GB1563954A - Preparation of vitamin - Google Patents

Description

(54) IMPROVEMENTS RELATING TO THE PREPARATION OF VITAMIN B, (71) We, VYSOKA SKOLA CHEMICKO TECHNOLOGICKA, of Praha, Czechoslovakia, a corporation organised and existing under the laws of the Czechoslovak Socialist Republic, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method bv which it is to be performed, to be particularly described in and by the following statement: The present invention relates to a method of producing vitamin B, in a fermentation process, either independently or during the production of fermented beverages.

The recent human nutrition mode consisting in a predominant consumption of white finely ground flour type and heat preserved foods leads to a suboptimum con centration of vitamin B, in the foodstuff and to an increased demand for synthetic vitamin B1 for pharmaceutical purposes, or for fortification of some flour types as well as products therefrom. In the production of a majority of alcoholic beverages, for cxample, beer, wine, fruit wines, cider, sake, sorghum, quass such yeast strains are usually employed which deprive the fermenting medium of vitamin B, the source of which is various raw materials, for example malt, cereals and fruits. The vitamin becomes part of cells which, however, arc removed from fermenting media after the fermentation. Thus for instance, with beer the essential raw material of which, viz.

malt, is very rich in vitamin Bi, a decrease in vitamin B, content of up to 3 Jig per litre occurs, due to the action of brewer 5 yeast.

The recovery of vitamin B, from brewer's or wine yeast waste however, is not sufficient to cover the demand for this vitamin for rational population nutrition and for pharmaceutical purposes.

Chemical synthesis of thiamine is a relatively elaborate technological process accompanied moreover bv problems of waste water treatment. Vitamin B, has not previously been produced in a one-stage fer mentation process since no microorganisms have previously been knawn, which would produce this vitamin an economical scale from simple carbon sources, for example sugars.

The disadvantages of the prior art as hereinbefore referred to are substantially reduced or eliminated by a method of producing vitamin B1 in a fermentation process according to the present invention by means of mutants of yeasts of genus Sac charol,lyces Meyen emencl. Reess. We have prepared such mutants by genetic and biochemical methods. The mutants are able to be repeatedly used for the production of vitamin B, by fermentation. These mutants are able lo synthesize vitmin Bi in a high degree even if the vitamin is present in the fermentation medium and to excrete the vitamin. Mutants of Saccharomyces cerevisiae Hansen designed as DBM 159 - and of Saccharomyces uvarum Beijerinck (syn.

Saccch. carlsbergeiisis Hansen), designated as DBM 189 have proved to be particularly suitable for this purpose.

According to the invention. in a method of preparing vitamin B1 at least one mutant of yeast of the genus Saccharomyces Me yen emend, Reess is used to synthesise said vitamin from a fermentation medium comprising suitable sources of carbon and nitrogen and from mineral salts including a source of sulphur, and the vitamin so produced is excreted into the fermentation medium from which it may be isolated.

The mutant of yeast is preferably a mut ant of Saceharomyces cerevisiae Hanker, strain DBM l.Se, or Saccharoinyca ii van tin Beijerinck (syn. Saccharomyces carlsbergen- si.s Hansen) strain DBM 189.

The carbon source desirably comprises a source of sugar such as molasses, starch syrup, or cellulose hydrolysate. The nitrogen source may comprise for example ammonium sulphate.

The fermentation medium may also include a phosphorus source e.g. ammonium phosphate, and small quantities of metal ions, for instance magnesium (II), potas- sium (I) ions. Vitamin B1 can be obtained from the fermentation medium e.g. by passing the completely fermented liquor freed of yeast through an ion exchange column and by isolating the vitamin from the column eluate. In the production of alcoholic beverages, for instance, beer, wine, fruit wines, cider, sake, sorghum quass, and lactic fermentation beverages, for example kefir, kumis, the above-mentioned yeast strains are used as the inoculum or added to the inoculum so that the vitamin which is excreted by the yeast during the fermentation process remains a part of the beverage and fortifies it.

In order that the invention be fully understood some preferred embodiments thereof will now be described with reference to the following Examples which, however, are not intended to limit in any way-the scope of the inventlon.

EXAMPLE 1 Saccharomyces cerevisiae Hansen, strain DBM 159 was propagated from a laboratory culture in a conventional manner, e.g. by successive propagation of the culture from malt extract. The fermentation of molasses mash, acidified to pH 4.2 to 4.5, and with added ammonium sulphate and ammonium phosphate took place at a saccharization temperature of 220C and a temperature of 28 to 300C of the completely fermented liquor. From the fermented liquor from which yeast had been removed, preferably by centrifuging in a continuous centrifuge, the vitamin Bl was obtained, for example, by passing it through an ion exchange column. Thereafter the liquor was processed in a manner conventional in the distillery practice. The vitamin was eluted from the ion exchange column and isolated from the eluate in a conventional manner.

for instance, by precipitating it with silver nitrate and decomposing the precepitate with dilute hydrochloric acid. The vitamin B1 yield was about 10 mg per litre of completely fermented liquor, the ethnol yield being practically unaffected.

EXAMPLE 2 Sacchoromyces cerevisiae Hansen strain DBM 159, or Saccharomyces uvarum Bei- jerinck (sell S. caribergensis Hansen) strain DBM 189 was propagated in a conventional manner. The fermentation of wort took place at a temperature of from 6 to 100C, After the yeast had deposited the fermented wort was skimmed. and beer was aged 2 to 4 weeks at 40C and further processed in a conventional manner. The yeast taken from the main fermentation process could be reused up to 6 times. The beer prepared in this way hy means of the aforementioned yeast contained, on the one hand, the vitamin B, from the raw materials employed and, on the other hand, the vitamin produced by the Succharomyces strain used.

WHAT WE CLAIM IS:- 1. A method of preparing vitamin Bl, wherein at least one mutant of yeast of the genus Sacchara,nyces Me yen emeltd. Reess is used to synthesise said vitamin from a fermentation medium comprising suitable sources of carbon and nitrogen and from mineral salts including a source of sulphur, and the vitamin so produced is excreted into the fermentation medium from which it may be isolated.

2. A method according to Claim 1, wherein said mutant of yeast is a mutant of Saccharornyces cerevisiae Manse), strain DBM 159, or Saccharomyces uvarrmz Bei jerinek (Syn. Saceharomyces carlsbergensis Hansen) strain DBM 189.

3. A method according to Claim 1 or Claim 2, wherein said carbon source comprises a source of sugar.

4, A method according to Claim 3, wherein said source of sugar comprises molasses, starch syrup or cellulose hydrolysate.

5. A method according to any one of the preceding claims, wherein said nitrogen source comprises ammonium sulphate.

6. A method according to any one of the preceding claims, wherein said fermentation medium further comprises a source of phosphorus.

7. A method according to any one of the preceding claims. wherein said fermentation medium further comprises small quantities of metal ions.

8. A method according to any one of the preceding claim, wherein a quantity of said vitamin is originally present in the fermenting medium.

9. A method according to any one of the preceding claims, wherein said vitamin B1 is produced as a by-product during ethanol manufacture.

10. A method according to any one of the preceding claims further comprising isolating said vitamin from said fermentation medium.

11. A method according to any one of Claims I to 9, wherein said at least one mutant for producing vitamin B1 serves as a starting material or is added to a starting material for the manufacture of a fermented beverage.

12. A method according to Claim 11, wherein said beverage is beer, wine, fruit wine, cider, quass, sorghum, cake, kefir or kumis.

13, A method of preparing vitamin B1, the method being substantially as hereinbefore described with reference to Example 1 or Example 2.

I4. Vitamin B1 when prepared by a method according to any one of the pre

Claims (1)

1. ceding claims.

   15. A process for the production of a pharmaceutical product, including a method according to any one of Claims 1 to 10, or Claim 13 (with reference to Example 1 only) of preparing vitamin BI.

   16. A process for the production of a beverage, including a method according to iany one of Claims 1 to 9 or any one of Claims 11 to 13.

   17. A pharmaceutical product when produced by a process according to Claim 15.

   18. A beverage when produced by a process according to Claim 16.