Aqueous L-lysine-containing animal feed supplements and...

Food or edible material: processes – compositions – and products – Treatment of live animal

Reexamination Certificate

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C426S061000, C426S656000, C426S807000

Reexamination Certificate

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06340486

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates to aqueous L-lysine-containing animal feed supplements, which optionally contain preferably inactivated microorganisms from the fermentation process, and a process for the production of such supplements.
2. Background Information
The essential amino acid L-lysine is widely used as an animal feed supplement.
It is known that L-lysine is produced by fermentation of strains of coryneform bacteria, in particular Corynebacterium glutamicum. Due to its great significance, efforts are constantly being made to improve the production process. Improvements to the process may relate to the intrinsic performance characteristics of the microorganisms by for example mutagenesis and selection, to measures concerning fermentation technology, for example stirring and oxygen supply, to the composition of the nutrient media, such as for example sugar concentration during fermentation, or to working up to the product presentation by, for example, ion exchange chromatography or crystallization.
According to the prior art, there are three different groups of L-lysine-containing product presentations which are produced from L-lysine-containing fermentation broths. The best known group comprises pulverulent or crystalline product presentations obtained from cleaned-up or pure L-lysine, which is typically present in the form of a salt such as, for example, L-lysine monohydrochloride. Another group of solid product presentations, such as are described in EP-B-0533039, for example, contains, in addition to L-lysine, the feedstock used during fermentative production and optionally the inactivated biomass of the microorganism used.
Pulverulent, crystalline or indeed granular product presentations exhibit clear disadvantages with regard to handling properties and application. The quality of the product may be influenced to a considerable extent by the weather, for example, high air humidity levels, since the caking which may occur as a consequence of such atmospheric influences impairs the shelf life and dispensing properties of the product when used. Processing of dry, pulverulent L-lysine-containing products may lead, in turn to undesirable dust development.
The third group of L-lysine-containing products comprises concentrated, aqueous, alkaline solutions and thereby circumvents the above-mentioned problems (EP-B-0534865). Given the various product presentations, many different processes are known for producing L-lysine-containing products from L-lysine-containing fermentation broths. There are essentially two different processes currently known for producing solid, pure L-lysine.
Lysine may be obtained as a monohydrochloride (Lys-HCl) by crystallization from the corresponding fermentation broths, after the inactivated biomass has been separated off by suitable methods. Cleaning-up of the filtrate prior to further concentration is performed in general by ion exchange chromatography over several stages. First of all, the fermentation broth separated from the biomass is acidified, preferably by the addition of hydrochloric acid (HCl) or sulfuric acid (H
2
SO
4
), to ease adsorption of the lysine on the ion-exchange resins. In addition to the L-lysine produced by fermentation, various other cations which are present in the fermentation broth are also bound. In general, various ion-exchange columns connected in sequence are necessary for obtaining a pure product. The adsorbed lysine is then preferably eluted by an ammoniacal solution and the ion-exchange column is regenerated. The lysine solution obtained in this way is then concentrated and lysine-HCl is obtained in crystalline form after neutralization with hydrochloric acid.
Another method enables lysine to be obtained in the form of a crystalline salt after purifying with activated carbon (SU-183581). The lysine-containing fermentation broth is inactivated by standard processes using moist heat and separated off from the biomass by filtration. After acidification of the filtrate to pH 5, 4-5% activated carbon is added with constant stirring at 50-55° C., in order to separate off undesirable impurities from the filtrate and to prevent discoloration of the crystallizate. In a further filtration stage thereafter, the activated carbon is separated off and the dissolved sulfate is then precipitated as calcium sulfate by the addition of calcium hydroxide. This is filtered off, the ammonia content being removed in a rotary evaporator under vacuum and the solution being concentrated until crystallization occurs on cooling.
The disadvantage of these two preparation methods lies in the numerous individual stages and the complex cleaning processes using ion-exchange chromatography. The elimination of troublesome salts or the use of different elution media creates additional waste streams, which have either to be cleaned by complex methods or expensively disposed of.
EP-B-0533039 counters these disadvantages in that all the fermentation feedstock, optionally including the biomass, is concentrated and spray-dried at elevated temperature. U.S. Pat. No. 5,990,350 describes a process for producing biomass-free L-lysine-containing granules. In this process, the biomass is first isolated from the fermentation broth by ultrafiltration and discarded. The filtrate recovered in this way is then concentrated by extensive evaporation of the water content. The concentrate obtained in this way is finally dried and worked up in a complex manner by fluidised bed spray granulation to produce granules. A process is known from EP-B-0534865 for producing aqueous, basic, L-lysine-containing solutions from fermentation broths. In the process described therein, the biomass is separated from the fermentation broth and discarded. A pH value of between 9 and 11 is established using a base such as sodium, potassium or ammonium hydroxide, for example. The mineral constituents (inorganic salts) are separated from the broth by crystallization after concentration and cooling and either used as fertiliser or discarded (land-filled).
SUMMARY OF THE INVENTION
Object of the Invention
The object of the invention consists in providing novel, aqueous preparations of L-lysine and its salts suitable as a feed supplement. In addition, it is an object of the invention to provide a production process that is more economical and efficient than processes currently known.
Description of the Invention
The essential amino acid lysine is widely used as an animal feed supplement.
Microorganisms of the genus Corynebacterium are distinguished by the capacity to secrete high concentrations of L-lysine into the corresponding fermentation medium over a relatively short period. The production processes are generally performed as fed batch processes. L-lysine produced by fermentation is currently predominantly worked up to produce a crystalline product, powder or granules.
The present invention provides a fermentation broth-based animal feed supplement, characterized in that it contains
a) L-lysine and/or salts thereof,
b) the biomass formed during fermentation in an amount of from 0 to 100%, preferably from 50 to 100% and very particularly from 90 to 100%,
c) at least the majority of the other dissolved and suspended constituents of the fermentation broth are present in liquid form, optionally as a suspension and
e) has a pH of less than 7.
The animal feed supplement has a lysine content (as lysine base) of 10 wt. % to 35 wt. %, preferably 15 wt. % to 35 wt. %, particularly preferably 18 wt. % to 35 wt. % and very particularly preferably 21 wt. % to 34 wt. %, relative to the total amount of supplement. The pH is less than 7, preferably 2 to 6.5, particularly preferably 2.5 to 6 and very particularly preferably 2.5 to 5.
The total solids content of the supplement amounts to from 10 wt. % to 55 wt. %, preferably 20 wt. % to 55 wt. %, particularly preferably 35 wt. % to 55 wt. % and very particularly preferably 45 wt. % to 54 wt. %. The solids content contains, in addition to lysine
preferably the biomass of the producing microorganism,
the inorga

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