Enzymatic substrate, synthesis method and uses

An enzymatic substrate of general formula (II):

• in which,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different,
• R5 represents H, NO2, CF3, CN, OCH3, F, I, Cl, Br, SO3H or CO2H in the meta or para position, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzyme.

An enzymatic substrate of general formula (III)

• in which,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different,
• R5 represents H, NO2, CF3, CN, OCH3, F, I, Cl, Br, SO3H or CO2H in the meta or para position, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzyme.

An enzymatic substrate of general formula (IVc):

• in which,
• A represents an aryl or alkyl group,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzyme.

A composition for at least one of detecting, identifying or quantifying at least one microorganism, comprising at least one enzymatic substrate and a reaction medium suitable for said at least one microorganism,

• wherein said at least one enzymatic substrate is of general formula (1):
• in which
• X represents a nitrogen atom or a carbon atom substituted with a phenyl group, said phenyl group optionally being substituted in the meta or para position,
• Y and Z represent a hydrogen atom when X represents a carbon atom, or Y and Z together represent an ether, thioether or amine bond optionally substituted with an alkyl or aryl group,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br and may be identical or different, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzymes wherein R is selected from the group consisting of an α- or β-sugar-type residue, a phosphate and a sulfate.

A method for at least one of detecting, identifying or quantifying at least one microorganism in a specimen, comprising:

• contacting the microorganism originating from the specimen with a reaction medium containing an enzymatic substrate so as to obtain an inoculating medium, and
• detecting the colored or fluorescent product formed by the hydrolysis of said enzymatic substrate,
• wherein said enzymatic substrate is of general formula (I):
• in which
• X represents a nitrogen atom or a carbon atom substituted with a phenyl group, said phenyl group optionally being substituted in the meta or para position,
• Y and Z represent a hydrogen atom when X represents a carbon atom, or Y and Z together represent an ether, thioether or amine bond optionally substituted with an alkyl or aryl group,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br and may be identical or different, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzyme.

Agent for at least one of detecting, identifying or quantifying at least one microorganism, comprising at least one enzymatic substrate and a reaction medium suitable for said at least one microorganism,

• wherein said at least one enzymatic substrate is of general formula (I):
• in which
• X represents a nitrogen atom or a carbon atom substituted with a phenyl group, said phenyl group optionally being substituted in the meta or para position,
• Y and Z represent a hydrogen atom when X represents a carbon atom, or Y and Z together represent an ether, thioether or amine bond optionally substituted with an alkyl or aryl group,
• R1 and R2 each represent, independently of one another, H, Cl, F, I or Br, and may be identical or different, or R1 and R2 together represent a fused benzene ring which may or may not be substituted,
• R3 and R4 each represent, independently of one another, H, Cl, F, I or Br and may be identical or different, and
• R represents a group such that the O—R bond can be hydrolyzed by an enzyme, wherein R is selected from the group consisting of an α- or β-sugar-type residue, a phosphate and a sulfate.