enzyme include those which are originated from mammals such as guinea pig and the like (cf. Japanese Patent Publication No. 1-50382), microorganisms such as of the genera Streptoverticillium and the like (for example, see Japanese Patent Application Kokai No. 64-27471) and fishes such as codfish and the like (for example, see Nobuo Seki et al., Japan Journal of Scientific Fisheries, vol.56, p.125, 1990) and those which are obtained by recombinant DNA techniques making use of biotechnology (cf. Japanese Patent Application Kokai Nos. 1-300,889, 5-199,883 and 6-225,775). Of these enzymes, microbial TGase is desirable from the view point that it can be produced in a large scale and does not require calcium for the expression of its enzyme activity.
Purity of the TGase to be stabilized also has no particular limitation. That is, either crude product or high purity product obtained by purification can be stabilized.
Next, a protein material to be used as a stabilizing agent is described. This is the specified substance described in the foregoing.
Examples of such a protein material broadly include plant proteins such as wheat protein, soybean protein and the like, animal proteins such as milk protein, the albumen, plasma protein, gelatin, cheese whey protein and the like and partial hydrolysates thereof, which have no particular limitation. Thus, it should be noted that partial protein hydrolysates are also included in the protein material of the present invention. Of these protein materials, the partial protein hydrolysate is particularly preferred, because it can be handled easily due to its small changes in viscosity when dissolved and it has high ability to keep the enzyme activity of TGas
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Hondo Keiko
Kuhara Chiho
Soeda Takahiko
Ajinomoto Co. Inc.
Lankford , Jr. Leon B.
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