Diacetyl production

Abstract: A process for the preparation of diacetyl which comprises fermenting an aqueous pectin slurry with a lactic acid producing bacteria at a temperature from 16°C to 38°C, a pH of 4.0 to 7.0 for a period of from 8 to 24 hours under aerobic conditions.

Description: The present invention relates to a process for the production of diacetyl. Diacetyl is a major constituent of butter flavour, ordinary butter containing about 8-10 ppm. It is also an important flavour compound in dairy products and coffee. Up to the present time, milk has been used as a source of diacetyl by fermentation which yields about 80 ppm. USP 4678673 describes the preparation of diacetyl from oilseed products by fermenting with Lactobacillus casei spp rhamnosus AT CC 39595 at a pH of 6 to 7 for less than 8 hours but the yield of diacetyl is never more than 44 ppm. USP 4867992 describes the preparation of a buttery flavour containing diacetyl by fermenting a coffee substrate with a lactic acid producing bacteria. EP-A-0 430 406 describes a method for the fermentative production of diacetyl and acetoin, which comprises culturing a lactic acid bacterium in a culture medium containing a sugar source which said lactic acid bacterium can utilize (glucose, lactose, etc.), in the presence of at least one additive selected from iron porphyrin and the like, and also in the presence of at least one metal salt. We have found surprisingly that by fermenting a pectin substrate with a strain of lactic acid producing bacteria, we can obtain yields of diacetyl of above 500 ppm. Accordingly, the present invention provides a process for the preparation of diacetyl which comprises fermenting an aqueous pectin slurry with a lactic acid producing bacteria at a temperature from 16°C to 38°C and a pH of 4.0 to 7.0 for a period of 8 to 24 hours under aerobic conditions. The fermentation media may be prepared by adding sodium chloride to the pectin slurry, and the alkaline salts of pyruvic and citric acid preferably the sodium salts. The amount of sodium chloride may be from 0.25 to 2.5%, and preferably from 0.5 to 1.5% by weight based on the weight of the slurry. The amount of pyruvate may be from 0.25 to 2.5% and preferably from 0.5 to 1.25% by weight based on the weight of the slurry. The amount of citrate may be from 0.5 to 5% and preferably from 1.5 to 3.5% by weight based on the weight of the slurry. The fermentation media may then conveniently be autoclaved, for example from 100 -120°C over 5 to 30 minutes. The lactic acid producing bacteria may be one known to be capable of producing diacetyl e.g. Leuconostoc cremonis, Streptococcus lactis subspecies diacetylactis, but conveniently, the lactic acid producing bacteria is Lactobacillus casei (hereinafter referred to as L.casei). Examples of suitable strains are L.casei ATCC (American Type Culture Collection) 334 isolated from Emmental cheese, L.casei ATCC 11582 - lactose negative and L.casei subspecies tolerans ATCC 25599 isolated from milk. The concentration range of L. casei is conveniently from 5 x 10 5 to 1 x 10 9 cells and preferably from 5 x 10 6 to 1 x 10 8 cells per millilitre of the slurry. The fermentation temperature is preferably from 30° to 35°C. The pH during the fermentation is preferably from 5 to 5.75. The duration of the fermentation is preferably from 12 to 20 hours. The aeration may be achieved by bubling in air or oxygen and agitating, conveniently with stirring at 100 to 300 RPM, preferably from 225 to 275 RPM. The pectin in the slurry may be either apple pectin or citrus pectin or a mixture of pectins. The pectin slurry is preferably prepared from distilled water, or deionised water. The concentration of the pectin in the slurry may be from 5 to 50%, preferably from 10 to 40% and especially from 15 to 30% by weight. Advantageously, before fermentation the pectin substrate is incubated with pectinase. The amount of pectinase may be from 0.1 to 10% preferably from 0.2 to 5% and especially from 0.5 to 2.5% by weight based on the weight of the pectin slurry. The incubation temperature may be from 35° to 55°C and preferably from 40° to 50°C. The duration of the incubation may be from 30 minutes to 6 hours, preferably from 1 to 3 hours. Advantageously, the incubation mixture is agitated, for example by stirring at 100 to 500 RPM, preferably from 200 to 300 RPM. After incubation with pectinase, large precipitates are preferably removed, for example, by filtration. Afterwards, the pectinase treated mixture may be autoclaved, for example, from 100-120°C over 5 to 30 minutes, or it may be used directly to prepare the media for fermentation with the lactic acid producing bacteria as hereinbefore described followed by autoclaving. The following example further illustrates the present invention. Parts and percentages indicated are by weight. Example 10 parts of apple pectin and 10 parts of lemon pectin together with 80 parts of distilled deionised water were treated with 1 part of pectinase by incubating at 40°C for 2 hours with stirring at 250 RPM. To the above pectinase treated pectin bean slurry were added 0.85% NaCl, 0.75% sodium pyruvate and 2.5% sodium citrate and the final media autoclaved at 110°C for 15 minutes. L. casei ATCC = 334 was added at a concentration range of 1 x 10 7 cells per millilitre of the slurry and fermentation took place at 32°C; pH 5.5 over 18 hours with stirring at 250 RPM. The fermented slurry was pasteurised and the resultant product was found to contain 1700 ppm of diacetyl.

Claims: A process for the preparation of diacetyl which comprises fermenting an aqueous pectin slurry with a lactic acid producing bacteria at a temperature from 16°C to 38°C, a pH of 4.0 to 7.0 for a period of from 8 to 24 hours under aerobic conditions. A process according to claim 1 in which the fermentation media contains sodium chloride, sodium citrate and sodium pyruvate. A process according to claim 1 wherein the pectin slurry contains apple pectin or citrus pectin. A process according to claim 1 wherein, before fermentation, the pectin slurry is incubated with pectinase. A process according to claim 1 wherein the lactic acid producing bacteria is L.casei.

Inventors: DUBOFF SHIRLEY ANN, KWON STEVEN SOON-YOUNG, VADEHRA DHARAM VIR, DUBOFF, SHIRLEY ANN, KWON, STEVEN SOON-YOUNG, VADEHRA, DHARAM VIR
Applicants: NESTLE SA, SOCIETE DES PRODUITS NESTLE S.A.
Application year: 1993
Main classification: C12P7
IPC codes: C12P7, C12R1, C12P19, A23L1