Polyamine-containing composition

Abstract

To provide: a polyamine-containing composition which is obtained from a bacterial body, a culture, or an extract of yeast as a raw material, the composition having a high spermidine content and a low spermine content; and a production method of the composition.SOLUTION: Purifying a yeast extract extracted from yeast with an ion-exchange resin can result in a polyamine-containing composition which includes a polyamine of 5 wt.% or more in terms of solid content and which has a high spermidine content and a spermine content at a particular ratio or less to the spermidine content.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a polyamine-containing composition obtained from yeast and a method for producing said composition. [Background technology]

[0002] Polyamines are one of the physiologically active substances present in large amounts in cells, and are known to have functions such as extending healthy lifespan (Non-Patent Document 1), suppressing inflammation (Non-Patent Document 2), enhancing intestinal barrier function, strong antimutagenicity, protection from oxygen stress, suppressing memory decline associated with aging by ingestion of a high-polyamine diet (Non-Patent Document 3), and promoting longevity by inducing autophagy by spermidine (Non-Patent Document 4). In particular, spermidine has been found to extend human healthy lifespan (Non-Patent Document 5). In addition, it has been reported in recent years that the activity of spermidine may be inhibited by spermine (Non-Patent Document 6).

[0003] To date, polyamine-containing compositions using yeast that can be cultured in large quantities at low cost have been reported, including polyamines prepared from the cells or culture medium of budding yeast (Patent Document 1), nutritional compositions containing yeast-derived nucleotides and polyamines (Patent Document 2), and spermidine-rich yeast and foods and beverages containing said yeast (Patent Document 3). In addition to yeast, polyamine-containing compositions obtained from microorganisms of the genus Bacillus, such as Bacillus subtilis var. natto, have also been reported (Patent Document 4).

[0004] However, since yeast contains not only spermidine but also spermine to a similar extent, the activity of spermidine may be inhibited by spermine in the polyamine-containing compositions disclosed in Patent Documents 1 and 2. In addition, since the spermidine-rich yeast of Patent Document 3 is produced by genetic recombination, it was an issue as to whether it would be accepted in the food market in particular. In addition, since it is generally known that microorganisms of the genus Bacillus form spores, the cost of cleaning the production equipment was a problem. For this reason, there was a demand for a polyamine-containing composition with a high spermidine content that can be produced at low cost without using genetically modified microorganisms. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 10-052291 [Patent Document 2] JP 2015-030688 A [Patent Document 3] WO2018 / 168525 Brochure [Patent Document 4] JP 2022-146649 A [Non-patent literature]

[0006] [Non-Patent Document 1] Sci Rep. 4:4548 (2014) [Non-Patent Document 2] Biomol. Ther. 21:1 (2013) [Non-Patent Document 3] Nat. Neurosci. 16:1453 (2013) [Non-Patent Document 4] Nat. Cell Biol. 11:1305 (2009) [Non-Patent Document 5] Alzheimers Res. Ther. 11:36 (2019). [Non-Patent Document 6] Science. 378:eabj3510 (2022) Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a polyamine-containing composition having a high spermidine content and a low spermine content, which is made from the fungus bodies or culture broth of Torula yeast as a raw material, and a method for producing said composition. [Means for solving the problem]

[0008] As a result of intensive research into solving the above-mentioned problems, the present inventors have found that a polyamine-containing composition having a high spermidine content and a low spermine content can be easily obtained by purifying a yeast extract extracted from yeast cells with a resin.

[0009] That is, the present invention relates to the following first to fifth aspects. First, the polyamine-containing composition contains polyamines in an amount of 5% by weight or more calculated as solid content, and spermidine in all polyamines accounts for 60% by weight or more. A second aspect is the polyamine-containing composition according to the first aspect, in which the content of spermine is 0.45 parts by weight or less per part by weight of spermidine. Third, the polyamine-containing composition according to the first or second aspect of the present invention is a polyamine-containing composition in which the content of 5'-guanylic acid in the composition is 5% by weight or less in terms of solid content as disodium heptahydrate. Fourthly, there is provided a method for producing the polyamine-containing composition according to either the first or second aspect, which comprises a step of passing a yeast extract through a cation exchange resin. Fifth, there is provided a method for producing the polyamine-containing composition according to the third aspect, which comprises a step of passing a yeast extract through a cation exchange resin. Effect of the Invention

[0010] According to the present invention, by using yeast as a raw material, it is possible to obtain a polyamine-containing composition having a polyamine content of 5% by weight or more calculated as solid matter and a spermidine content in the polyamines of 60% by weight. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The polyamine in the polyamine-containing composition of the present invention refers to putrescine, spermidine, spermine, cadaverine, and agmatine, which is a precursor of polyamine. The polyamine-containing composition of the present invention may contain two or more kinds of the polyamine. The polyamine-containing composition of the present invention preferably contains 5% by weight or more of the polyamine in terms of solid content, more preferably 10% by weight or more.

[0012] The amount of spermidine is preferably 60% by weight or more, more preferably 65% ​​by weight or more, based on the total amount of polyamines in the composition. The amount of spermine is preferably 0.45 parts by weight or less, based on 1 part by weight of spermidine in the composition. Furthermore, the content of 5'-guanylic acid contained in the composition is preferably 5% by weight or less, more preferably 3.5% by weight or less, calculated as solid content as disodium heptahydrate.

[0013] The polyamine-containing composition of the present invention can be obtained from a yeast extract. As long as the composition of the present invention can be obtained, there is no limitation on the type of yeast used as the raw material for the yeast extract, and budding yeast (Saccharomyces cerevisiae) or torula yeast (Siblindonella jadini; also known as Candida utilis) can be used, but torula yeast is more preferred. For example, the torula yeast cells cultured under general conditions are subjected to an extraction operation such as hot water extraction, and the extract obtained is treated with nuclease or transaminase, then centrifuged and the supernatant is removed. The remaining fraction is passed through a cation exchange resin to adsorb polyamines to the resin, and then eluted with a basic aqueous solvent of about 1 to 5%, thereby obtaining the polyamine-containing composition of the present invention. In addition, a commonly available yeast extract derived from torula yeast may be used as the fraction passed through the cation exchange resin, or a commonly available nuclease preparation or transaminase preparation may be applied to the yeast extract, centrifuged, and the remaining fraction removed from the supernatant may be used. The cation exchange resin used is preferably a cation exchange resin having a strongly acidic ion exchange group such as a sulfonic acid group in the structure. The treatment conditions such as the liquid passing speed are not particularly limited as long as the polyamine can be adsorbed. In addition, the basic aqueous solvent used for elution can generally be a basic aqueous solvent having ion exchange and regenerative properties for the cation exchange resin, for example, an aqueous sodium hydroxide solution or an aqueous potassium hydroxide solution, and an aqueous sodium hydroxide solution is preferred.

[0014] The composition obtained by the above method can be used as the polyamine-containing composition of the present invention, and if necessary, steps such as desalting and ultrafiltration can be added. In addition, the composition may be subjected to steps such as neutralization, concentration, addition of excipients, drying, etc., which are generally used in food production.

[0015] The polyamine-containing composition of the present invention can be taken orally as a supplement or the like, but can also be added to food and drink. The form of the supplement is not particularly limited, and can be in the form of tablets, capsules, soft capsules, or nutritional drinks. In addition, it can be used as a mixture with other materials as long as it does not inhibit the effect of the present invention.

[0016] The polyamine-containing composition of the present invention can be used as it is as a cosmetic composition, a pharmaceutical composition, or a topical composition. The form of the cosmetic composition, pharmaceutical composition, or topical composition is not particularly limited, and it can be prepared in the form of a solution, dispersion, paste, gel, or sheet according to a conventional method using a solvent or base material generally used in cosmetic compositions, pharmaceutical compositions, and topical compositions. It may be used as a mixture with other materials as long as it does not inhibit the effects of the present invention.

[0017] The content of polyamine contained in the composition of the present invention can be measured by a general analytical method. For example, it can be quantified by liquid chromatography-quadrupole mass spectrometry using various free polyamines with known concentrations as standards.

[0018] Furthermore, the amount of 5'-guanylic acid contained in the composition of the present invention can be quantified by a general analytical method using high performance liquid chromatography, and the content as disodium heptahydrate can be quantified.

[0019] Since the composition of the present invention contains a predetermined amount or more of spermidine, it can be taken orally as a supplement or can be added to food and drink. It can also be used as a cosmetic composition, a pharmaceutical composition, a composition for external use, etc. Furthermore, it may be prepared in a desired form as described above and used. Since the composition of the present invention has a spermine content relative to spermidine of a predetermined amount or less, it is considered that the inhibition of the physiological activity of spermidine by spermine is suppressed. EXAMPLES

[0020] The present invention will be specifically described below using examples, but the present invention is not limited to these.

[0021] <Polyamine Analysis Method> Using high performance liquid chromatography-single quadrupole mass spectrometry (HPLC-MS), the polyamines in the sample are measured as the total amount, using the free polyamines spermidine and spermine as standards. HPLC conditions Separation column: Scherzo SM-C18 (37°C, 3.0 x 150 mm, Imtakt) Mobile phase: A: 30 mM ammonium acetate and acetic acid (100:0.2), B: water, acetonitrile, and acetic acid (80:20:0.8) ·Flow rate: 0.4 mL / min Gradient: 0 → 3.5 min, 0 → 20% solution B, 3.5 → 10 min, 20 → 100% solution B. Detection: m / z 89.1 (putrescine), m / z 146.2 (spermidine), m / z 203.3 (spermine)

[0022] <Preparation of polyamine-containing composition> A commercially available yeast extract was used as the yeast extract derived from Torula yeast. 100 g of this yeast extract contained 200 mg of spermidine, 98 mg of spermine, 65 mg of putrescine, and 10.5 g of disodium guanylate heptahydrate. This yeast extract was dissolved in water to a concentration of 5 w / v%, and a commercially available nuclease preparation and transaminase preparation were added to the resulting aqueous solution at 0.1 wt% each, and the mixture was reacted at pH 5.2 and 45°C for 16 hours. The supernatant was then removed by centrifugation, and the pH was adjusted to 3.0 by adding sulfuric acid. The mixture was then extracted by heating at 55°C for 1 hour, and then sterilized and inactivated at 95°C for 30 minutes. The supernatant obtained by centrifuging the extract was passed through a cation exchange resin having sulfonic acid groups as ion exchange groups at 150 BV to adsorb polyamines. The resin was then thoroughly washed with ion exchange water, and 20 BV of a 2% aqueous sodium hydroxide solution was passed through it to recover the adsorbed polyamines.

[0023] <Polyamine analysis results> The obtained polyamine-containing composition was quantitatively analyzed by HPLC-MS. The results were that the solid content was 13.0% spermidine, 2.8% spermine, and 0.6% putrescine. The spermine content was 0.22 parts by weight per part by weight of spermidine. The 5'-guanylic acid content was 1.1% by weight in terms of solid content as disodium heptahydrate.

[0024] From the above, it was shown that a polyamine-containing composition containing 5% by weight or more of polyamine in terms of solid content and 60% by weight or more of spermidine in the total polyamine can be easily obtained. In addition, since the obtained composition has a low spermine content relative to spermidine, it can be used orally or externally as a composition in which the inhibition of spermidine activity by spermine is suppressed.

Claims

1. A polyamine-containing composition, in which the amount of polyamine in the composition is 5% by weight or more calculated as solid content, and spermidine accounts for 60% by weight or more of all polyamines.

2. 2. The polyamine-containing composition according to claim 1, wherein the polyamine-containing composition contains 0.45 parts by weight or less of spermine per 1 part by weight of spermidine.

3. 3. The polyamine-containing composition according to claim 1, wherein the content of 5'-guanylic acid in the composition is 5% by weight or less in terms of solid content as disodium heptahydrate.

4. A method for producing the polyamine-containing composition according to claim 1 or 2, comprising a step of passing a yeast extract through a cation exchange resin.

5. A method for producing the polyamine-containing composition according to claim 3, comprising a step of passing a yeast extract through a cation exchange resin.