MXPA06010474A - Method for enriching trehalose with the aid of alumosilicates - Google Patents

Abstract

The invention relates to a method for enriching trehalose from solutions, wherein the enrichment is performed with the aid of an adsorbent. The invention is characterized in that the adsorbent is an alumosilicate. The alumosilicate is preferably a zeolite. The invention also relates to the enrichment and purification of trehalose from fermentation broths, more particularly as coupled product from the fermentative production of other value products.

Description

METHOD FOR ENRICHING TREHALÓSA WITH THE ALUMINOSILICATOS AID The invention is further related to a process for enriching trehalose from solutions, in which trehalose is enriched using an adsorbent. The disaccharide trehalose (α-D-glucopyranosyl-α-D-glucopyranoside) consists of two glucose molecules which are conveniently linked together through a bond of a, a-1, l. Trehalose, due to its properties which are of interest in terms of yield is of increasing importance for the industry. An important field of application is to stabilize proteins and peptides, for example, enzymes and vaccines. A preferred use for trehalose is in the food industry. Trehalose is also used as a substitute for sucrose because of its reduced sweetness and its properties which preserve flavor. In addition, trehalose has a stabilizing action in freezing and drying operations. An additional field of application is in the cosmetics sector. Trehalose is preferably produced enzymatically or by fermentation using suitable microorganisms (Schiraldi, C, et al (2002), Trehalose Production: Exploiting Novel Approaches, Trends in Biotechnology, vol.20 (10), pages 420-425). Frequently, trehalose is also formed as a by-product in fermentations which serve for the production of other substances (Hull, SR, Gray, JSS, et al. (1995).) Trehalose as a Common Industrial Fermentation Byproducts. 266, pages 147-152). In particular, in the case of fermentations, other than chemical synthesis, highly contaminated solutions are formed, which may contain, for example, cells, proteins, lipids or other sugars. Trehalose should also be enriched from such highly contaminated solutions, and, depending on the intended use, be further purified. In the prior art, several enrichment and purification processes for trehalose are known. US 5,759,610 discloses a process for purifying trehalose from cultures of microorganisms comprising the steps of filtration and centrifugation, treatment with activated charcoal, deionization, purification with ion exchangers, concentration to form syrup products, further purification by techniques of column chromatography such as ion exchange column chromatography, activated carbon chromatography and silica gel column chromatography and also precipitation with organic solvents such as alcohol and acetone and filtration through membranes, and yeast fermentation or treatment alkaline in order to remove or disintegrate any remaining saccharides. For further purification, crystallization is proposed by cooling or spray drying, for example. The adsorption of trehalose to an adsorbent is not carried out JP 07000190 (Tradashi, E., et al.) Describes the isolation of trehalose from solid residues of beer fermentations. The residue is extracted with alcohol and / or treated with ultrasound to extract the trehalose from the residue. In addition, the trehalose enzyme present in the residue is inactivated by thermal treatment. Purification is performed, inter alia, through ion exchange columns and an activated carbon column. Trehalose does not adsorb to the columns in this process. U.S. Patent 5,441,644 describes a process in which trehalose is described from a fermentation broth. In the process, inter alia, ultrafiltration and decolorization are carried out using activated carbon. Trehalose does not adsorb to activated carbon in the process. A disadvantage of such processes appears to be that the respective adsorbents are used only for the adsorption of unwanted foreign matter, but do not adsorb the trehalose itself. Since the extraction and purification stages must adapt to the different foreign matter, these are complicated and only apply with difficulty on an industrial scale. In particular, this applies to purification from fermentation broths in which the trehalose content is usually less than 15% dry weight (Schiraldi et al. (2002), Trehalose Production: Exploiting Novel Approaches.) Trends in Biotechnology , vol 20 (10), page 421). According to another process, trehalose was purified as a byproduct of a fermentation by sequential chromatography on activated carbon and Bio-Gel P-2 (Hull, SR, Gray, JSS, et al., 1995.) Trehalose as a Byproduct of Common Industrial Fermentation, Carbohydrate Research, vol.226, pages 147-152). The process, however, is only a detection method, not a process which is suitable for an industrial scale. US 5,441,644 mentions, in addition to the process described above, a further process of the prior art in which a solution of acetonitrile containing trehalose is subjected to chromatography on silica gel. The publication mentions that these chromatographic processes are undue, however, for enrichment of trehalose or purification of trehalose on an industrial scale. Buttersack et al. (Specific Adsorption from Aqueous Phase on Apolar Zeolites, Progress in Zeolite and Microporous Materials, vol 105, pp. 1723-1730, 1997) describes the binding of certain mono and disaccharides to selected FAU, PEA and MFI zeolites. For individual disaccharides, highly divergent adsorption properties were found. Trehalose was not studied. In an additional work, Buttersack et al., Describe the binding of disaccharides to divergent Y zeolites and dealuminated Y zeolites (Buttersack et al., 1994) Adsorption of Glucose and Fructose containing Disaccharides on Different Faujasites Studies in Surface Science and Catalysis, vol 84, pp. 1363-1371). These emphasize the importance of the fructose radical in the disaccharides studied for the adsorption to zeolites. Trehalose was not studied and does not have a fructose radical either. A disadvantage of the previous adsorbents is that they have very general adsorption properties and can not be individually adjusted for the respective process. Therefore, there is a requirement for processes to enrich trehalose from solutions that use better absorbers, in particular for absorbers which can be designed to the respective process. It is an object of the present invention, therefore, to provide such a process, in particular for use in chromatographic processes. It is a further object of the present invention to provide a process which makes it possible to enrich trehalose from the fermentation broths, in particular from the lysine production fermentation broths. It has been found that this object is achieved from the known process to enrich trehalose from solutions using an adsorbent. A feature of the inventive process is that the adsorbent is an aluminosilicate. Compared with the adsorbents used according to the prior art (for example, activated carbons and ion exchangers), aluminosilicates, in particular zeolites, offer the advantage that a large number of variants can be prepared, and as a result the adsorbent can be better adapted to the problem of separation. Trehalose can be produced by a multiplicity of known processes. Traditionally, trehalose is produced by fermentation processes, with, meanwhile, enzymatic production processes that have come to be established (Schiraldi, C, et al., (2002) Trehalose Production: Exploiting Novel Approaches, Trend in Biotechnology, vol. 10), pp. 420-425). In microorganisms, 3 main enzymatic pathways for trehalose synthesis have been discovered; (1) a system of phosphorylase in fungi and yeasts, (2) a glycosyltransferase-hydrolase system in mesophilic and extremophilic bacteria and (3) a transglicosylation catalysed by trehalose synthase from maltose to trehalose (eg JP 09098779, KR99029104). The term enrichment is known to those skilled in the art. According to the present invention, the term enrichment is related in particular to the increase in the ratio of trehalose to undesired foreign matter. Normally, this ratio of trehalose corresponds to the dry weight of the product. In the preferred embodiment, the term enrichment is also related to the purification of trehalose. The term purification is known to those skilled in the art. In the present context, this is in particular a purification purpose to achieve a purity of trehalose in which trehalose is essentially free of other substances. In particular, this means trehalose in crystalline form. An enrichment or purification process is only economically convenient if the field is satisfactory. Therefore, it is an additional purpose of the present process to achieve not only a high enrichment but also a high yield. With respect to the solution, there are no special restrictions with respect to solvents, which can be used, for example, water or acetonitrile. Preferably, the solution is an aqueous solution.

An adsorbent within the meaning of the present invention is a solid or a gel-like substance on the surface in which the adsorption of another substance takes place. The term "surface" here also relates to the internal surface of a three-dimensional matrix, for example the internal surfaces of the three-dimensional structure of a zeolite. Examples of adsorbents within the meaning of the present invention are silica gel, activated carbon and aluminosilicates. Aluminosilicates are known to those skilled in the art. The term "aluminosilicates" comprises, for example, acid-activated bentonites (bleaching earths) and zeolites. The bentonites activated with acid (decolorizing earths) are bentonites, the smectites which have been partially dissolved (inflatable or clay minerals) by treatment with acid and which thus have a high surface area and a large micropore volume. The bentonites are clays which have been formed by the wear of volcanic ash (tufa) and consist of the montmorillonite and beidellite minerals (the smectite mineral group). Particularly preferred aluminosilicates in the context of the present invention are zeolites. In this context, those zeolites which do not contain aluminum may also belong to the invention. Zeolites are a widely distributed group of crystalline silicates, more precisely alkali metal containing water or alkaline earth metal aluminosilicates of the general formula M2 / 2? -Al203-xSi02"and H20, wherein M = monovalent or polyvalent metal (usually a cardion of alkali metal or alkaline earth metal) H or NH4, etc., z = the valence of the cation, x = from 1.8 to about 12 ey = from 0 to about 8. The stoichiometric ratio of Si02 to A1203 (modulus) is like an important parameter of zeolites The crystalline network of zeolites is constructed of the tetrahedron of Si0 and A104 which are joined by oxygen bridges, which produces an arrangement in space of equally constructed cavities (adsorption) which are accessible through channels or pore openings, which are equal in size to each other Crystal lattices of this type are able to act as a sieve which admits molecules having a cross section smaller than the pore openings within the cavities of the network, while larger molecules can not penetrate. The zeolites are therefore also called molecular sieves. Electrostatic interactions, hydrogen bonding and other intermolecular forces also play a role in adsorption. Many chemical and physical properties of zeolites are dependent on Al content.

The term zeolites according to the present invention relates not only to natural zeolites but also to synthetic ones. Zeolites of natural origin are formed by hydrothermal conversion from volcanic glasses or deposits containing tufa. According to their crystalline networks, natural zeolites can be classified into fibrous zeolites (eg, mordenite, MOR), foliar zeolites and cubic zeolites (eg, faujasite, FAU and ofendite, OFF). Different zeolites usually give three-letter codes (eg MOR, FAU, OFF). To prepare synthetic zeolites, the starting materials used are Si02-containing substances (for example, soluble glasses, silica-filled, silica sols) and containing Al203 (for example, aluminum hydroxides, aluminates, kaolins) which, together with the alkali metal hydroxides (usually NaOH) they are converted to crystalline zeolites at temperatures above 50 ° C in the aqueous phase. For industrial uses as adsorbents, synthetic zeolites may undergo additional modifications. Preferably, the zeolite should have a pore size of at least 7 Á. Pore size and polarity of zeolites have an influence on distribution weight, for example of different sugars, which give, for example, the separation property in a chromatographic application. The low aluminum zeolites are generally polar and thus of priority for the adsorption of sugars. As already described, zeolites can easily adapt to a separation problem. The primary preparation can affect the pore size, and the polarity can then be varied through a post-treatment by reducing the aluminum content. The preferred zeolites according to the present invention are FAU, BEA and OFF. The properties which are respectively advantageous of different zeolites in the context of the present invention can be seen in Example 1. Particular preference is given to OFF. The enrichment when using aluminosilicate can take place in principle in two different ways. The aluminosilicate can adsorb any undesired foreign matter so that the trehalose remains in solution, or it can adsorb the trehalose so that the unwanted foreign matter remains in solution. In both cases, it is preferable if the adsorption takes place as selectively as possible. As the adsorber, the use can be made of fixed bed, moving bed and fluidized bed adsorbers. The adsorption can be carried out batchwise or continuously. In the mode in which trehalose is adsorbed to aluminosilicate, a number of advantages arise. The number of developmental steps required to isolate trehalose is reduced by selective enrichment of trehalose (in contrast to previous processes to isolate trehalose in which undesired foreign matter with highly varied frequency has to be removed step by step). The number of by-product / waste streams is reduced compared to the removal in stages of unwanted foreign matter. Trehalose, due to selective adsorption, occurs in high purity even after a primary enrichment stage using aluminosilicate. Due to the decreased number of stages of development and the reduced number of by-product / waste streams, production costs are reduced. In addition, trehalose of comparatively low concentration can be cost effectively enriched by selective enrichment. The preferred aluminosilicates, in this embodiment are therefore aluminosilicates, in particular zeolites, to which the trehalose is adsorbed, preferably it binds with high selectivity compared to undesired foreign matter present in the solution.

After trehalose is adsorbed to the aluminosilicate, as an additional step, trehalose can be extracted from aluminosilicate. This is extracted, for example, by eluting with methanol, ethanol, water, hot water (50-100 ° C), hot methanol (50-65 ° C), hot ethanol (50-80 ° C) or other suitable eluents, by example, methylene chloride, acetonitrile, NMP (N-methyl-2-pyrrolidone), DMSO (dimethyl sulfoxide), short chain ketones or short chain ethers. The short chain in this context means a chain length of up to CIO, preferably up to C6, particularly preferably up to C4. A further embodiment of the invention relates to a process for enriching trehalose in which the adsorbent is used in the context of a chromatographic separation. In chromatographic processes, trehalose can be separated through the different activation time behavior compared to other substances present in the solution. This produces fractions with eluates which contain trehalose. Within the meaning of the present invention, the term chromatography comprises all known and suitable chromatographic separation processes, for example, fixed bed chromatography, movable bed chromatography and simulated movable bed chromatography. Chromatography can be carried out batchwise or continuously.

Continuous chromatography can be carried out for example, using Continuous Alternate Ring Chromatography (CRAC), Real Movable Bed Chromatography (TMBC) or Simulated Movable Bed Chromatography (SMB). From the eluate containing trehalose, an enrichment or purification can be performed by means of additional processes which are suitable and known to those skilled in the art. For example, further enrichment or purification of trehalose can take place by precipitation. At this stage, any desired materials of value or unwanted foreign matter may precipitate out. Precipitation can be initiated, inter alia, by adding an additional solvent, adding salt or varying the temperature. The resulting precipitate of solids can be separated by processes known to those skilled in the art. For example, solids can be separated by filtration, such as pressure filtration and vacuum. It is also possible to use cake filtration, deep filtration and cross flow filtration. Preference is given to cross-flow filtration. Particular preference is given here to microfiltration to separate solids > 0.1 μm. An additional possibility for separating solids is sedimentation and / or centrifugation. For centrifugation, various types of constructions can be used, for example, tube and basket centrifuge, especially impeller, by inverting filter centrifuges and disk separators. As an additional enrichment or purification step, the treatment can be carried out with activated carbon or with ion exchangers (anion exchangers and / or cation exchangers). Process steps of this type are known from the prior art (see for example, US 5,441,644, US 5,858,735 and EP 0 555 540 Al). Additional possibilities for enrichment, in particular for purification, are the use of microfiltration and ultrafiltration (for example, cake filtration, depth and cross-flow techniques) and reverse osmosis. In this case, inter alia, microporous, homogeneous, asymmetric and electrically charged membranes can be used, which are produced by known processes. Typical materials for membranes are cellulose, nylon, poly (vinyl chloride), acrylonitrile, polypropylene, polycarbonate and ceramic esters. Membranes may be used, for example, as a plate module, a spiral module, a tube bundle and a hollow fiber module. In addition, the use of liquid membranes is possible. Trehalose can not only be enriched on the feed side and be removed through the retention stream, but also be reduced on the feed side and removed through the permeate / filtrate stream. For further enrichment of trehalose, in particular for purification and final processing, various methods known to those skilled in the art can be used. A preferred process here is crystallization. Crystallization can be achieved, for example, by cooling, evaporation, crystallization under vacuum (adiabatic cooling), crystallization by reaction and salt displacement. Crystallization can, for example, in stirred and non-stirred tanks, in the process of direct contact, in evaporative crystallizers, in vacuum crystallizers in discontinuous form or continuously, for example, in forced circulation crystallizers (Swenson forced circulation crystallizers). or fluidized bed crystallizers (Oslo type). Fractional crystallization is also possible. The crystallization of trehalose is familiar in principle to those skilled in the art and has been extensively described, including crystallization from aqueous solutions (see also columns 4 and 5 in U.S. Patent 5,441,644). For example, crystallization can be achieved, for example by previous ultrafiltration.

A particular typical method for crystallizing trehalose is crystallization by cooling from suitable solvents, for example, ethanol, methanol, water, methylene chloride, acetonitrile, NMP, DMSO, short chain ketones or short chain ethers. The short chain in this context denotes a chain length of up to CIO, preferably up to C6, particularly preferably up to C4. Another crystallization method is precipitation crystallization. In this method trehalose is presented, for example, in water, and then precipitated by adding a solvent of lower solubility, for example, a short chain alcohol or a short chain ketone. The short chain in this context denotes a chain length of up to CIO, preferably up to C6, particularly preferably up to C4. The crystallization can be accelerated by adding small amounts of trehalose crystals, the trehalose crystals act as seeds of crystallization. There are other processes for the additional enrichment of trehalose; in particular, for purification and final processing, there is drying. There are processes for convection drying, for example, drying ovens, tunnel driers, strip desiccators, disk dryers, jet dryers, fluidized bed dryers, air and rotary drum dryers, and spray drying. A preferred process in the context of the present invention is spray drying. Additional processes use contact drying, for example, blade driers. Likewise, the thermal radiation (infrared) and also the dielectric energy (microwave) can be used to dry. An additional field is vacuum or freeze drying. Condensation is also possible, that is, drying which leads to enrichment, but not necessarily to dryness. An additional process for the additional enrichment of trehalose, in particular for the purification and final processing, is nanofiltration. In this process, trehalose is completely or partially retained on the retention side and enriched in this way. It is obvious to those skilled in the art that such enrichment steps can be carried out not only before, but also after the inventive treatment with the aluminosilicate. In a further embodiment, the present invention relates to a process for enriching trehalose from solutions which originate from the enzymatic synthesis of trehalose. Enzymatic synthesis of trehalose is known to those skilled in the art (see, for example, Schiraldi et al. (2002), Trehalose Production: Exploiting Novel Approaches, Trends in Biotechnology, vol.20 (10), pages 421-425 , and also North American Patent 5,919,668 and EP 0 990 704 A2). In an additional embodiment, the solutions are fermentation broths. Fermentation broths within the meaning of the present invention are produced in the culture of eukaryotic and prokaryotic cells. In particular microorganisms (for example, bacteria, yeast or other fungi). The preferred microorganisms in the synthesis of trehalose are Saccharomyces spec. , in particular Saccharomyces cerevisiae; Bacillus spec. , Candida spec, in particular Candida fermentii; Escherichia coli; Corynebacterium spec. , in particular Corynebacterium glutamicum, Corynebacterium acetoacidofirum (for example, ATCC 13870), Corynebacterium lilium (for example ATCC 15990) and Corynebacterium melaseccola (for example ATCC 17965); Pseudomonas spec.; Nocardia spec.; Brevibacterium spec. , in particular Brevibacterium lactofermentum (for example ATCC 13869), Brevibacterium flavum (for example, ATCC 14067), and Brevibacterium divaricatium (for example, ATCC 21642); Arthrobacter spec. , in particular Arthrobacter sulfuris (for example, ATCC 15170), Arthrobacter citoreus (for example, ATCC 11624); Aspergillus spec .; Streptomyces spec .; Microbacterium spec. , in particular Mikrobacterium ammoniaphylum (for example, ATCC 15354); Pichia spec .; Filobasidium spec., In particular Filobasidium floriforme. Additional suitable microorganisms are known to those skilled in the art, see for example, Miyazaki, J.-L., et al. (1996), the accumulation of trehalose by a basidiomycotinous yeast, Filobasidium floriforme. Journal of Fermentation and Bioengineering, vol. 81 (4), pages 315-319. Variants of these strains which are derived by mutation or genetic modification, or which have an increased capacity for synthesis of trehalose, can also be used in the context of the present invention. Microorganisms can also be cultured with the addition of suitable antibiotics, for example, by inducing trehalose synthesis by adding a β-lactam ring antibiotic. In this case, the fermentation broth first comprises not only the cells, but also the culture medium. Depending on the type of fermentation, a significant part of trehalose can accumulate intracellularly. In this case, it is expedient to digest used cells and to extract trehalose using appropriate methods. Suitable methods, for example, ultrasound treatment, detergent treatment, alkaline lysis and / or extraction with alcohol or trichloroacetic acid are known to those skilled in the art (JP 07 000 190, US 5,441,644). In the fermentation broth there are generally considerable amounts of solids which must first be separated preferably. The term solids also comprises cells and cellular constituents such as nucleic acids and proteins in the present context. In order to separate solids, in particular cell constituents, it is advantageous to agglomerate them first. This can be done with any suitable processes, however in this case a solution of the trehalose (eg, by hydrolysis) should be largely avoided. Suitable methods comprise, for example, treatment with alkali, for example, treatment with Ca (OH) 2, or heating. Advantageously, in this case, enzymes having trehalose activity which also occur are possibly also inactivated. The solids can then be separated by processes known to those skilled in the art. Examples of such processes have already been mentioned above. The present process is also suitable for enriching trehalose from solutions, in particular fermentation broths, in which trehalose is present in low concentrations, in particular less than 15 weight percent, measured in the dry weight of the fermentation broth .

Typically, the concentration of trehalose is from 3 to 8% by weight, measured in the dry weight of the fermentation broth. After separating another valuable product, for example, lysine, the mass fraction of trehalose can be increased to 10-20% by weight, measured in the dry weight of the remaining fermentation broth. If the separation of the biomass as an insoluble constituent is also used at the starting point, the concentration of trehalose is then 20-40% by weight, measured in the dry weight of the fermentation broth. Therefore, a further embodiment of the invention is also a process for enriching trehalose from fermentation broths in which trehalose is present at a concentration of less than 15 weight percent, measured in the dry weight of the broth of fermentation. In many fermentations, a plurality of valuable products are produced. Frequently, trehalose is also produced as an additional valuable product. A problem is then that the enrichment or purification processes for substances produced by fermentation are specifically adapted to the respective product of the value (for example, purification through ion exchange chromatography in the case of amino acids or organic acids). After the enrichment of the first valuable product, other valuable products such as trehalose are presently present in an environment which prevents the enrichment of additional value products. An example is the high concentration of ion after extracting amino acids from ion exchange matrices). This is particularly problematic in the case of trehalose, since trehalose does not have special chemical properties (for example, low solubility in aqueous solutions or electrical charge) which are suitable for simple enrichment. Therefore, trehalose is often disposed along with the waste stream from fermentation. It is therefore a further object of the present invention to produce trehalose as an additional valuable product from fermentation broths from which a first valuable product has been or is developed in advance or subsequently. In a further embodiment of the present invention therefore relates to a process for enriching trehalose from an additional product of value from fermentation broths from which at least one first product of value has been or is obtained , which comprises the steps of separating solids and enriching trehalose using an adsorbent, wherein the adsorbent is an aluminosilicate. The present process is distinguished in that it is particularly tolerant towards the properties of the solution in which trehalose is present. Therefore, the inventive process can also be used when trehalose is present in an environment which would usually prevent enrichment. In contrast, the solution in which trehalose is present is treated particularly gently by the present process, so that an additional valuable product can be obtained even after the enrichment of trehalose. Therefore, trehalose can be obtained before, after or at the same time as the first valuable product. Products of value within the meaning of the present invention comprise, for example, organic acids, proteinogenic and non-proteinogenic amino acids, nucleotides and nucleosides, lipids and fatty acids, diols, carbohydrates, aromatics, vitamins and co-factors, storage substances. , for example PHA (polyhydroxyalkanoates) or PHB (polyhydroxybutyrates) and also proteins and peptides (for example, enzymes). A first preferred product of value according to the present invention is the amino acid lysine. In exemplary embodiments, additional processes are shown which are suitable for purifying trehalose from fermentation broths from which another valuable product was obtained in advance. The drawings and examples are for a more detailed illustration of the invention. The attached drawings show, in Figure 1, the selectivity (s) of zeolites for sucrose (sac) and maltose (malt) relative to trehalose (tre). Figure 2 selectivity (s) for sucrose (sac) and maltose (malt) relative to trehalose relative to pore size (p) of selected zeolites. The determination of the pore size; space atom atom center spheres are used representing the van der aals volumes for the atoms, the radius of the spheres corresponding to the radius van der Waals, as defined in the Materials Study of the MSI Program. An expansion factor of 0.9 is applied to the radius van der Waals of the atoms in the pore of the zeolite and the helium atom is then placed in the center of the pore. The expansion factor for helium radium van der Waals is optimized by hand until the expanded space filling volume of the helium atom comes into contact with the spatial fill volumes of the zeolite pore. This helium expansion factor is used as the pore expansion factor (pore size). Figure 3 the selectivity (s) for hydrocarbons in relation to the pore size (p) of selected zeolites ..

Example 1 To compare the diffusion of sugars in several zeolites quantitatively, theoretical calculations are made. In these, conventional dynamic molecular simulations are carried out along a diffusion coordinate. The diffusion coordinate is determined by a small driving force which is applied along the widest pore axis or the widest channel. This simulates the effect of a concentration gradient. A study is made first on whether the stimulation produces qualitatively correct results. For this purpose, the calculated diffusion times for maltose and sucrose in FAU and BEA are compared with the experimental measurements. According to the calculations, maltose diffuses markedly slower than trehalose and sucrose through FAU (see table 1). This is in agreement with the experimental data showing that maltose has a significantly lower adsorption capacity than sucrose. For BEA it is calculated that sucrose, in the context of the period of time used, does not completely migrate through the zeolite (see table 2). This effect (without adsorption) is a general characteristic of other 1-2 disaccharides which were experimentally measured. From these results for BEA and FAU, it is concluded that the calculation produces qualitatively correct predictions for the relative "solubility" of maltose and sucrose in FAU and BEA. First, a list of candidates for suitable zeolites is formed to separate trehalose, maltose and sucrose (table 1).

Table 1: Dynamic molecular simulations are carried out using these zeolites for the 3 sugars. In this way, the relative selectivity of the sugars with respect to diffusion through the corresponding channels can be calculated. Dynamic molecular force field simulations are carried out in a microcanal assembly at 298 K. Relative periods are measured for molecules which are driven through a pore in the structure of the zeolite by electrostatic force. The force is generated by means in which the coordinate of the charged helium atom is fixed on the opposite side of the pore of the molecule, the molecule is then charged uniformly with a corresponding counter-charge on each atom. For example, 5 atoms of trehalose which are narrower to helium is assigned to each, a charge of -0.3 q, while the helium atom has a charge of +1.5 q. The remaining atoms in the system are not charged. The selectivity in Figure 1 is calculated according to the formula below: Selectivity = rea ° '3 where, 8000 s, when tazúoar is > 8000pS The diffusion periods calculated for the sugars are listed in table 2.

Table 2 A graphic representation of the selectivity is shown in Figure 1. From Figure 1, it will be clear that the individual zeolites have different capacities to separate trehalose from a mixture of sugars. OFF (offer) appears to be more versatile, which does not contain aluminum and prefers trehalose markedly compared to the other two sugars. FAU and BEA likewise show a high relative selectivity for trehalose, but also show a certain selectivity for sucrose and maltose.

Example 2 The enrichment of trehalose by precipitation with calcium hydroxide, centrifugation of treatment with subsequent activated charcoal and drying of the residue. 1 1 of lysine fermentation broth is mixed with 250 g of solid calcium hydroxide after the lysine has been separated in an ion exchanger. After the suspension has been stirred for 4 hours, the suspension is centrifuged in a laboratory centrifuge at 3000 g for 10 minutes. As a result of this procedure, 800 ml of a yellowish supernatant is obtained from the intense brown fermentation broth, whose supernatant comprises 7.6 g of the 8 g of trehalose originally used. For additional purification of this supernatant, 400 g of carbon Powdered activated are added. After incubation for 12 hours at room temperature, the activated carbon is separated through a corrugated filter. 650 ml of a slightly yellowish filtrate are obtained, which contains a total of 6.3 g of trehalose. Finally, the filtrate is dried by freezing. The remaining residue of 9.7 g has the trehalose content of 64.9% by weight.

Example 3 Enrichment of trehalose by precipitation with calcium hydroxide, filtration, treatment of subsequent activated charcoal and drying of the residue. In contrast to Example 2, after the precipitation of calcium hydroxide, the solids formed are separated by filtration. This produces 730 ml of a yellowish filtrate. The additional procedure is carried out in a manner similar to Example 2, as a result of which 8.7 g of the dry residue having a trehalose content of 66.2% by weight can be obtained.

Example 4 Enrichment of trehalose by thermally induced precipitation, cross-flow filtration, subsequent activated charcoal treatment and drying of the residue.

Example 5 Enrichment of trehalose by precipitation with calcium hydroxide, centrifugation of subsequent activated charcoal treatment and drying of the residue (broth from the new development). 1 1 of lysine fermentation broth, after the lysine has been separated in an ion exchanger (trehalose content: 11 g / 1), is mixed with 100 g of solid calcium hydroxide. After the suspension has been stirred for 4 hours, the suspension is centrifuged in a laboratory centrifuge at 3000 g for 10 minutes. 20 g of the activated carbon are added to the resulting 800 ml of a dark brown supernatant and the mixture is incubated at room temperature for 19 hours. The activated carbon is separated by filtration. The filtrate contains 8.9 g of trehalose. By concentration in vacuo, 72.6 g of a dark brown sticky residue having a trehalose content of 10.4% by weight are obtained.

Example 6 Enrichment of trehalose by adsorption to activated carbon and desorption with methanol. 100 ml of a fermentation broth containing trehalose (content 9.76 gf / 1) are stirred with 10 g of activated charcoal (CPG 12 x 40) at room temperature for 16 hours. After the mixture is filtered with suction through a slotted sieve filter, the activated carbon is stirred with 100 ml of methanol at room temperature for 60 hours. After renewed filtration, the filtrate is concentrated to dryness in a rotary evaporator. The brown residue of 1.1 g contains 300 mg of trehalose (27% by weight).

Example 7 Enrichment of trehalose by adsorption to activated carbon and removal with ethanol under crystallization by cooling. 300 ml of a solution of trehalose (content 9.25 g / 1) are stirred with 20 g of activated carbon at room temperature for 18 hours. After the mixture is filtered by suction through a slotted sieve suction filter, the activated carbon is mixed with 300 ml of ethanol and stirred under reflux for 15 hours. The activated carbon is filtered while hot and the filtrate is cooled to 0-5 ° C, with the formation of trehalose crystal. After filtering the mixture with suction, 1.3 g of trehalose are obtained as light gray crystals, the filtrate is concentrated to dryness in a rotary evaporator and contains 0.1 g of trehalose as white crystals. The activated carbon, after filtration, is stirred with 300 ml of MeOH at room temperature for 16 hours, filtered and the filtrate is concentrated on a rotary evaporator, as a result of an additional 0.5 g of trehalose obtained as virtually white crystals.

Example 8 Enrichment of trehalose by adsorption to silica gel and removal with methanol. 100 ml of fermentation broth containing trehalose (content 14 g / 1) are stirred with 10 g of silica gel (MR3482) at room temperature for 19 hours.

After the mixture is filtered with suction through a glass suction filter, the silica gel is stirred with 100 ml of methanol at room temperature for 16 hours.

After repeated filtration, the filtrate is concentrated to dryness in a rotary evaporator. The brown residue of 1.5 g contains 110 mg of trehalose (7% by weight).

Claims (11)

1. CLAIMS 1. A process for enriching trehalose from solutions, in which the enrichment is carried out using an adsorbent, wherein the adsorbent is a zeolite.
2. 2. The process as claimed in claim 1, wherein the trehalose is adsorbed to the zeolite.
3. 3. The process as claimed in one of claims 1 to 2, wherein the zeolite is selected from the group consisting of FAU, BEA, DON, EMT, CFI, MOR, MAZ and OFF.
4. 4. The process as claimed in one of claims 1 to 2, wherein the zeolite is chosen from the group consisting of FAU, BEA, EMT, MOR, MAZ and OFF.
5. The process as claimed in one of claims 1 to, wherein the adsorbent is used in the course of a chromatographic process.
6. 6. The process as claimed in one of claims 1 to 5, wherein the solution originates from a synthesis of enzymatic trehalose.
7. The process as claimed in one of claims 1 to 6, wherein the solution is a fermentation broth and the process comprises the step of separating solids.
8. The process as claimed in claim 7, wherein at least one additional product of value apart from trehalose is separated from the fermentation broth.
9. 9. The process as claimed in claim 7 or 8, wherein the fermentation broth originates from a fermentation with at least one microorganism from the group consisting of Saccharomyces spec., Candida spec., Escherichia coli; Corynebacterium spec., Corynebacterium glutamicum, Pseudomonas spec. Nocardia spec .; Brevibacterium spec. , Arthrobacter spec. , Streptomyces spec. , Microbacterium spec., Aspergillus spec., Bacillus spec., Pichia spec. and Filobasidium spec. The process as claimed in one of claims 7 to 9, wherein the trehalose is presented in the fermentation broth at a concentration of less than 15 weight percent measured in the dry weight of the fermentation broth. The process as claimed in one of claims 1 to 10, wherein the process comprises at least one additional step from the group consisting of treatment with activated carbon, ultrafiltration and ion exchange treatment.