EP3194559A1

EP3194559A1 - Method and assembly for fermentation

Info

Publication number: EP3194559A1
Application number: EP15813707.5A
Authority: EP
Inventors: Alexander Gross; Michael KÖHLER
Original assignee: Technische Universitaet Ilmenau
Current assignee: Technische Universitaet Ilmenau
Priority date: 2015-11-27
Filing date: 2015-11-27
Publication date: 2017-07-26
Also published as: WO2017088928A1

Abstract

The present invention relates to a method and an assembly for fermentation. In one step of the method an agent providing a substance convertible by fermentation is compartmentalized to form a multiplicity of compartments of a first kind (02). An agent which effects the fermentation is also compartmentalized to form a multiplicity of compartments of a second kind (03). The compartments of the first kind (02) and the compartments of the second kind (03) are arranged in a dispersion medium (01) which preserves the compartments of the first kind (02) and the compartments of the second kind (03) and allows mass transfer at least with the compartments of the first kind (02) or with the compartments of the second kind (03). Conditions to allow fermentation in the dispersion medium (01), in the compartments of the first kind (02) and/or in the compartments of the second kind (03) are produced and the substance convertible by fermentation is therefore converted into a fermentation product.

Description

Method and arrangement for fermentation

The present invention initially relates to a method for

Fermentation in which a material convertible by fermentation is converted to a fermentation product. in the

Furthermore, the invention relates to an arrangement for

Fermentation of a fermentable material.

From the prior art, various solutions for

Co-cultivation of microorganisms of different genera known, for. B. from DE 11 2012 002 557 T5, from the

EP 1 203 811 A2, from US Pat. No. 9,034,632 B2, from the

DE 35 86 346 T2, from DE 102 08 311 AI and from the

DE 44 10 136 AI.

The US 2008/0241877 AI, DE 20 2005 009 425 Ul, the

DE 10 2007 034 580 AI, DE 297 24 255 Ul, the

DE 696 33 189 T2 and DE 10 2014 103 213 Ul show

Process for the cultivation of microorganisms in

closed volumes, some of which can be considered as compartments. The US 2002/0164797 AI, DE 10 2013 018 242 AI, the

DE 10 2013 114 855 AI, WO 03/005023 A2, the

EP 2 204 441 Bl and DE 10 2012 016 951 Al describe methods for fermentation in which starting materials or

Organisms are arranged separately.

Further methods for fermentation are shown in the dissertation by Friedrich, Andrea Sandra: "Studies on Cultivation, Transformation and Fermentation of Wolffia spec", Rheinische Friedrich-Wilhelms-Universität zu Bonn, 2005. In the dissertations of Hauptmann, Ulla: "Continuous co-culture of Tetrahymena thermophila and Pseudomonas putida in a bioreactor" Justus Liebig University Giessen, 2000 and Schmidt, Dorothea Susanne: "Molecular analysis of the probiotic strain Escherichia coli Nissle 1917" University of Dresden, 2009 will present methods for the co-cultivation of various genera in a liquid fermenter. In the scholarly article by Yang, Hengquan et al. :

"Compartmentalization of Incompatible Reagents within

Pickering Emulsion Droplets for One-Pot Cascade Reactions "in Journal of the American Chemical Society 2015, 137, pages 1362-1371, describes a process for the realization of chemical cascade reactions in which incompatible

Starting materials are arranged in compartments;

for example in solids-stabilized drops, oil-in-water emulsions or water-in-oil emulsions.

In the known methods for fermentation prevail approximately uniform conditions in the fermenter. All microorganisms involved experience the same at one time

Ambient conditions and equal concentrations of the

Culture medium contained substances, so that usually only one species for each production step is used. On this basis, it is difficult to realize biotechnological syntheses that contain multi-step synthesis processes, as they occur in nature. Much of the naturally occurring microorganism is existentially dependent on metabolites of other coexisting organisms. The introduction of two or more organisms in a homogeneously operated fermenter is uncontrollable, so that a reproducible product generation is not possible. The object of the present invention, therefore, starting from the prior art is that known from nature

To be able to reproduce processes in a fermentation to a higher degree technically.

Said object is achieved by a method according to the appended claim 1 and by an arrangement according to the appended independent claim 14.

The inventive method is used for technical

Carrying out a fermentation, which in particular

organic substances are converted. It is therefore necessary to provide at least one substance convertible by fermentation, for which purpose a corresponding agent is used according to the invention, which provides this substance, for example, continuously. A method step of the method according to the invention provides for compensating the agent providing the fermentation-exchangeable substance, whereby a plurality of

Compartments of a first kind arises. Both

Compartments are closed

Volume elements that do not coalesce uncontrollably, even when in contact with each other, such that the agent providing the fermentation-alterable agent provides the means

Compartment surface, d. H. the phase boundary is not

can happen.

Furthermore, a fermentation-causing agent

needed to ferment the fermentable material. According to the invention, the fermentation-causing agent is also compacted, resulting in a plurality of compartments of a second type. Both

Compartments of the second kind are also closed volume elements according to the above explanations In a further step of the method according to the invention, the compartments of the first type and the compartments of the second type are arranged in a dispersion medium. The dispersion medium is preferably liquid and is preferably located in an interior of a fermenter. The

Dispersion medium preserves the compartments of the first kind and the compartments of the second kind, so that they do not dissolve and the separate volumes are preserved. Another property of the dispersion medium is that it can transport mass at least with the compartments of either species, i. H. allows mass transport into or out of the compartments of either species, with mass transfer through the dispersion medium or even directly between the compartments. In any case, the dispersion medium in the environment of the compartments offers conditions that the mass transport into or out of the compartments of at least one of the two species is possible. This may, for example, be an indirect or direct mass transport between the compartments of the first type and the compartments of the second type.

But it may also be an indirect or immediate mass transport between the dispersion medium and the

Compartments of the first kind or to act on a direct or indirect mass transfer between the dispersion medium and the compartments of the second kind. Of the

Mass transport can be designed in both directions, so it is a mass transfer. In any case, the mass transport made possible by the dispersion medium is an essential prerequisite for the course of the

Fermentation. The mass transfer includes at least the

Transport of one of the contained components of the compartments of the first kind or at least the transport of one of the

contained components of the compartments of the second kind. The mass transfer is preferably carried out directly or indirectly between the compartments of the first kind and the compartments of the second kind.

According to a further method step are in

Dispersion Medium Conditions for providing fermentation such that the fermentation-alterable material is converted to a fermentation product. The conditions for allowing fermentation are in the dispersion medium

which includes the compartments of the first kind and the compartments of the second kind, since these are in the dispersion medium. Depending on the embodiment, the conditions for granting fermentation are

Dispersion medium and / or in the dispersion medium

to provide located compartments of the first type and / or in the dispersion medium located in the compartments of the second type. Fermentation involves biological processes and / or chemical reactions and / or enzymatic reactions. The fermentation takes place in the dispersion medium, which also contains the compartments of the first type in the dispersion medium and those in the dispersion medium

Includes compartments of the second kind. Thus, the running

Fermentation in the dispersion medium as such and / or in the compartments of the first kind and / or in the compartments of the second kind. Preferably, the fermentation takes place in the compartments of the first kind and / or in the compartments of the second kind.

The generated fermentation product is removed, for example, by removing it from the interior of the fermenter. The removal can be done, for example, continuously or periodically. The discharge of the generated

Fermentation product is preferably formed by recovering the generated fermentation product or by isolating the generated fermentation product. The Fermentation product is preferably from the dispersion medium or from the fermentation product containing

Diverted compartments of the first type or the second type. A particular advantage of the method according to the invention is that it is a biotechnical method

Cultivation under analogous conditions as in spatially separated organized natural systems with a

cooperative metabolic activity in different adapted local milieus.

In preferred embodiments of the invention

Method is the agent providing the fermentable material by a supply of the

Fermentation convertible substance formed. Thus, the material convertible by fermentation is immediately provided and compartmentalized.

In further preferred embodiments of the

The method according to the invention is that the agent providing the fermentation-changeable substance is formed by microorganisms of a first kind which generate the substance convertible by fermentation. Thus, in particular, there is an ongoing production of the fermentable material in the compartments of the first kind

Microorganisms of the first kind may be lysed. The microorganisms of the first kind are preferably formed by living self-regenerating biocatalysts. The fermentable material is preferred

of natural origin and preferably comprises starch and / or sucrose. The fermentable material can also be formed by plant components. The through

But fermentable material can also be transformed by a other dispersible or soluble matter in the agent or in the dispersion medium,

in particular by a natural substance, by an organic compound, by an inorganic compound, by a plant, by an animal, by a fungus, by a mineral or by constituents or combinations of the abovementioned

Examples.

In preferred embodiments of the invention

Method, the fermentation-causing agent is formed by an enzyme or by a catalyst. The enzyme is preferably isolated. However, the agent causing the fermentation can also be replaced by another starting material, such as. B. O ₂ , CO ₂ , H ₂ S, CH ₄ , C ₂ H ₄ or a solution of one of these substances may be formed. The fermentation-causing agent is preferably formed by oxygen.

In further preferred embodiments of the

The method according to the invention is the fermentation

causing agent formed by fermenting microorganisms of a second kind. The microorganisms of the second kind are formed, for example, by bacteria or by yeasts. However, the fermentation-providing agent may also be formed by a lysate of microorganisms.

Preferred embodiments of the method according to the invention comprise a method step in which the

Microorganisms of the first kind and / or the microorganisms of the second kind are first propagated in the compartments, which can be done in particular before the realization of the fermentation. The inventive Kompartimierung allows that the microorganisms of the first kind and / or the

Microorganisms of the second kind in the compartments multiplied can be prevented without undesirable interactions.

In preferred embodiments of the invention

Process, the fermentation is formed by enzymatic reactions and / or biological reactions. Such reactions allow chemoselective conversion of the by the

Fermentation convertible substance and a high concentration of the resulting fermentation product.

The fermentation is preferably formed by a hybrid synthesis, which takes place biocatalytically and chemocatalytically. It is an advantage of the method according to the invention that complex natural metabolic processes in one

Fermentation process in combination with chemically catalytic

Reaction steps can be reproduced technically.

The conditions for granting the fermentation in

Dispersion medium and / or in the dispersion medium

located compartments of the first kind and / or in the im

Compartment medium of the second type of dispersion medium comprise all requirements of a chemical and physical nature which must be present at the end of the fermentation. These conditions are thus on the one hand preferably by physical quantities, such as. Temperature, gas saturation,

Radiation power and pressure or in particular an osmotic pressure formed. On the other hand, the conditions for allowing the fermentation in the dispersion medium and / or in the compartments of the first type and / or in the compartments of the second type are preferably formed by chemical parameters such as pH. The material transport provided according to the invention, at least with the compartments of one of the two types, is preferably mediated by an extractive transport of ingredients via a dispersing carrier phase. This mass transport furthermore preferably takes place by means of a phase transfer process between the compartments of the first type and / or the

Compartments of the second kind.

In preferred embodiments of the invention

The process results in the fermentation before the formation of the fermentation product at least one intermediate product. It is a particular advantage of the method according to the invention that it is the technical realization of several stages

Fermentation processes allows.

The intermediate or the plurality of intermediates

are preferably formed in the compartments of the first kind or in the compartments of the second kind. The intermediate product or the several intermediates are preferably transported into the compartments of the first type or into the compartments of the second type, whereby this transport can take place indirectly via the dispersion medium , Alternatively, the intermediate or the plurality of intermediates are preferably transported into the dispersion medium.

The intermediate or intermediates are preferably formed in the form of compartments of a third kind. Alternatively, the intermediate or intermediates are preferably transported and enriched in compartments of a third type, which is indirectly via the dispersion medium

can be done. The compartments of the third kind thus provide means for collecting the intermediate or the

Intermediates. The several intermediates are preferably formed successively, ie in each case during successive intermediate steps, so that it is a cascade reaction that can be realized with the method according to the invention.

In preferred embodiments of the invention

In the method, the compartments of the first type and / or the compartments of the second type are enveloped with a selective agent which controls the mass transfer between the first and second compartments

Dispersion medium and the compartments of the first type and / or the compartments of the second kind selectively conditional. Thus, a control of the mass transport thereby

be made that the compartments of the first kind and / or the compartments of the second kind with the selective

Envelopes are enveloped to selectively run off the desired fermentation, while undesirable reactions and

Operations are suppressed. The selective agent is

preferably formed by a polymer or by a liquid. The selective agent preferably has a selective one

Permeability to the fermentable material. A criterion of selectivity of the selective

Agent is preferably formed by a molecular weight, by a molecular size, by a polarity, by a lipophilicity or by a charge of the substance to be selected. In the enveloping selective agent is preferably a selective transport such. B. a transport enzyme or a

Detergent, as preferably a phase transfer catalyst or a surfactant introduced. The enveloping of the compartments of the first type and / or of the compartments of the second type with the selective means preferably takes place before arranging the

Compartments of the first kind and the compartments of the second kind in the dispersion medium. In preferred embodiments of the invention

Process produces the fermentation product in the form of

Compartments of a fourth kind, wherein the

Fermentation product can be converted indirectly via the dispersion medium to compartments of the fourth type and enriched, which by separating the

Fermentation product can be made from the dispersion medium.

The fermentation product is formed, for example, by an active ^substance or by an effect substance. The

Fermentation product is preferred by a

Pharmaceutical agent, formed by a dye, by a fragrance or flavor, by a lubricant, by a monomer, by a polymer, by a food or by an antibiotic. Insofar as a polymer than the

Fermentation product is to be obtained, then the

Fermentation process preferred as a multi-stage

Cascade reaction performed in which the individual

Steps proceed alternately in the compartments of the first kind, in the compartments of the second kind or in the compartments of another kind and the dispersion medium, which is preferably controlled by at least one catalyst. Preferably, several stages of the cascade reaction are each controlled by one of the catalysts. The

Fermentation product may also be formed by microorganisms.

The compartments of the first type and the compartments of the second type each have a volume which is preferably less than 1 cm ^{3 in} size; more preferably less than 1 mm ^{3 is} large. The optionally formed compartments of the third type and the optionally formed compartments of the fourth type each have a volume, which is preferably less than 1 cm ^{3 is} large; more preferably less than 1 mm ^{3 is} large.

The compartments of the first type and the compartments of the second type are preferably each formed by a microstructure. The optionally formed compartments of the third kind and the possibly formed compartments of the fourth kind are preferably each formed by a microstructure. The compartments of the first type and the compartments of the second type are preferably each formed microfluidic, d. H. There are microfluidic methods and / or

Microfluidic components used for their preparation. The optionally formed compartments of the third kind, the optionally formed compartments of the fourth kind and the optionally formed compartments of the further type are preferably each formed microfluidic.

The compartments of the first type are preferably each by a particle, by a multiple emulsion, by a

Hollow sphere, formed by a gas bubble, by a capsule or by a drop, in which or in which the fermentation-converting material providing means is arranged.

The compartments of the second type are preferably each by a particle, by a multiple emulsion, by a

Hollow sphere, formed by a gas bubble, by a capsule or by a drop, in which or in which the fermentation-causing agent is arranged.

The compartments of the optionally formed third type are

preferably each by a particle, by a

Multiple emulsion, through a hollow sphere, through a gas bubble, are formed by a capsule or by a drop, in which or in which the intermediate product is arranged.

The compartments of the possibly formed fourth type are

preferably each by a particle, by a

Multiple emulsion, are formed by a hollow sphere, by a gas bubble, by a capsule or by a drop, in which or in which the fermentation product is arranged.

The compartments of the first type and the dispersion medium preferably form an emulsion. Also, the compartments of the second type and the dispersion medium preferably form an emulsion.

The droplets forming the compartments of one or more of the named species are preferably each

solid-stabilized so that they form a so-called Pickering emulsion with the dispersion medium.

The droplets forming the compartments of one or more of the named species are preferably each surfactant-stabilized.

The surface of the droplets forming the compartments of one or more of the named types preferably consists of a medium which is not water-permeable, while the

Dispersion medium consists mainly of water.

The surface of the droplets forming the compartments of one or more of the mentioned types are alternatively

preferably from water, while the dispersion medium

mainly consists of a non-waterable medium. The particles forming the compartments of one or more of the named species are preferably each by a

Gel particles formed by a polymer particle or by an inorganic particle. The particles forming the compartments of one or more of the named species are preferably each formed by a liquid drop, which is enveloped by a further agent, which contains a

ensures selective mass transport from or in the liquid drops. The enveloping further agent is preferably liquid, solid or gel-like.

In preferred embodiments of the invention

Procedure, the individual compartments of at least two of the several species are arranged together in a compartment of a higher order, so that a hierarchical

Topology is formed. For example, the

Compartments of two of the several species can be formed by one drop, wherein the two drops are arranged together in a larger drop, so that a drop-in-drop arrangement is formed.

In a further preferred step of the method according to the invention, the compartments of one or more of the species are microfluidic

Device manipulated to influence the content of the individual compartments.

The dispersion medium allows for direct or indirect transport of a substance which is itself

for example, the fermentation product to one of

Intermediates may be the fermentation-causing agent or the fermentation-alterable agent. The substance may be dissolved or undissolved. Of the

Transportation may be due to chemical reactions and / or based on physical processes. The substance to be transported can, for. B. be formed by oxygen or carbon dioxide. The dispersion medium further preferably allows one

Energy transport from or to the compartments of the first kind and / or the compartments of the second kind

Dispersion medium further preferably allows a

Energy transport between the compartments of the first kind and the compartments of the second kind.

The inventive method preferably further comprises a step in which at least one further the

Fermentation causing agent is compartimated, creating a variety of compartments of a fifth species and possibly other species. In a further step, the compartments of the fifth type and optionally the compartments of the other species are arranged in the dispersion medium. The

Dispersion medium also preserves the compartments of the fifth species and possibly the compartments of the other species. Through this

Embodiments of the method according to the invention can in particular be carried out technically complex fermentations with, for example, in several stages of the fermentation required funds.

The further fermenting agent is preferably formed by an enzyme or by a catalyst. Thus, for example, two different types of

Enzymes are used.

The further fermentation-causing agent is preferably formed by a gaseous substance, such as 0 ₂ , CO ₂ , H ₂ S, CH ₄ , C ₂ H ₄ or a solution of one of these substances. The further fermentation-causing agent is preferred by Formed oxygen. The further fermentation-causing agent is alternatively preferred by a fermentation limiting agent, such as. B. by phosphate ions or by a phosphate ion releasing agent, such as. B. phytic acid formed. The further fermenting agent is preferably constituted by microorganisms of a third kind which are required for culturing the microorganisms of the first kind and / or the microorganisms of the second kind. In this case, co-cultures are preferably formed. The

Microorganisms of the third kind are preferably fermenting.

In a further preferred step of the method according to the invention, the microorganisms of the first kind and the microorganisms of the second kind are combined in the compartments of the first kind, in the compartments of the second kind or in the compartments of the third kind.

In a further preferred step of the method according to the invention, the microorganisms of the first kind and the microorganisms of the third kind in the compartments of the first kind, in the compartments of the second kind, in the compartments of the third kind or in the compartments of the fifth kind united.

In a further preferred step of the method according to the invention, the microorganisms of the second kind and the microorganisms of the third kind in the compartments of the first kind, in the compartments of the second kind, in the compartments of the third kind or in the compartments of the fifth species united.

The described merging of microorganisms of two of the several species in the compartments of one of the species preferably by a change in the composition of

Dispersion medium, controlled by a change in the temperature of the dispersion medium, by a photochemical influence of the dispersion medium or by acting on the dispersion medium electric field. The change of

Composition of the dispersion medium is preferably carried out by adding an emulsion breaker to the respective

To unite microorganisms. The described combination of microorganisms of two of the several species in the compartments of one of the species is preferably controlled by a microfluidic component.

The dispersion medium is preferably formed by water or by a water-immiscible medium in which preferably at least one functional substance is contained. The at least one functional substance preferably comprises at least one detergent. The at least one detergent is preferably by a phase transfer catalyst, by a

Complex images, by an enzyme, in particular by a

Transport enzyme; formed by a surfactant, by a solvent and / or by an emulsion. The dispersion medium is preferably a mineral oil, a silicone oil, a fluorinated solvent, an oil

organic solvent, formed by an aqueous phase, by a polymer solution or by a combination of said substances.

In preferred embodiments of the invention

The method comprises the step of establishing conditions for granting the fermentation in the dispersion medium and / or in the compartments of the first type in the dispersion medium and / or in the dispersion medium in the compartments of the second type at least one substep, in which the conditions are changed as a function of time or as a function of a measured variable describing the course of the fermentation. This change is preferred by a change in the temperature in the

Dispersion medium and / or in the compartments of at least one of the several species or by a change in the

Composition of the dispersion medium formed. The described modification preferably leads to a control of the mass transfer, a control of phase transfer processes and / or of solvent-sensitive transfer processes

and / or a control by phase transfer catalysts and / or by mediators.

The size describing the course of the fermentation is preferably formed by an amount or a concentration of one of the possible starting substances or intermediates or by an amount or a concentration of the fermentation product. The size describing the course of the fermentation is additionally or alternatively preferably formed by a physical quantity or by a chemical quantity. The

physical size is preferably by a temperature of the dispersion medium, by a degree of saturation of a gas, by an osmotic pressure or by an im

Dispersion medium occurring electric field formed. The chemical size is preferably formed by a pH.

In preferred embodiments of the invention

Procedure are sensory compartments in the

Dispersing medium arranged. The sensory compartments convert the size describing the course of the fermentation into an optically measurable signal, so that the sequence of the Fermentation descriptive size from outside the dispersion medium is measurable.

In preferred embodiments of the invention

A method comprises the step of establishing conditions for allowing fermentation in the dispersion medium

at least one sub-step, in which further substances, and / or further of the compartments of the first kind and / or further of the compartments of the second kind, further

Compartments of one of the other species depending on the time and / or depending on the measured the course of the

Fermentation descriptive size are added to the dispersion medium. The procedural step of arranging the compartments of the first type in the dispersion medium is preferably carried out by continuously feeding the compartments of the first kind. The procedural step of arranging the compartments of the second type in the dispersion medium is preferably carried out by continuously feeding the compartments of the second kind.

The continuous feeding of the compartments of the first type into the dispersion medium is preferably carried out depending on the time and / or depending on the measured the course of the

Fermentation descriptive size. This can be certain

Equilibria are ensured in the dispersion medium.

The continuous feeding of the compartments of the second type into the dispersion medium preferably also takes place as a function of time and / or depending on the measured flow of the

Fermentation descriptive size. As a result, certain equilibria can be ensured in the dispersion medium. The procedural step of establishing conditions for granting the fermentation in the dispersion medium preferably comprises at least one partial step in which some of the compartments of the first type and / or some of the compartments of the second type are dependent on the time and / or the course of the Fermentation descriptive size divided into compartment parts, for example, to remove aliquots. The procedural step of discharging the

Fermentation product preferably comprises several substeps. In a partial step, a removal of a portion of the dispersion medium takes place with the compartments therein of the at least two types and the therein

Fermentation product. The removal of this part is preferably carried out depending on the measured the sequence of

Fermentation descriptive size. In another

Partial step is a selection of the fermentation product from the dispersion medium or from the compartments. The selection is preferably carried out depending on the measured

Sequence of fermentation descriptive size. In one

Another sub-step is the dispersion medium,

recycled; especially in the interior of a fermenter. The described selection is preferably not on the

Limited fermentation product. Therefore, in addition to the

Fermentation product preferably also a part of the compartments of the first kind and / or a part of the compartments of the second kind selected. This selection is preferably carried out depending on the measured size describing the course of the fermentation.

The fermentation product is preferably continuous

dissipated and selected. The removal and selection takes place preferably depends on the measured the course of the

Fermentation descriptive size.

In preferred embodiments, it is considered that the compartments of the first type and / or the compartments of the second type may lose their function during the course of the fermentation because, for example, the

corresponding inventories are used up or the

corresponding microorganisms die off. Therefore, be

those of the compartments of the first kind which function as the substance convertible by fermentation

no longer or no longer completely own available means. Likewise those of the

Compartments of the second kind, which no longer or no longer completely possess their function as the fermentation-causing agent. The described selection of compartments of the first type and / or the second type preferably takes place continuously and preferably as a function of the measured variable describing the course of the fermentation.

Particular, preferred embodiments of the method according to the invention qualify it to simulate a natural system. For this it includes further steps. In one of the further steps, there is a compartimization of further agents each providing a substance which can be converted by fermentation, whereby a multiplicity of compartments of other types are formed in each case. The number of further agents each providing a fermentation-alterable agent is equal to the number of other types of compartments and is preferably at least 10; more preferably at least 100. In a further step, there is a Kompartimieren further effecting the fermentation agent, which in each case creates a plurality of compartments of other species. there For example, each of the fermentation-changeable agent-providing agent and the fermentation-causing agent is selected and measured according to a natural system. Thus, the prerequisites are created that the fermentation taking place in the natural system also proceeds in the process according to the invention. Here, the exact sequence of the fermentation, in particular the intermediate steps and the interactions between the one by

Fermentation convertible agent-providing agent and the fermentation-causing agent may not be known. In a further step, arranging also takes place

Compartments of the other species in the dispersion medium, which also preserves the compartments of other species. The described embodiment of the invention

Method that is designed to simulate a natural system can also be extended to study the natural system. For this purpose, some of the means occurring in the natural system are omitted in the compartiming of the further agents each providing a material that can be converted by fermentation. Likewise, when

Compartimizing the further effecting the fermentation

Means some of the resources occurring in the natural system omitted. After the fermentation has been carried out, it is measured which fermentation products have formed.

The process is repeated, then omitting other of the naturally occurring fermentation-providing agent agents and / or other agents causing the fermentation. After the fermentation has been carried out again, it is measured which of the fermentation products were then produced. From the resulting fermentation products can

Conclusions to be drawn, which one by

Fermentation convertible substance-providing agent and / or the fermentation causing agent of

natural system leading to which of the fermentation products.

In preferred embodiments of the invention

Method, the dispersion medium is placed in an interior. The interior is preferred by a

Rührkessel, by a device capable of fermentation, by a capillary gap or by an arrangement of

Micro channels formed.

The arrangement according to the invention is used for fermentation and can therefore generally be referred to as a fermenter. The

Arrangement comprises a means for generating compartments of a first type and a means for generating

Compartments of a second type. The arrangement further comprises an interior space for receiving a

Dispersion medium and at least one feed to the interior for supplying the compartments of the first kind and the

Compartments of the second kind in the interior. In addition, the arrangement comprises means for establishing conditions for allowing fermentation in the interior dispersion medium in the interior space

Compartments of the first kind and / or in the interior compartments of the second kind, so that the convertible by fermentation material is converted into a fermentation product. Another component of the arrangement is formed by a discharge for discharging the fermentation product.

The arrangement according to the invention is preferably designed for carrying out the method according to the invention. The

Arrangement according to the invention is preferred for carrying out one of the preferred embodiments of the invention Process trained. Moreover, the arrangement according to the invention also has such features that are specified in connection with the method according to the invention.

The means for generating compartments are preferred by microfluidic components for the production of droplets, of particles, of multiple emulsions, of hollow spheres, of

Gas bubbles or capsules.

The means for establishing conditions for providing a fermentation preferably comprises a heating and / or a cooling for granting a certain temperature in the interior. The means for establishing conditions for granting a fermentation preferably comprises a device with which a degree of saturation of a gas in the dispersion medium

is adjustable.

In preferred embodiments of the invention

Arrangement is the discharge for discharging the

Fermentation product further formed for discharging the dispersion medium located in the interior. By doing

the dispersion medium to be removed are the

Compartments of the first kind, the compartments of the second kind and the fermentation product. These embodiments further include an associated with the discharge

Separating unit for separating the fermentation product from

Dispersion medium, wherein the separation is preferably carried out depending on a measured the course of the fermentation descriptive size. For detecting the size describing the course of the fermentation, the separating device is preferably equipped with a sensory unit with which this size can be measured. The sensory unit can alternatively or additionally also for the immediate evaluation of the

Dispersion medium and / or the compartments of one of the species be educated. Another component of this

Embodiments is by one with the separation unit

connected return for returning the dispersion medium and / or the compartments formed to the interior.

The separation unit is preferably further designed to be those of the compartments of the first type and / or

those of the compartments of the second kind of

Disperse dispersion medium, which no longer completely have their function as a fermentation-converting agent providing agent or a fermentation-causing agent.

The arrangement according to the invention preferably comprises a metering unit connected to the return, which is designed to meter ingredients into the recirculated compartments.

Further advantages, details and further developments of

Invention will become apparent from the following description of preferred embodiments of the invention, with reference to the drawings. Show it:

Fig. 1: a schematic diagram of an inventive

provided dispersion medium with compartments therein;

2 shows a schematic representation of a cascade reaction in a preferred embodiment of a

inventive method;

Fig. 3: a schematic representation of a stimulation at

preferred embodiments of the method according to the invention; 4 shows a schematic diagram of a product separation in a preferred embodiment of the invention

inventive method;

Fig. 5: a schematic representation of a hybrid synthesis at

a preferred embodiment of the

inventive method; 6 is a schematic representation of a targeted colaence in a preferred embodiment of the invention

inventive method;

Fig. 7: a schematic representation of an enclosure at a

preferred embodiment of the invention

Method;

Fig. 8: a schematic diagram of a use of a

Catalyst in a preferred embodiment of the method according to the invention;

9: a schematic representation of a reaction cascade for

Formation of a polymer in a preferred embodiment of the method according to the invention;

10 shows a schematic representation of the cascade reaction in a further preferred embodiment of a method according to the invention; Fig. 11: a preferred embodiment of a

inventive arrangement for fermentation; and

Fig. 12: a modified embodiment of

Inventive arrangement for fermentation. 1 shows a schematic diagram of a dispersion medium oil according to the invention provided with compartments of a first type 02 and located therein

Compartments of a second species 03. The compartments of the first species 02 contain a culture of a type 1. The

Compartments of the second kind 03 contain a culture of a type 2. The compartments of the first kind 02 and the

Compartments of the second type 03 are in the separation medium

01 dispersed, which is a Stoffaustauch between the

Compartments of the first kind 02 and the compartments of the second kind 03 allows. In the compartments of the first species 02 and in the compartments of the second species 03, microorganisms 06 (shown in Fig. 2) are cultured. The

Compartments of the first type 02 and the compartments of the second type 03 are designed such that the microorganisms 06 can not pass or leave a respective compartment surface 04, which in each case represents a phase boundary. The compartments of the first kind 02 and the

Second type 03 compartments may also be loaded with isolated enzymes, lysed microorganisms, conditioned media or chemical catalysts. The inventive arrangement of the compartments of the first

Art 02 and the compartments of the second type 03 in the liquid dispersion medium 01 allow a parallel co-cultivation of microorganisms in compartments with different starting or media composition for carrying out a fermentation.

Due to the realized mass transport, a substance produced in the compartments of the first type 02 (cf. Fig. 2) are transported into the compartments of the second type 03 and further metabolized there.

Fig. 2 shows a schematic representation of a cascade reaction in a preferred embodiment of the invention

Process. In this schematic diagram that is

Dispersion medium 01 (shown in Fig. 1) not shown. In the compartments of the first type 02 are

Microorganisms of a first species 06. In compartments of a third species 07 are microorganisms of a third kind

08. In compartments of a fourth kind 09 are

Microorganisms of a fourth type 11. By the individual compartmentalization of the involved microorganisms 06, 08, 11 of the fermentation process, comprising stages A, B, C and D, controllable.

Fig. 3 shows a schematic representation of a stimulation in preferred embodiments of the invention

Process. Microorganisms of a fifth kind 13 are

existentially dependent on metabolites of coexisting microorganisms of a sixth species 14 and a seventh species 16 and can be cultivated only in the presence of them or

metabolically stimulate one by fermentation

convertible substance A to a level B. The

Microorganisms of the sixth species 14 and the seventh species 16 may share in compartments of a sixth species 17 or separately in compartments of a seventh species 18 and in

Compartments of an eighth type 19 may be arranged. Alternatively, the microorganisms of the sixth species 14 in their

Compartments of the seventh species 18, the microorganisms of the seventh species 16 in their compartments of the eighth species 19, and microorganisms of an eighth species 21 in compartments of a ninth species 22 in groups in superordinate

Compartments of a tenth type 23 may be arranged. The Microorganisms of the sixth type 14 and the seventh type 16 excrete messengers 24, with which the microorganisms of the fifth type 13 are stimulated. 4 shows a schematic diagram of a product separation in a further preferred embodiment of the invention

inventive method. Ninth-type microorganisms 26 metabolize from stage A to stage B during a fermentation. The metabolized stage B is selectively bound and / or enriched in compartments of an eleventh species 27 and thereby separated.

FIG. 5 shows a schematic representation of a hybrid synthesis in a further preferred embodiment of the invention

inventive method. One tenth of microorganisms

Type 29 metabolizes during a fermentation from a level A over a level B to a level C, where the

Transition from stage B to stage C is controlled by catalysts (not shown) in compartments of a twelfth type 31. The compartimized catalysts allow a hybrid synthesis, which is chemocatalytically from stage A to stage B by enzymes biocatalytically and from stage B to stage C by chemical catalysts. Fig. 6 shows a schematic representation of a targeted

Colaescence in a further preferred embodiment of the method according to the invention. Microorganisms of an eleventh species 33 initially multiply in compartments of a thirteenth species 34. Microorganisms of a twelfth species 36 initially multiply in compartments of a fourteenth species 37. Thereafter, a targeted coalescence of the microorganisms of the eleventh species 33 and the microorganisms of the twelfth species 36 by these are timed to each other and

Compartments of a fifteenth type 38 are generated. Fig. 7 shows a schematic representation of a sheath 39 using the example of the compartments of the first type 02 at a

Another preferred embodiment of the method according to the invention. The envelope 39 consists of a selective

Means passing component A but not component B. The envelope 39 completely surrounds the compartments of the first type 02. Fig. 8 shows a schematic representation of a reaction cascade using a catalyst in a preferred

Embodiment of the method according to the invention. Again, it is a fermentation process, comprising stages A, B, C, starting, by way of example, in the compartments of the first type 02. The reaction step from B to C is carried out using the catalyst. This may be placed, for example, in the fluorinated dispersion medium 01 (shown in Fig. 1) where it selectively effects a reaction step.

Fig. 9 shows a schematic representation of a reaction cascade to form a polymer in a preferred

Embodiment of the method according to the invention. It is a fermentation process comprising stages A, B, C, D, E, and F, exemplified in the compartments of the first kind

02 begins. Step F represents the polymer

Reaction step from B to C and the reaction step from E to F are each carried out using a catalyst. The reaction cascade comprising stages A, B, C, D, E and F alternately runs in the compartments of the first type 02 and in the dispersion medium 01 (shown in Fig. 1). The

Polymerization reaction comes by alternating

Reaction steps that occur sequentially in each case

Compartment 02 and the dispersion medium 01 (shown in Fig. 1) or another compartment run off. In the process, a macromolecule is built up. The exemplary process from stage A to stage F represents only an exemplary part of the repetitive one

Polymerization reaction.

10 shows a schematic representation of the cascade reaction in a further preferred embodiment of a

inventive method. This embodiment is initially similar to the embodiment shown in FIG. Notwithstanding the embodiment shown in Fig. 2 are the

Compartments of the first species 02, the compartments of the third species 07 and the compartments of the four species 09 contacting arranged so that the mass transport in stages B and C directly between the compartments 02, 07, 09 and not indirectly via dispersion medium 01 (shown in Fig 1). The dispersion medium 01 (shown in Fig. 1) is used in this embodiment only for the preservation of

Compartments 02, 07, 09.

Fig. 11 shows a preferred embodiment of a

Inventive arrangement for fermentation. The arrangement comprises a stirred tank 41 forming an internal space. The arrangement furthermore comprises a means (not shown) for producing the compartments of the first type 02 of type A, which can be guided into the stirred tank 41 via a feed 42. In the stirred tank 41 is the dispersion medium 01, in which already the compartments of the second type 03 type C are introduced. The compartments of the second type 03 have catalytic or stimulatory properties, so that there is a fermentation of the substance contained in the compartments of the first type 02, whereby a

Fermentation product of type B is produced, which in

Compartments of a fourth type 43 is included. The Arrangement also includes a device 44, with which the dispersion medium oil with the therein contained compartments of the first, second and fourth type 02, 03, 43 can be mixed. The arrangement further comprises a discharge 46, with which the compartments of the first, second and fourth types 02, 03, 43 and the dispersion medium 01 can be removed from the stirred tank 41. The discharge 46 opens into a sensory unit 47, which serves to measure a size describing the course of the fermentation, and then into a separation unit 48, in which the

Compartments of the fourth type 43 are separated from the compartments of the first and second species 02, 03. In this way, the type B fermentation product can be selected and

be derived. From the separation unit 48 performs a

Return 49 back to the stirred tank 41 so that the

Compartments of the first and second type 02, 03 and the discharged dispersion medium 01 back into the rest

Dispersion medium 01 can be passed in a stirred tank.

The compartments of the first and second types 02, 03 are preferably transported in the feed medium 42, in the discharge 46 and in the return 49 in the dispersion medium 01.

Fig. 12 shows a modified embodiment of

Inventive arrangement for fermentation. In contrast to the embodiment shown in FIG. 7, the embodiment shown here comprises a graduation unit 50 comprising a sensor (not shown) to which is returned

Compartments of the fourth type 43 of type B can be divided, resulting in Teilkompartimente 51 arise. Some of the Teilkompartimente 51 are filled with a metering unit 52 to the volume before selection and fed together with the previously discharged compartments of the first type 02 via the return 49 back to the stirred tank 41. With the embodiment shown here it is possible to selectively select compartments, aliquots of these too

remove and consume unused media components. Thus, analogous to the feed-batch process growth of microorganisms in co-cultures can be realized in a continuous mode of operation.

Another non-illustrated embodiment of the invention provides a co-culture with highly concentrated

Buffers or media before. In the cultivation of

Microorganisms 06, 08, 11 (shown in Fig. 2) and their growth as well as their metabolic conversion changes the composition of the compartments 02, 03 (shown in Fig. 1). The starting composition of compartments 02, 03 (shown in

Fig. 1) must be based on the living conditions of the microorganisms 06, 08, 11 (shown in Fig. 2), which u. a. the maximum concentration of media components, the osmotic pressure and the pH. Thus, growth processes or metabolic production processes are due to the initial one

Composition of the compartments 02, 03 (shown in Fig. 1) limited. The cultivation of a microorganism in

Type A compartments with type B compartments containing media components at a concentration outside of the living conditions of the microorganism allow, with sufficient mass transfer between them

Type A and B compartments improved long term culturing. If the pH changes during the

Cultivation in the compartments of type A becomes the

Type A compartments preferably added a buffer. The

Buffer capacity in the compartments of type A is limited, since too high a salt concentration for the

Microorganism has a toxic or growth-inhibiting effect. The osmotic pressure limits the salt concentration in media. If a compartmented mixed population is used in which the second compartment class contains a buffer system in very high concentration and to a large

Buffer capacity leads, the compartments of type A with the microorganisms can be kept pH stable over a much longer period.

Another non-illustrated embodiment of the invention provides a co-cultivation with different

orthogonal media. The cultivation of different organisms often requires individually adapted

Media conditions. When using z. B.

Conditioned media can thus create the situation that a medium which is responsible for organisms of type A

growth-promoting, but for organisms of type B growth-inhibiting or even toxic. Thus, co-cultivation is not readily possible. By the

However, the use of a compartmentalized approach according to the invention can be two classes of compartments in one

Dispersion medium can be generated. The control of

Substance exchanges by addition of suitable surfactants, phase transfer catalysts or solvents to the separation phase co-culture conditions can be created, which allow a material communication between the compartments, without toxic effects of the first

Dispersion medium on the organisms of type B act.

Another non-illustrated embodiment of the invention provides a screening of multi-organism systems and a direct transfer into the inventive arrangement. It is known from genome analyzes that microorganisms have the genetic makeup, hitherto unknown

To produce secondary metabolites, antibiotics, amino acids or enzymes. The problem is, appropriate Find stimulation conditions to activate these inactive genes. This can be achieved according to the invention by co-cultivation of several organisms. In this case, metabolites or messengers stimulate the formation of the fermentation product. According to a preferred

Embodiment of the invention, for example, 100 different organisms kompartimiert and three each

Compartments Ki, K _j , K _k in each case different

Quantitative ratios n Ki + m _Kj + o K _k co-cultivated. As a result, at least 100 - 99 -98 = 970,200

Possible combinations of different microorganisms, if n, m, o = 1; 1 Ki + 1 K + 1 K _k . It is chosen a suitable combination and the appropriate mixture of

Compartments n Ki + m K _j + o K _k generated in the dispersion medium. Here, the ratio of the compartments n: m: o is set in the same ratio as in the mixture selected in the screening.

Another non-illustrated embodiment of the invention provides a stimulation by diffusion-controlled

Nutrient limitation. Secondary metabolite production is often limited by a limiting factor, such as. B. regulates a phosphate concentration or a nitrogen concentration. This is realized in accordance with a preferred embodiment of the invention by using two compartment classes, one class A containing the production organism and a second class B containing the limiting factor, e.g. B. a source of phosphate or nitrogen in high

Concentration. By the transport control of the limiting factor from the compartments of the class B to the

For Class A compartments, the concentration in the Class A compartments is kept below the required threshold. The limiting factor may also be contained in a depot form in the compartments of type B. For example, be phytic acid as a phosphate source, and transported in this form in the compartments of type A. Another non-illustrated embodiment of

Invention provides an aeration during fermentation. In this case, oxygen or a metabolic gas such as CO _{2 is provided} in physically bound solutions. The oxygen supply of fermentative processes can be realized by injecting sterile air into the fermenter. According to a preferred embodiment of the invention, the oxygen supply of the compartments via the

Controlled oxygen exchange with the dispersion medium.

For example, z. As perfluorinated liquids used, which have a high physical oxygen binding capacity.

Thus, dispersed compartments are supplied with oxygen via the diffusive exchange. The oxygen content in the separation phase is preferably controlled to be a

to realize temporally variable oxygen supply of the compartments. Also preferred are two

Compartment classes used, with a class A the

Production organism contains and a class B, a solution containing the required gas from a chemical and / or

releases enzymatic reaction. These embodiments are not limited to oxygen, but can be combined with any gas, such. As CO ₂ , H ₂ S CH ₄ , C ₂ H ₄ can be realized. In this case, the fermentation process can be designed both aerobically and anaerobically. Another non-illustrated embodiment of

The invention provides an extraction strategy according to which the extractability into the dispersion medium is ensured by an enzymatic transfer of hydrophobic components to a substrate. A problem with biotechnological Production process is the isolation of the product. According to a preferred embodiment of the invention, a bacteriological production step is combined with an enzymatic reaction which alters the product in its polarity so that products are selectively enriched in a compartment class or in the carrier phase. The composition of the dispersion phase is preferably timed to selectively extract products from the compartments.

LIST OF REFERENCE NUMBERS

01 dispersion medium

02 compartments of a first kind

03 compartments of a second kind

04 compartment surface

05

06 microorganisms of a first kind

07 compartments of a third kind

08 microorganisms of a third kind

09 compartments of a fourth species 10

11 microorganisms of a fourth species 12

13 microorganisms of a fifth kind

14 microorganisms of a sixth species 15

16 microorganisms of a seventh species

17 compartments of a sixth kind

18 compartments of a seventh species

19 compartments of an eighth species 20

21 microorganisms of the eighth kind

22 compartments of a ninth kind

23 compartments of a tenth species

24 messengers

25

26 microorganisms of a ninth kind

27 compartments of an eleventh species 28

29 tenth-species microorganisms 30

31 compartments of a twelfth species 32 33 microorganisms of an eleventh species

34 compartments of a thirteenth species

35 -

36 microorganisms of a twelfth species

37 compartments of a fourteenth kind

38 compartments of a fifteenth kind

39 serving

40 -

41 stirred tank

42 feeder

43 compartments of a fourth kind

44 device

45 -

46 exhaustion

47 sensory unit

48 separation unit

49 feedback

50 division unit

51 subcompartments

52 dosing unit

Claims

claims

1. Process for fermentation, comprising the following steps:

- Compartimize a by fermentation

convertible agent providing means (06), whereby a plurality of compartments of a first type (02) is formed;

- Kompartimieren one causing the fermentation

By means of which a multiplicity of compartments of a second kind (03) is produced;

Arranging the compartments of the first kind (02) and the compartments of the second kind (03) in one

Dispersion medium (01), which preserves the compartments of the first kind (02) and the compartments of the second kind (03) and a mass transport at least with the compartments of the first kind (02) or with the

Allows compartments of the second kind (03);

- Create conditions to grant one

Fermentation in the dispersion medium (01), in the

Compartments of the first kind (02) and / or in the compartments of the second kind (03), so that the material convertible by fermentation to a

Fermentation product is converted; and

- Removal of the fermentation product.

2. Method according to claim 1, characterized in that the agent providing the substance convertible by fermentation passes through a reservoir of the product by fermentation

convertible substance is formed. A method according to claim 1, characterized in that the agent providing the substance capable of undergoing fermentation is constituted by microorganisms of a first kind (06) which generate the material convertible by fermentation.

Method according to one of claims 1 to 3, characterized

in that the fermentation-causing agent is formed by an enzyme or by a catalyst.

Method according to one of claims 1 to 3, characterized

in that the fermentation-causing agent is formed by fermenting microorganisms of a second kind.

Method according to one of claims 1 to 5, characterized

characterized in that during the fermentation at least one intermediate product before the emergence of the

Fermentation product is formed.

A method according to claim 6, characterized in that the intermediate product in the compartments of the first kind (02) or in the compartments of the second kind (03) is formed or in the compartments of the first kind (02) or in the compartments of the second kind (03 ) is transported.

A method according to claim 6, characterized in that the intermediate product in the form of compartments of a third kind (07) is formed or in compartments of a third type (07) is transported. Method according to one of claims 1 to 8, characterized

in that the compartments of the first type (02) and / or the compartments of the second type (03) are enveloped with a selective agent, which comprises the

Mass transfer between the dispersion medium (01) and the compartments of the first type (02) and / or the

Compartments of the second kind (03) selectively conditioned.

Method according to one of claims 1 to 9, characterized

in that the compartments of the first type (02) and the compartments of the second type (03) are each formed by a particle, by a hollow sphere, by a gas bubble, by a capsule or by a drop.

Method according to one of claims 1 to 10, characterized in that in the dispersion medium (01)

Functional substances are included, the at least one

Detergent, which by a

Phase transfer catalyst is formed by a surfactant, by a solvent, by a transport enzyme or by an emulsion.

Method according to one of claims 1 to 11, characterized in that the step of producing

Conditions for granting fermentation in

Dispersion medium (01), in the compartments of the first type (02) and / or in the compartments of the second type (03) comprises at least one sub-step, in which the conditions depending on the time or depending on a measured the fermentation descriptive size be changed. Method according to one of claims 1 to 12, characterized in that it is further designed to simulate a natural system, for which it comprises the following further steps:

- Compartimieren further each providing a convertible by fermentation substance providing means, whereby in each case a plurality of compartments of other species is formed;

- Kompartimieren further effecting the fermentation

Means, whereby in each case a multiplicity of compartments of further kinds arises; the others each being a fermentable material

providing funds and the others

Fermentation-causing agents are selected and measured according to a natural system; and

Arranging the compartments of the other species in the

Dispersion medium (oil), which preserves the compartments of the other species.

Arrangement for fermentation, comprising the following components:

- means for generating compartments of a first kind

(02);

- means for generating compartments of a second kind

(03);

- An interior (41) for receiving a

Dispersion medium (01);

- At least one feed (42) to the interior space (41) for supplying the compartments of the first type (02) and the compartments of the second type (03); - A means for establishing conditions for allowing a fermentation in the in the interior (41) located dispersion medium (Ol), in the interior (41) located compartments of the first kind (02) and / or in the interior (41) located Compartments of the second kind (03), so that by fermentation

convertible material is converted to a fermentation product; and

- An outlet (46) for discharging the

Fermentation product.

Arrangement according to claim 14, characterized in that the discharge (46) is furthermore designed to discharge the dispersion medium (01) located in the interior (41), in which the compartments of the first type (02), the compartments of the second type (03 ) and the

Fermentation product are located, the arrangement

further comprises:

a separation unit (48) connected to the discharge for separating the fermentation product from the dispersion medium (01); and

- A return device (49) connected to the separation unit (48) for returning the dispersion medium (01) to the interior space (41).