KR101598425B1 - Ro1s-5 strain having capability of inhibiting quorum sensing and use thereof - Google Patents

Abstract

The present invention relates to a strain RO1S-5 having quorum sensing inhibitory activity, and more particularly to a strain isolated from a contaminated reverse osmosis membrane and capable of inhibiting quorum sensing, particularly RO1S-5 And a use thereof.
According to the present invention, quorum sensing by bacteria can be suppressed, and particularly, biofilm formation can be effectively suppressed. In addition, the ROIS-5 strain according to the present invention exhibits AHL lactonase activity, and thus can exhibit high substrate characteristics for AHL molecules regardless of the AHL acyl chain length.

Description

(RO1S-5 STRAIN HAVING CAPABILITY OF INHIBITING QUARUM SENSING AND USE THEREOF)

The present invention relates to a strain RO1S-5 having quorum sensing inhibitory activity, and more particularly to a strain isolated from a contaminated reverse osmosis membrane and capable of inhibiting quorum sensing, particularly RO1S-5 And a use thereof.

Quorum sensing (QS) is a technique in which bacteria use signal molecules such as N-acyl-homoserine lactone (AHL) to detect the density of surrounding bacterial populations, (Smith and Iglewski, 2003a). This is the mechanism by which a certain number of bacteria-induced genes are expressed. QS is known to play an important role in many biological processes such as toxic factor production, bioluminescence, antibiotic production, spore formation and biofilm formation (Novick and Geisinger 2008; Ng and Bassler 2009; Williams and Camara 2009).

Among them, biofilm formation is a major cause of clinically important bacterial pathogenesis and is also a major cause of pathogenesis in medical, industrial and environmental processes such as medical implants, oil drilling, paper production, food processing, (2000), which is the most common cause of the decline in the efficiency of the process (Adonizio et al., 2008; Van Houdt and Michiels, 2010; Donlan, 2001; Ammor et al. It is not easy to develop a general technique for controlling biofilm formation, since biofilms can be present everywhere, as well as allowing cells to be resistant to antimicrobials. Therefore, there is a high interest in an effective strategy for controlling such biofilm formation. In particular, QS is an important mechanism for the formation of biofilms.

A QS regulatory mechanism to control biofilm formation, which may interfere with the QS signaling system, such as enzymatic degradation of AHL, inhibit AHL synthesis by S-adenosyl methionine mimetics, or eliminate AHL or compete with AHL receptors And methods of inhibiting QS signaling by AHL-like substances have been studied. The most obvious QS control method may be quorum quenching (QQ) through enzymatic degradation of QS signal molecules by AHL acylase and AHL lactonase. The AHL acylase degrades the AHL molecule in the amide bond of the acyl side chain and produces homoserine and free fatty acids. AHL lactonase is widely conserved among bacteria, degrading AHL molecules by hydrolyzing lactone bonds in homoserine and releasing acyl homoserine. Unlike AHL acylase, which exhibits variable substrate spectra, AHL lactonase is known to be an AHL degrading enzyme that exhibits high substrate properties for AHL molecules regardless of AHL acyl chain length (Dong et al., 2007; Chen et al. al., 2013; Wang et al., 2004). Bacillus sp. After the first detection of the AHL-lactonase coding gene (AIIA) of 240B1, various AHL-lactonases were identified in many bacteria. However, the inhibitory effect of AHL lactonase related to biofilm formation has not been studied yet.

Korean Patent Publication No. 10-2006-0032991

The present invention provides a strain RO1S-5 capable of inhibiting quorum sensing.

The present invention also provides a quorum sensing inhibitor comprising the strain and the culture medium as an active ingredient.

In addition, the present invention provides a method for inhibiting quorum sensing using the strain and a culture thereof.

According to one embodiment of the present invention, RO1S-5 strain having quorum sensing (QS) inhibitory ability deposited with accession number KCTC 18281P can be provided.

The quorum sensing may be a mechanism using AHL (acyl homoserine lactone) as a signal molecule.

The quorum sensing inhibiting ability may be the ability to inhibit biofilm formation.

The ROlS-5 strain may be isolated from the contaminated reverse osmosis membrane.

The RO1S-5 strain may exhibit substrate specificity to AHL (acyl homoserine lactone).

The AHL may be selected from the group consisting of BHL (C4, N-butanoyl-homoserine lactone), HHL (C6, N-hexanoyl-homoserine lactone), OHL (C8, N-octanoyl- , dDHL (C12, N-dodecanoyl-homoserine lactone) and OdDHL (3-oxo-C12, N- (3-oxododecanoyl) -L-homoserine lactone).

The RO1S-5 strain may exhibit AHL (acyl homoserine lactone) lactonase activity.

The RO1S-5 strain may be an AHL (acyl homoserine lactone) degrading enzyme.

The RO1S-5 strain may inhibit biofilm formation by Pseudomonas aeruginosa PAO1.

According to one embodiment of the present invention, a quorum sensing inhibitor comprising the RO1S-5 strain or a culture thereof as an active ingredient can be provided.

The quorum sensing inhibitor may be a biofilm formation inhibitor.

According to one embodiment of the present invention, there is provided a quorum sensing suppression method comprising the step of treating a solid surface with the strain of claim 1 or a culture thereof.

The quorum sensing inhibiting method may be a method for inhibiting biofilm formation.

The solid surface may be a surface made of stainless steel.

The solid surface may be a surface of a vessel or the like.

According to the present invention, quorum sensing by bacteria can be suppressed, and particularly, biofilm formation can be effectively suppressed.

In addition, the ROIS-5 strain according to the present invention exhibits AHL lactonase activity, and thus can exhibit high substrate characteristics for AHL molecules regardless of the AHL acyl chain length.

In addition, the strain RO1S-5 according to the present invention can be used as an application tool capable of controlling the biofilm in an industrial process related to biofouling and the like.

1A to 1D are photographs showing the QQ activity of a strain isolated from a contaminated reverse osmosis membrane by a disk-diffusion assay.
Fig. 2 shows the QQ activity of RO1S-5 analyzed by? -Galactosidase assay.
FIG. 3 is an analysis of the QQ characteristic of RO1S-5 by? -Galactosidase assays. (a) relates to the extracellular activity of RO1S-5, (b) relates to the thermal stability of RO1S-5, and (c) relates to the effect of treatment with Protease K. (d) are the QS-expressing negative control (no AHL) and the positive control (AHL only).
Figure 4a shows a selective ion recording (SIR) profile by LC-MS / MS analysis for OdDHL.
4B shows a selective ion recording (SIR) profile by LC-MS / MS analysis for OdDHL cultured in combination with RO1S-5.
Figure 4c shows a selective ion recording (SIR) profile by LC-MS / MS analysis for 3-oxo-C12-HS.
5A is a structural formula of OdDHL, and 5b is a structural formula of the decomposition product 3-oxo-C12-HS of OdDHL by lactonase.
Figure 6 is an analysis of the RO1S-5 effect on biofilm formation by P. aeruginosa PAO1.

Hereinafter, the present invention will be described in more detail with reference to Examples. The objects, features and advantages of the present invention can be easily understood through the following examples. The present invention is not limited to the embodiments described herein but can be implemented in other forms. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. Therefore, the scope of the present invention should not be limited by the following examples.

Example: Identification of a New Microorganism RO1S-5 Strain

1. Preparation of bacterial strains and media

Among the total 505 bacteria isolated from contaminated reverse osmosis (RO membrane) (Veolia Water Solutions & Technology, Daesan, Chungbuk, Korea), TSB (Difco, Laboratories, Detroit, MI, USA) cultured in the medium, the biofilm inhibitory ability is the selection superior microorganisms were designated as strains RO1S-5 chromotherapy tumefaciens bio LA-raised CV026 (Chromobacterium violaceum CV026;. ATCC 31532, ATCC, Manassas, VA (Luria-Bertani medium, 0.5% yeast extract, 1%) was used as a biosensor strain to test the QQ activity against C4-C8 AHL, and McClean et al. tryptone and 1% NaCl). Agrobacterium rhizogenes NTL4 (pZLR4, Luo et al., 2003) was used as a biosensor for C8-C12 AHL and AB mannitol medium, ATCC medi D-galactopyranoside (Promega, Madison, WI, USA) and gentamycin (gentamycin, Sigma) were added to AB mannitol medium as needed ( Pseudomonas aeruginosa PAO1, ATCC BAA-47) were added to the QS signal (AHLs) producer and the biofilm formers Bacillus cereus ( ATCC 14579) and Escherichia coli DH5α were used as QQ-positive and negative control, respectively (Morohoshi et al., 2009). All three strains were cultured in LB medium at 35 ° C with shaking (150 RPM).

AHLs: BHL (C4, N-butanoyl-homoserine lactone), HHL (C6, N-hexanoyl-homoserine lactone), OHL (C8, N-octanoyl- dDHL (C12, N-dodecanoyl-homoserine lactone) and OdDHL (3-oxo-C12, N- (3-oxododecanoyl) -L-homoserine lactone) were purchased from Sigma-Aldrich. All AHLs stock solutions were prepared using DMSO (Sigma-Aldrich) and stored at -30 ° C. Reagent-grade Proteinase K, ONPG (2-nitrophenyl β-D-galactopyranoside) and other chemicals were purchased from Sigma-Aldrich.

2. Analysis of QS and QQ activity of new microorganisms isolated from contaminated reverse osmosis (RO) membrane

New microorganisms isolated from a contaminated reverse osmosis membrane (RO membrane) collected from a full-scale water treatment plant (Veolia Water Solutions and Technology Korea, Daesan, Chungbuk, Korea, Kim et al., 2009) QS and QQ activities were tested using parallel streaking of cultured cells overnight (Ravn et al, 2001).

The QQ activity of the new microorganism was confirmed by disk-diffusion analysis (Mani et al., 2012). For BHL and HHL detection, CV026 (200 [mu] L) incubated overnight was added to 10 mL fresh LB agar medium at 50 [deg.] C and the agar-culture mixture was immediately poured into a Petri dish. BHL (6.8 μg) or HHL (0.2 μg) mixed with 20 μL of a new microorganism (A600 = 2.6 ± 0.1) cultured in fresh LB medium for 24 hours was prepared and sterilized on a sterilized paper disc (diameter 8 mm; Adventec , Toyo Roshi Kaisha, Ltd. Japan). The plates were incubated for 24 hours and the diameter of colored circles was measured. CV026 was used for the detection of C4 to C8 acyl chains, which exhibit purple color when exposed to exogenous AHLs in shorter acyl chains than C10 except C4-3-oxo AHL (Cha et al., 1998; Farrand et al 2002; Zhu et al., 2003; Kawaguchi et al., 2008).

For detection of dDHL and OdDHL, the same method as BHL and HHL detection was used except that NTL4 cultured for 36 hours as a bioassay strain and AB mannitol agar supplemented with X-Gal and gentamycin were used. The QQ activity of the new microorganism was determined based on the standard curve obtained from the relationship between the amount of AHL and the diameter of the color zone.

It was confirmed by disk diffusion analysis that ROIS-5 in the new microorganism has the highest QQ activity for BHL (C4), HHL (C6), DHL (C10) and OdDHL (3-oxo-C12) Reference). FIG. 1A shows loading of 6848ng of BHL, FIG. 1B shows loading of 199.2ng of HHL, 283.4ng of DHL, and OdHHL (29.7ng) of FIG. 1d. The control (control) is loaded with AHL only. The narrower the color zone, the higher the QQ activity.

In this assay, ROlS-5 did not produce AHL signal molecules that could be detected by CV026 and NTL4. The QQ activity of RO1S-5 was quantitatively analyzed using a? -Galactosidase assay. The activity of? -galactosidase is reported in Tang et al. (2013). NTL4 as a biosensor, DHL (C10), dDHL (C12), and OdDHL (3-oxo-C12) as QH inducing AHL molecules were used. RO1S-5 cultured overnight in TSB agar was inoculated into TSB liquid medium and cultured at 30 ° C. for 12 hours with shaking at 150 rpm. RO1S-5 cultures were divided into three (270 μL) and transferred to three wells (48-well microtiter plates; Falcon, BD, Franklin Lakes, NJ, USA). AHL (either DHL, dDHL or OdDHL) dissolved in 30 μL of DMSO was added to each well and the final concentrations were set to 10 ^-7 , 10 ^-7 , and 10 ^-6 M, respectively. All samples were prepared in triplicate, and 48-well plates were incubated at 30 [deg.] C with shaking for 24 hours. After incubation, the culture was centrifuged at 10,000 xg for 5 minutes and the supernatant was then filter sterilized (pore size of 0.20 mu m; Whatman, Maidstone, Kent, UK). The prepared supernatant was mixed with 1 mM NTL4 culture (1/50 dilution of culture incubated in AB medium for 36 hours) and cultured at 30 占 폚 for 16 hours with shaking. After transferring 200 μL of the culture for 16 hours to the wells of a Honeycomb-plate (Oy Growth Curves Ab Ltd, Piscataway, NJ, USA), 20 μL of ONPG (8 mg / ml in PBS) was added to each well. The honeycomb plates were incubated at 25 ° C for 23 hours using Bioscreen C (Oy Growth Curves Ab Ltd), and the A ₄₂₀ was measured every 30 minutes.

The QQ activity of RO1S-5 was analyzed using a? -Galactosidase assay, and the results are shown in FIG. 2 and FIG. FIG. 2 shows the results of comparing the activities of AHL and RO1S-5 in the presence of AHL alone (relative QS expression = 1.0). Compared with the AHL-only control (100% QS expression), ROIS-5 showed markedly reduced QS-regulated beta -galactosidase expression. This means that the QQ activity of RO1S-5 decreased the available AHLs concentration. The amounts of decrease of β-galactosidase expression by the QQ activity of RO1S-5 against DHL, dDHL and OdDHL were 85%, 97% and 84%, respectively. These results indicate that QQ by RO1S-5 is not affected by substitution of AHL molecules or by acyl chain length.

3. Identification of new microorganism RO1S-5

The 16S rRNA of RO1S-5 was amplified by PCR using a universal primer (27F (5'-AGA GTT TGA TCC TGG CTC AG-3 ') and 1492R (5'-GGT TAC CTT GTT ACG ACT T- Were amplified. Purification (SolGent PCR purification kit, SolGent Co. , Ltd., Korea) and then, PCR product is the ABI PRISM 3730XL DNA analyzer (Applied Biosystems, CA, USA) by sequencing and the sequence of the DNA analyzer ABI Prism ^® 3730xl (Applied Biosystems, CA, USA). This sequence is compared with sequences available in the GenBank database using the BLAST program of the National Center for Biotechnology Information (NCBI) to identify their nearest line and the results are shown in Table 1. RO1S-5 was identified as Bacillus (Bacillus sp.) It shows a sequence and 99% sequence homology of the 16S rRNA parts of Bacillus (Bacillus sp.) CP- H45 ( EU584544) without the bar QQ activity known.

As shown in Table 1, the GenBank accession numbers for RO1S-5 are KJ413087.

Isolate
(Accession No.) Identity of
closest match
(Accession No.) Sequence homology (%) Source of EU584544 RO1S-5
(KJ413087) Bacillus sp. cp-h45 (EU584544) 99 Rolling wastewater from Taiyin
Silk Co. Ltd, Taian, China

Test Example 1: Analysis of the QQ mechanism of RO1S-5

1. Analysis of QQ characteristics of RO1S-5

The extracellular presence of RO1S-5 QQ activity, thermal stability, and the response to proteolytic enzymes were analyzed using the? -Galactosidase assay described above. RO1S-5 cultured overnight in TSB agar was inoculated into TSB liquid medium and cultured at 30 DEG C for 24 hours with shaking at 150 rpm. After the culture, the culture was centrifuged at 10,000 x g for 10 minutes, and then the supernatant was filtered sterilized (pore size of 0.20 mu m; Whatman, Maidstone, Kent, UK) to prepare an extracellular supernatant. Extracellular QQ activity was assayed by performing β-galactosidase assays using the prepared supernatant instead of RO1S-5 cultures (the portion labeled in blue in Figure 11 is the assay that follows). In order to test the thermal stability of RO1S-5 QQ activity, the prepared RO1S-5 supernatant was treated in an autoclave at 121 ° C and 15 psi for 15 minutes and analyzed for QQ characteristics. The prepared supernatant was treated with Proteinase K (final concentration 100 μg / mL) at 37 ° C. for 24 hours and the QQ characteristics of RO1S-5 were analyzed. The results are shown in Fig.

3 (a), in + out indicates intracellular and extracellular activity, and out indicates extracellular activity. As shown in FIG. 3 (a), RO1S-5 showed high extracellular QQ activity against DHL (C10), dDHL (C12) and OdDHL (3-oxo-C12). In FIG. 3 (b), (-) is a sample not subjected to heat treatment and (+) is a sample subjected to heat treatment under the above conditions. The QQ activity of RO1S-5 was found to be weak due to heat treatment, and it was confirmed that heat stability was poor. These results indicate that the QQ activity of RO1S-5 may be an enzyme activity. In FIG. 3 (c), (-) is a sample not treated with Proteinase K and (+) is a sample treated with Proteinase K under the above conditions. It was confirmed that the QQ activity of RO1S-5 is still maintained even when the proteinase K is treated. The results of using 30 μL of DMSO without AHL and 270 μL of RO1S-5 supernatant (no AHL) were negative QS-expression control, and 30 μL of DMSO containing AHL and 270 μL of sterilized TSB medium were used as a positive control.

2. Analysis of QQ mechanism of RO1S-5

To analyze the QQ mechanism, OdDHL was incubated with RO1S-5 and the resulting product was analyzed by UPLC (Waters, Acquity UPLC) and cathodic electrospray ionization (ESI) MS / MS (Waters, Quattro Premier MS). 50 μL OdDHL (dissolved in 1.0 μM 10 ^-2 M in DMSO) was mixed with 450 μL of RO1S-5 12 hr culture. After incubation at 30 ° C for 24 hours, the culture mixture was extracted three times with 500 μL of acidified ethyl acetate and the organic phase was evaporated to dryness under a stream of nitrogen. The residue was reconstituted in 500 μL of a methanol: water (1: 1, v / v) mixture and analyzed by UPLC using a C ₁₈ reversed phase column (Waters, 100 mm x 0.2 mm x 0.17 μm). Methanol (B) containing 0.1% formic acid and 0.1% formic acid was used as the eluent for fractionation at the flow rate of 0.4 ml / min under the following elution conditions: A: B solution at 0 min Continuous ratio change to 100% B solution at 3 minutes starting at ratio 9: 1, maintenance to 4 minutes, continuous ratio change to A: B solution ratio of 9: 1 at 5 minutes, maintained for 6 minutes. The separated samples were analyzed using ESI-MS / MS (Center for zero emission technology, Korea). (3-oxododecanoyl) -L-homoserin) was dissolved in OdDHL (2.5 ㅧ 10 ^-3 M) dissolved in 200 μL DMSO And 300 μL of 1 M NaOH were mixed and allowed to react at 37 ° C. for 6 hours. After the pH of the reaction solution was adjusted to 6.0 with H ₃ PO, the lactone decomposition product was extracted three times with ethyl acetate (Mei et al., 2010). After evaporation of the solvent under a stream of nitrogen for drying, the residue was reconstituted with 500 μL of methanol: water (1: 1 v: v) and used as standard N- (3-oxododecanoyl) -L-homoserine .

AHL lactonase and acylase are known to hydrolyze AHL molecules by hydrolyzing each of the lactone bond in the homoserine lactone ring and the amide bond in the acyl side chain (Dong and Zhang, 2005). Thus, release corresponding to an acyl homoserine molecule or a homoserine lactone containing a free fatty acid as a degradation product indicates whether it is due to the activity of a lactonase or an acylase, respectively. If the degradation product is not detected at all, it means that QQ activity is due to quiescent or antagonist.

The OdDHL standard selective ion recording (SIR) profile by LC-MS / MS analysis is shown in Figure 4a. OdDHL generated a single-molecule ion peak at a mass-to-charge ratio (m / z) of 296 at retention time (RT) of 3.37 min. This corresponds to 97.5% of TIC (total ion current). When OdDHL was incubated with ROIS-5 for 24 hours, only one ion peak was shown in the TIC plot at RT 3.32 min (see top chromatogram in Figure 4c). This was almost the same as the RT result of OdDHL, but there was no ion peak detected at m / z 296 (RT = 3.37 min) (see intermediate chromatogram in FIG. 4C) (See the bottom chromatogram in Figure 4c). These results indicate that OdDHL was completely degraded by RO1S-5 to a compound with m / z 314. This is consistent with the specificity of the acyl homoserine (3-oxo-C12-HS) of OdDHL. To confirm this, 3-oxo-C12-HS was synthesized from OdDHL by hydrolysis of lactone ring and the synthesized ion molecule was compared with that produced by RO1S-5 activity. As shown in Figure 4b, the SIR profile of the synthetic 3-oxo-C12-HS was consistent with the SIR profile of the degradation products of OdDHL by RO1S-5 (see Figure 4c). This result shows that 3-oxo-C12-HS (m / z = 314), in which mass 18 corresponding to a water molecule is increased by the lactone action of RO1S-5 on OdDHL (m / z = 296) (See FIG. 5). 3-oxo-C12-HS was not detected in negative control (OdDHL cultured in TSB medium without RO1S-5). This indicates that the non-enzymatic degradation of the lactone ring, such as alkaline hydrolysis and ethyl acetate effects, during extraction is minimal. In the present invention, the possibility of pH-dependent lactone hydrolysis during culture was excluded by culturing RO1S-5 at a pH of 6.3 to 7.0.

The above results indicate that QQ of RO1S-5 is caused by AHL lactonase activity.

Test Example 2: P. aeruginosa ( P.aeruginosa Analysis of effect of RO1S-5 on biofilm formation by PAO1

Quantification of biofilm formation was performed by a microtiter plate assay (O'Toole and Kolter, 1998). PAO1 cultured overnight in fresh LB medium was diluted to A ₆₀₀ 0.2 and mixed with the supernatant of RO1S-5 (5: 1 V / V) cultured for 24 hours. Then, 1 mL of the mixture was transferred to a polystyrene maker rotor plate 48- (Falcon). After incubation at 30 ° C for 24 hours, the medium was carefully removed and 1 ml of 1% crystal violet solution was added. After 20 minutes, the staining solution was removed and the wells were thoroughly washed with distilled water (0.20 占 퐉 pore size). To quantify the attached cells, crystal violet was dissolved in 95% ethanol and A ₅₉₅ was measured. PAO1 cultures mixed with E. coli were used as a negative QQ control and all samples were prepared in triplicate.

P. aeruginosa is one of the best organisms for screening QQ organisms and compounds because it produces QS signaling molecules (BHL and OdDHL). QS plays an important role in biofilm formation by P. aeruginosa. After confirming the reduction of QS induction by lowering the level of AHL by RO1S-5, the QQ activity of RO1S-5 inhibited the biofilm formation by P. aeruginosa PAO1. Based on the crystal violet microtiter assay And the results are shown in FIG. RO1S-5 effectively inhibited the formation of biofilm by PAO1. PAO1 showed significantly less biofilm in the presence of RO1S-5. And less than about 46.7% when compared to the control without RO1S-5. RO1S-5 has been shown to significantly reduce biofilm formation, which could be used as an application tool to control biofilms associated with biofouling in oncology and industrial processes.

Korea Biotechnology Research Institute KCTC18281P 20140418

Claims (12)

Bacillus sp. (Genus Bacillus) RO1S-5 with quorum sensing inhibition deposited with Accession No. KCTC 18281P.
The method according to claim 1,
The quorum sensing is characterized by using AHL (acyl homoserine lactone) as a signal molecule, and a strain of Bacillus sp. ( Bacillus sp.) RO1S-5.
The method according to claim 1,
The Bacillus sp. Strain Bacillus sp. RO1S-5 is characterized in that the quorum sensing inhibiting ability is a biofilm formation inhibiting ability.
The method according to claim 1,
The RO1S-5 strain is a strain of Bacillus sp. Bacillus sp. RO1S-5, which is characterized by exhibiting substrate specificity against AHL (acyl homoserine lactone).
The method of claim 4,
The AHL may be selected from BHL (N-butanoyl-homoserine lactone), HHL (N-hexanoyl-homoserine lactone), OHL (N-octanoyl-homoserine lactone), DHL (N-decanoyl-homoserine lactone) , dDHL (N- dodecanoyl-homoserine lactone), and Bacillus sp OdDHL characterized in that it comprises at least one selected from the group consisting of (N- (3- oxo-dodecanoyl) -L- homoserine lactone) (Bacillus sp.) RO1S-5 strain.
The method according to claim 1,
The RO1S-5 strain AHL (acyl-homoserine lactone) Lactobacillus Bacillus sp., Characterized in that better represent the activity (Bacillus) RO1S-5 strain.
The method according to claim 1,
The RO1S-5 strain AHL (acyl-homoserine lactone) Bacillus sp. (Bacillus) RO1S-5 strain, characterized in that the indicating decomposition activity.
The method according to claim 1,
The strain RO1S-5 inhibits the formation of biofilm by Pseudomonas aeruginosa PAO1 ( Pseudomonas aeruginosa PAO1), a strain of Bacillus sp. Bacillus sp. RO1S-5.
A quorum sensing inhibitor comprising the strain of claim 1 or a culture thereof as an active ingredient.
The method of claim 9,
Wherein the quorum sensing inhibitor is a biofilm formation inhibitor.
A quorum sensing inhibiting method comprising treating a solid surface with the strain of claim 1 or a culture thereof.
The method of claim 11,
Wherein the quorum sensing suppression method is a biofilm formation suppression method.

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