EP1984128A1

EP1984128A1 - Metagenome

Info

Publication number: EP1984128A1
Application number: EP07721931A
Authority: EP
Inventors: Christian Uphoff
Original assignee: Georg Fritzmeier GmbH and Co KG
Current assignee: Georg Fritzmeier GmbH and Co KG
Priority date: 2006-02-16
Filing date: 2007-02-16
Publication date: 2008-10-29
Also published as: DE102006007540A1; WO2007093171A1; WO2007093171B1

Abstract

A mixture having enzymatic activity for degrading inorganic and/or organic substances and a method for preparing one of the same are disclosed, where the mixture having enzymatic activity comprises at least one active biocatalyst, in particular an active enzyme and/or an active analogous microorganism which is determined by a metagenomic analysis of a microbiotic mixture, where the microbiotic mixture can be used to degrade inorganic and/or organic substances.

Description

ENZYMATICAL MIXTURE, AND METHOD FOR PRODUCING THIS MIXTURE

The present invention relates to an enzymatic mixture, and a method for producing the same.

It is known in the art that enzymes can be used to promote the biodegradation of organic matter. It is particularly advantageous to add the effective enzymes directly into the systems that are contaminated with organic matter. In particular, the focus has been placed on wastewater treatment plants.

Another type of biodegradation of substances is given by microbiotic mixtures in which with the help of the interaction of specially selected microorganisms degradation of the pollutants is achieved.

However, a disadvantage of this prior art is that, although the microbiotic mixture has better degradation properties, it is expensive to produce. In addition, microorganisms need certain environmental conditions for their survival, which are not always given, which limits their field of application.

Object of the present invention is therefore to overcome these disadvantages.

This object is achieved by an enzymatic mixture according to claim 1, and a method for producing this enzymatically acting mixture according to claim 6.

It is exploited that the effectiveness of a microbiotic mixture is based on the activity of the biocatalysts contained therein, such as the active enzymes. If one determines the effective for the degradation of the substances biocatalysts, especially the active enzymes, one can produce an enzymatic mixture, which has the same active properties, as the microbiotic mixture, but universal can be used. In addition, the biocatalysts and the enzymes can be used even more targeted, since no consideration has to be taken of the interaction of the microorganisms, which bring about a degradation of the pollutants.

It is particularly advantageous if the determination of the active biocatalysts and of the metabolic pathways of the microorganisms present in a microbial mixture known to be effective is carried out by means of membrane analysis.

The metagenome analysis provides a simple, but very efficient method to determine the active biocatalysts, in particular the active enzymes. By means of this metagenome analysis, the genetic information of the microorganisms used is investigated and, due to the naturally given biodiversity, also enzymes and microorganisms that can be used analogously are determined.

The identification of the active enzymes in the microbiotic mixture may be based on the sought enzymatic reaction, i. the measurable activity of an enzyme present in the microorganisms of the microbial mixture or via nucleic acid signatures i. the sequence homology done. Once the biocatalysts have been identified and / or analogously usable enzymes and the microorganisms found, the enzymatic mixture can be assembled. It then exhibits exactly the same mode of action as the microbiotic mixture investigated, but is independent of the interaction of the microorganisms with one another or of the environmental influences on the microbiotic mixture in general.

In this case, in addition to the enzymes and / or the microorganisms, the enzymatically active mixture may also contain other constituents, such as, for example, water.

In addition, a method for producing an enzymatic mixture according to the invention, in which the microbiotic mixture is examined for their active ingredients in a first step. In a subsequent step, at least one biocatalyst, in particular an enzyme and / or a microorganism which has an analogous effect, such as the microbiotic mixture. In a further step, the at least one biocatalyst is converted into an enzymatic mixture.

Particularly advantageous is a method in which the determination of the biocatalyst is carried out by means of metagenome analysis.

Metagenome analysis and the associated metagenome databases, which have evolved in recent years, provide a selection of different biocatalysts. Whether a biocatalyst suitable in its activity is present, which has an analogous mode of action as the investigated microbiotic mixture, can be investigated by means of a so-called activity screening.

In this activity screening, metagenome databases are usually screened with simple disk assays to provide high throughput.

However, it is also possible to perform a sequence-based screening. However, it is not possible to identify completely new structures, since both the sequence-based method and the methods in which PCR or hybridization is used depend on relatively high homology to already known proteins.

In contrast, activity screening usually ensures a high throughput of samples since most of the metagenomic clones show no corresponding activity.

In a particularly preferred embodiment, a mixture of photosynthetic microorganisms and light-emitting microorganisms, the so-called REACRE®, is used as the microbiotic mixture to be investigated. The photosynthetic microorganisms contained in REACRE® are optionally phototrophic, and in particular contain prochlorophytes, cyanobacteria, green sulfur bacteria, purple bacteria, chloroflexus-like forms, heliobacteria or heliobacillus-like forms, or mixtures of two or more thereof. The light-emitting microorganisms contained in REACRE® are Photobacterium phosphoreum, Vibrio fischeri, Vibrio harveyi, Pseudomonas lucifera or Benecka or mixtures of at least two thereof.

This microbial mixture, REACRE®, is particularly interesting because the interaction of light-emitting and photosynthetic microorganisms creates an independence from an external light source. On the other hand, the mixture is almost universally applicable, since their mode of action is very diverse. As an example of the universal substitutability u.a. called: removal of organic and inorganic pollutants in wastewater, degradation of crude oil residues in the soil, treatment of skin diseases, such as eczema, or prevention of algae growth. However, these examples are by no means complete; the enumeration is only intended to show in which different fields the microbiotic mixture can find application.

Since at least partially different enzymes are active in each different application, a targeted use of enzymatically acting mixtures which have as component only the active enzymes are particularly advantageous.

The invention will be further illustrated by the example of the use of REACRE® for the degradation of crude oil residues in the soil.

For this purpose, a bottom section is considered, which is contaminated with crude oil, for example, due to flooding or leaked oil tanks. If REACRE® is added to this floor section, it is possible to observe the degradation of the crude oil. Most of this degradation takes place via the enzymatic effects of the microorganisms themselves.

However, not all enzymes present in the microorganisms are involved in the degradation, but only a certain part. If these active enzymes are determined and converted into an enzymatic mixture, a mixture is obtained which has the same degradation properties as REACRE®, but consists of only a handful of enzymes compared to the entire microbial mixture. To determine these active enzymes, for example, a sample of the bottom section is taken, which is contaminated with crude oil and added to the REACRE®. From this soil sample, the active enzymes or analogously acting enzymes and / or microorganisms are then determined by means of genome analysis. From these enzymes thus determined, an enzymatic mixture can be produced, which has the same mode of action as REACRE®, but is not REACRE®.

Disclosed is an enzymatic mixture for the degradation of inorganic and / or organic substances and a method for producing such, wherein in the enzymatically acting mixture at least one active biocatalyst, in particular an active enzyme and / or an active analogous microorganism, is present is determined via a metagenome analysis of a microbiotic mixture, wherein the microbiotic mixture is useful for the degradation of inorganic and / or organic substances.

Claims

Claims:

1. Enzymatically active mixture for the degradation of organic and / or inorganic substances, characterized in that the enzymatically active mixture comprises at least one biocatalyst for the degradation of the substances, in particular an enzyme and / or a microorganism, the analogous activity properties of a predetermined microbiotic mixture from photosynthetic microorganisms and light-emitting microorganisms, wherein the microbiotic mixture for the degradation of organic and / or inorganic substances is usable.

2. Enzymatic mixture according to claim 1, wherein the at least one biocatalyst present in the enzymatically active mixture, in particular the enzyme and / or the microorganism, is determined by means of metagenome analysis of the microbiotic mixture.

3. Enzymatic mixture according to claim 1 or 2, wherein the microorganisms present in the enzymatically acting mixture are other microorganisms than those in the microbiotic mixture.

4. An enzymatic mixture according to claim 1, 2 or 3, wherein the photosynthetic microorganisms contained in the predetermined microbiotic mixture are optionally phototrophic, and in particular prochlorophytes, cyanobacteria, green sulfur bacteria, purple bacteria, chloroflexus-like forms, heliobacteria or heliobacilliform forms , or mixtures of two or more thereof.

5. Enzymatic mixture according to one of the preceding claims, wherein the light-emitting microorganisms contained in the predetermined microbiotic mixture are photobacterium phosphoreum, vibrio tables, Vibrio harveyi, Pseudomonas lucifera or Benecka or mixtures of at least two thereof.

6. A process for preparing an enzymatic mixture for the degradation of organic or inorganic substances, characterized in that the enzymatically acting mixture has an analogous action of a predetermined microbiotic mixture of photosynthetic microorganisms and light-emitting microorganisms, wherein the microbiotic mixture for the degradation of Substances, the method comprising the steps of:

Examine the microbiotic mixture for its active ingredients;

Determining at least one active biocatalyst, in particular an active enzyme, which has an analogous effect; Producing an enzymatically active mixture comprising the at least one biocatalyst.

7. The method of claim 6, wherein the determination of the at least one effective biocatalyst is carried out by Metagenomanalyse.

8. The method according to any one of claims 6 to 7, wherein the photosynthetic microorganisms contained in the predetermined microbiotic mixture are optionally phototrophic, and in particular prochlorophytes, cyanobacteria, green sulfur bacteria, purple bacteria, chloroflexus-like forms, heliobacteria or Heliobacillus-like forms, or Mixtures of two or more of them.

9. The method according to any one of claims 6 to 8, wherein the light-emitting microorganisms contained in the predetermined microbiotic mixture are Photobacterium phosphoreum, Vibrio fischeri, Vibrio harveyi, Pseudomonas lucifera or Benecka or mixtures of at least two thereof.