Probe for detecting bacteria using peptidoglycan-binding protein, and use thereof
The present invention relates to a probe for detecting bacteria using a peptidoglycan-binding protein, and a method for preparing the same. Also, the present invention relates to a method for detecting bacteria using the probe. The probe for detecting bacteria according to the present invention can specifically detect bacteria. That is, the probe according to the present invention can clearly distinguish between yeast and bacteria and can detect both Gram-negative and Gram-positive bacteria, and thus is expected to be usable in various fields as a universal probe for detecting bacteria. Further, use of the probe allows bacteria to be detected by identifying only fluorescence development without an additional enzymatic treatment, thereby enabling a simple and quick bacterial detection. In particular, the probe is expected to be effectively usable in the food industry where quick bacterial detection is required.
1. A probe for detecting bacteria, the probe comprising a peptidoglycan-binding protein (PGBP), a fluorescent material, and a quencher wherein the peptidoglycan-binding protein consists of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 3;
wherein the quencher is linked to the peptidoglycan-binding protein through N-acetylmuramic acid (NAA) or N-acetyl-D-glucosamine (NAG) which is bound to the quencher.
2. The probe according to claim 1 , wherein the fluorescent material is a luminous molecule, a metal ion, an organic dye, a conductor, a semiconductor, an insulator, a quantum dot, or a quantum wire.
3. The probe according to claim 1 , wherein the quencher is any one or more selected from the group consisting of Black Hole Quencher-1 (BHQ-1), 4-(dimethylaminoazo)benzene-4-carboxylic acid (DABCYL), Eclipse, N,N,N′,N′-tetramethyl-6-carboxyrhodamine (TAMRA), QSY-7, Black Hole Quencher-2 (BHQ-2), and Black Hole Quencher-3 (BHQ-3).
4. The probe according to claim 1 , wherein a mixing ratio of the peptidoglycan-binding protein to the quencher is 1:1 to 40 (v/v).
5. A method for preparing a probe for detecting bacteria, the method comprising:
a) binding a fluorescent molecule to a peptidoglycan-binding protein that consists of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 3;
b) preparing a quencher to which N-acetylmuramic acid or N-acetyl-D-glucosamine is bound; and
c) binding the fluorescent molecule-bound peptidoglycan-binding protein in a), with the quencher to which N-acetylmuramic acid or N-acetyl-D-glucosamine is bound in b).
6. A method for detecting bacteria, the method comprising treating a sample with the probe of claim 1 .
7. The method according to claim 6 , wherein the probe binds to a peptidoglycan of bacteria, thereby generating fluorescence.