JPH0578369A - Method for separating and recovering d-biotin - Google Patents

Abstract

PURPOSE:To separate and recover d-biotin in high yield by bringing an aqueous solution containing the d-biotin into contact with a nonionic exchange porous resin and eluting the d-biotin. CONSTITUTION:An aqueous solution containing d-biotin, e.g. a fermentation liquor or an enzymic reactional solution is brought into contact with a nonionic exchange porous resin at 5-60 deg.C. The d-biotin is then eluted with an eluent (e.g. ethanol-NH4OH) to afford the objective substance. The nonionic exchange porous resin is preferably a porous resin, having a giant network structure and a uniform pore diameter and comprising polystyrene or a polyacrylic ester as a resin parent substance and divinylbenzene as a cross-linking agent such as Diaion HP-10(R) (manufactured by Mitsubishi Kasei Corporation). The d-biotin is useful as medicines, foods, feed additives, etc.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for separating and recovering d-biotin from an aqueous solution containing d-biotin. According to the present invention, d-biotin can be separated and recovered in high yield.

[0002] d-Biotin is an industrially important compound as a medicine, food and feed additive.

[0003]

2. Description of the Related Art As a method for separating and recovering d-biotin, d-biotin
A method in which an aqueous solution (fermentation solution) containing biotin is brought into contact with activated carbon and then eluted with a suitable eluent (Japanese Patent Publication No. Sho 42).
-8918) has been proposed.

[0004]

However, in this production method, the recovery rate of d-biotin is extremely poor, and there is a problem to be improved as an industrial production method, and d-biotin can be recovered more efficiently. Development of a method was desired.

The inventors of the present invention have made earnest studies on the development of a highly efficient method for separating and recovering d-biotin, and by using a non-ion-exchangeable porous resin, a high-yield aqueous solution containing d-biotin can be obtained. The inventors have found that d-biotin can be separated from the above, and have completed the present invention.

[0006]

Means for Solving the Problems The present inventor has contacted an aqueous solution containing d-biotin (for example, a fermentation solution, an enzyme reaction solution) with a nonion-exchangeable porous resin, and then diluted ammonia water or the like. This is a method for separating and recovering d-biotin that is eluted with. According to this method, d-biotin can be efficiently separated and recovered.

The non-ion-exchangeable porous resin used in the present invention is preferably a porous resin having a huge network structure and a uniform pore size, in which polystyrene or polyacrylic ester having divinylbenzene as a crosslinking agent is used as a resin matrix. , Among them, for example, Diaion HP-10, the same H
P-20, HP-21, HP-30, HP-4
0, the same HP-50 (Mitsubishi Kasei), Amberlite XA
D-2, XAD-4, XAD-7, XAD-8
(ROHM, AND, House Co.), Levatit 00103
1 (Bayer) and the like can be preferably used.

The non-ion-exchangeable porous resin may be used under the general conditions for using the resin.
The liquid temperature at the time of carrying out the adsorption treatment with the non-ion exchangeable porous resin of the present invention is usually 5 to 60 ° C, preferably 10 to
The temperature is 40 ° C., and the pH at that time can be carried out over a wide range from strongly acidic to strong base, and there is no particular limitation.

Further, as the form of use of the resin for the treatment with the non-ion-exchangeable porous resin of the present invention, it may be suspended and stirred in the target liquid, or may be filled and passed through the column. The latter is more preferable in terms of efficiency. The flow rate at the time of filling and flowing the column is usually in the range of sv = 0.5 to 10, preferably 1 to 3, and the treatment liquid amount ratio to the resin amount is usually 1 to 100, preferably 5 to 5. The range is 20.

The non-ion-exchange porous resin after the treatment of the present invention is in contact with a dilute alkali, dilute acid or an organic solvent such as methanol or ethanol / water mixture such as water = 50/50. By doing so, it can be easily regenerated, can be repeatedly used, and is economically and industrially sufficiently practicable.

The present invention will be specifically described with reference to the following examples.

[0012]

【Example】

Example 1 500 ml of an aqueous solution containing 50 mg of d-biotin (HCl
PH adjusted to 2.5) with a column (H16) filled with 50 ml of non-ion exchange porous resin Diaion HP-20.
0 mm × φ30 mm) with sv = 1 and then distilled water 50
After washing with 0 ml of water, 50% ethanol / 28% NH ₄ OH
Elution was performed with 500 ml of an eluent of 18/1. Eluate 50
After concentrating 0 ml under reduced pressure, it was dried and the recovery amount was measured to find that d-biotin was 45 mg. As a comparative example, activated carbon (Diahope 008, manufactured by Mitsubishi Kasei Co., Ltd.) was used instead of the non-ion exchange porous resin under the same conditions, and the recovery amount of d-biotin was measured to be 31 mg.
Met. ..

Example 2 The same operation as in Example 1 was carried out except that the non-ion exchange porous resin shown in Table 1 was used. The results are shown in Table 1.

[0014]

[Table 1]

Example 3 A culture solution [(NH ₄ ) ₂ S containing 50 mg of d-biotin.
O ₄ 14g / l, KH ₂ PO ₄ 0.5g / l, K ₂ HP
_{O 4 0.5g / l, MgSO 4} · 7H 2 O 0.5g / l
, FeSO ₄・ 7H ₂ O 0.02g / l, MnSO ₄・
4-6 H ₂ O 0.02 g / l (adjusted to pH 7.6 with NH ₄ OH) 500 ml (adjusted to pH 2.5 with HCl) were used in the same manner as in Example 1 except that non-ion exchangeable porous material was used. As a result, the recovery amount of d-biotin was 44 mg, and the recovery amount of d-biotin was 28 mg in the comparative treatment with activated carbon.

Example 4 The same operation as in Example 3 was carried out except that the non-ion exchange porous resin shown in Table 2 was used. The results are shown in Table 2.

[0017]

[Table 2]

[0018]

INDUSTRIAL APPLICABILITY As described above, by using the non-ion-exchangeable porous resin, d-biotin can be efficiently separated and recovered from the d-biotin-containing aqueous solution.

Claims (1)

[Claims] 1. A method for separating and recovering d-biotin, which comprises contacting an aqueous solution containing d-biotin with a non-ion-exchange porous resin and then eluting with an eluent.