US 2013/0280731 A1 · RATIONAL ENZYME MINING

Abstract

A method for rational mining for induced enzymes in microbial communities is described. The method is characterized in that a community of microorganisms is provided and that the microbial populations of the community are cultivated in a container under conditions of choice, where the microorganisms are given a defined culturing medium, to eliminate matter deriving from the natural habitat and to allow the microbial community to reach a metabolic steady-state. The method is further characterized in that at least one fraction of microorganisms is taken from the container and transferred into at least two separate containers, where at least one of the fractions of microorganisms is provided with a defined medium which includes an inducing substance and/or a substance against which enzyme/enzymes is/are desired, to induce regulation of expression of the desired enzyme/enzymes, and at least one fraction is provided with a defined medium without said inducing substance and without said substance against which enzyme/enzymes is/are desired, for the purpose of comparison, and the fractions are maintained and/or cultivated. Samples of the two fractions are then withdrawn and analyzed for identification of the induced enzyme/enzymes.

Claims23

Dependent claims

A method for rational mining for induced enzyme/enzymes in microbial communities, comprising the steps of:

(a) providing at least one community of microorganisms,
(f) cultivating the microbial populations of said at least one community in at least one container under conditions of choice, where the microorganisms are given a defined culturing medium to eliminate matter deriving from the natural habitat and to allow the microbial community to reach a metabolic steady-state,
(g) removal from at least one container of step (b) of at least one fraction of microorganisms, and transfer of said at least one fraction into at least two separate containers,
(h) providing at least one of the fractions of microorganisms from step (c) with a defined medium which includes an inducing substance and/or a substance against which enzyme/enzymes is/are desired, to induce a regulation of expression of the desired enzyme/enzymes, and at least one of the other fraction(-s) from step (c) with a defined medium without said inducing substance and without said substance against which enzyme/enzymes is/are desired for the purpose of comparison, and maintaining and/or cultivating the fractions of microorganisms, and
(i) withdrawing and analyzing samples of at least one induced fraction and at least one non-induced fraction of step (d) for identification of the induced enzyme/enzymes.

2 more elements

A method for rational mining for induced enzyme/enzymes in microbial communities and production thereof, comprising the steps of:

(a) providing at least one community of microorganisms,
(b) cultivating the microbial populations of said at least one community in at least one container under conditions of choice, where the microorganisms are given a defined culturing medium to eliminate matter deriving from the natural habitat and to allow the microbial community to reach a metabolic steady-state,
(c) removal from at least one container of step (b) of at least one fraction of microorganisms, and transfer of said at least one fraction into at least two separate containers,
(d) providing at least one of the fractions of microorganisms from step (c) with a defined medium which includes an inducing substance and/or a substance against which enzyme/enzymes is/are desired, to induce a regulation of expression of the desired enzyme/enzymes, and at least one of the other fraction(-s) from step (c) with a defined medium without said inducing substance and without said substance against which enzyme/enzymes is/are desired for the purpose of comparison, and maintaining and/or cultivating the fractions of microorganisms,
(e) withdrawing and analyzing samples of at least one induced fraction and one non-induced fraction of step (d) for identification of the induced enzyme/enzymes,
(f) analyzing the identified enzyme/enzymes, deriving the DNA sequence/sequences that codes for the enzyme/enzymes, and
(h) producing the identified enzyme/enzymes by use of a host organism or the original parental organism.

4 more elements

Dependencies are inferred from the claim text.

Rational enzyme mining

Life of this patent

Rational enzyme mining

Life of this patent

Abstract

Claims23