JP2002020693A - Coating and coated film containing microorganism and process for inhibiting adhesion of organism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a coating which exerts an antifouling effect against large aquatic organisms by producing a biojelly under water by forming a coated film containing microorganisms on an underwater structure. SOLUTION: The coating contains a biojelly-producing microorganism which belongs to genus Alteromonas.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to paints and coatings containing microorganisms that produce bio-jelly, and a method for preventing the attachment of large aquatic organisms using the paints or coatings.

[0002]

2. Description of the Related Art Aquatic organisms such as barnacles, mussels, and algae tend to adhere to ships, fishing nets, and other underwater structures, thereby hindering efficient operation of ships and reducing fuel consumption. This causes problems such as inviting the fishing net, clogging of the fishing net, and shortening the service life. In underwater structures, in addition to large aquatic organisms such as barnacles, mussels, and algae, a slimy thin layer (slime layer) formed by microorganisms and their metabolites in water is also formed. This thin film layer has been removed as one of the contaminants.

[0003] On the other hand, on the other hand, a technique utilizing the fact that large-sized aquatic organisms are unlikely to adhere to an underwater structure on which the above-mentioned thin film layer is formed has been reported. JP-A-8-13
No. 3920 discloses that, among the thin film layers as described above, 0.3
A method for preventing the attachment of large aquatic organisms by forming a bio-jelly layer on the surface of a structure in water is disclosed.

[0004] JP-A-8-81308 and JP-A-8-81308
Japanese Patent Application Laid-Open No. 92009 discloses that the above-mentioned bio-jelly layer can be formed by applying a paint containing a cinnamic acid-based, silane-based, or benzylideneaniline-based compound to an underwater structure.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for forming a coating film containing microorganisms on an underwater structure.
An object of the present invention is to provide a paint and a coating that produce bio-jelly in water and exhibit an antifouling effect on large aquatic organisms, and a method for preventing the attachment of large aquatic organisms using the paint or coating.

[0006]

SUMMARY OF THE INVENTION The present invention provides an Altero
A paint containing a microorganism belonging to the genus monas, the microorganism producing biojelly. The present invention is also a coating film comprising a microorganism belonging to the genus Alteromonas, which contains a microorganism that produces biojelly. The present invention further relates to a method for preventing large aquatic organisms from adhering to a structure in water, wherein the surface of the structure is coated with the coating material or formed with the coating film. It is also an adhesion prevention method. Hereinafter, the present invention will be described in detail.

The present inventors have developed Alterom from biojelly formed using cinnamic acid-based, silane-based, and benzylideneaniline-based compounds as a biojelly-forming agent.
Isolate a microorganism belonging to the genus onas, and found that a paint containing this microorganism exerts an antifouling effect on large aquatic organisms in water when applied to an underwater structure to form a coating film. The present invention has been completed.

[0008] In the present invention, bio-jelly refers to Al
A slime-like or jelly-like substance produced by a microorganism belonging to the genus teromonas, which can prevent large aquatic organisms from adhering when formed on a structure in water. Things.

[0009] Whether or not the above-mentioned bio-jelly is produced can be determined by the following confirmation test. That is,
1 part by weight of a solution containing 3 × 10 ¹⁰ cell / ml microorganisms, 10% by weight of polyacrylamide having a molecular weight of 13,000
70 parts by weight of an aqueous solution, 10 parts by weight of ethylene glycol dimethacrylate and 0.5 part by weight of ammonium persulfate were mixed, and 0.2 parts by weight of TEMED was added. The mixture was applied on an acrylic plate, and allowed to stand at room temperature under a nitrogen atmosphere for 2 hours. To form a film. This is immersed in seawater and left for one month, and a slime-like or jelly-like substance is present on the acrylic plate, and a large aquatic organism is not attached to the bio-jelly-producing microorganism. be able to.

[0010] The large aquatic organism refers to an organism that attaches to a structure in the water and grows to a size that can be visually observed by an individual.
Animals such as mussels, oysters, hydroids, bryozoans, sea squirts, serplas and the like; plants such as Aosa and Shiomidoro can be mentioned.

[0011] The microorganism used in the present invention is not particularly limited as long as it is a microorganism belonging to the genus Alteromonas and produces biojelly, and examples thereof include SHY1-1 strain.
The SHY1-1 strain was isolated by the present inventors from a bio jelly formed on the surface of the filter after immersing a petri dish filled with benzylideneaniline powder, sealed with a filter in seawater at 15 ° C., and two weeks later. And has the property of producing biojelly in an advantageous manner.

The SHY1-1 strain has the form shown in FIG. 1 and is a marine non-fermentative gram-negative bacillus that moves by one polar flagella. In addition, it grows well in a nutrient medium in which sodium chloride has been increased to about the seawater concentration, and produces polysaccharide. The temperature at which growth is possible is 5 to 35C, and the optimum growth temperature is around 30C. Viable pH
Is pH 3 to 9, and the optimum growth pH is around neutrality. As for the culture, shaking culture and stationary culture are possible, but shaking culture is more favorable in growth. In the stationary culture, a thin film that seems to be biojelly is observed. In addition, the SHY1
-1 strain has the bacteriological properties shown in Table 1 below. In the table,
The GC content of the intracellular DNA was measured by the HPLC method. NP indicates that no characteristic colony was formed.

[0013]

[Table 1]

Based on the above properties, "Bergey's Manua
l of Systematic Bacteriology "Krieg, NR and Hol
t, JG, Vol. 1 (1986) Williams & Wilkins and “Bergey's Manual of Determinative Bacteriolog
y "Holt, JG, Krieg, NR, Sneath, PHA, St
aley, JT and Williams, ST, 9th Edition (199
4) From references such as Williams & Wilkins, SHY1-
One strain was identified as a microorganism belonging to the genus Alteromonas. However, the species could not be identified because no characteristic colony pigment was produced. However, microorganisms belonging to the genus Alteromonas, which produce bio-jelly, have not been reported so far. Therefore, the present inventors have designated these microorganisms as new strains and designated them as SHY1-1 strains. Institute of Biotechnology and Industrial Technology, Ministry of Industry and Technology (address: East 1 Tsukuba, Ibaraki Prefecture)
(Chome 1-3), accession number FERM P-17821
The strain was deposited as a strain (deposit date: April 20, 2000).

The microorganism used in the present invention may be FERM P-17821 strain or FERM strain.
A natural or artificial mutant strain of P-17821, other strains belonging to the genus Alteromonas, and the like that can produce biojelly can also be mentioned.

When the above microorganisms are incorporated into a paint and form a coating film on an underwater structure, they produce bio-jelly in seawater and can prevent large aquatic organisms from attaching. As the microorganisms to be mixed in the paint,
The microorganism may be in the form of a culture obtained by culturing the microorganism in a liquid medium or the like, or may be the microorganism itself obtained from the culture by centrifugation or the like.

When incorporated into the above paint, the paint 10
It is preferable to mix 10 ⁴ or more cells in 0 g.
When it is less than 10 ⁴ , when a coating film is formed, large aquatic organisms may attach before the microorganisms grow in seawater to form a bio-jelly layer. Preferably, it is 10 ⁸ or more. The above-mentioned paint is applied to an underwater structure by a roll coater, brush coating, or the like to a thickness of 50 to 200 μm.
It is preferable to apply so that the thickness is 0 μm. More preferably, the thickness is 100 to 500 μm.

The method for immobilizing the microorganisms in the coating film is not particularly limited as long as the microorganisms can survive in the coating film. For example, a carrier binding method in which the microorganisms are bound to an insoluble carrier by a covalent bond or the like; Method: A lattice-type or microcapsule-type inclusive method can be used. As the cross-linking method, a coating film is formed by combining two or more kinds of bio-derived polymers such as polysaccharides (chitin, chitosan, alginic acid, agar, guar gum, etc.) and proteins (milk casein, seratin, polylysine, polyglutamic acid, etc.). Products: ones obtained by crosslinking one or more bio-derived polymers with a crosslinking agent of an aldehyde having two or more functional groups such as glutaraldehyde or a carboxylic acid to form a coating film.

Further, a hydrogel utilizing radical polymerization can be used, and examples thereof include divinylbenzene di (meth) acrylate, bisacrylamide, polyhydroxyethyl methacrylate crosslinked with ethylene glycol dimethacrylate, and polyacrylamide. it can.

In the present invention, the form of the paint containing the microorganism is not particularly limited as long as the paint can survive the microorganism, but the form of the reagent used for immobilizing the microorganism in the coating film is not particularly limited. The selection can be made in consideration of properties and compatibility between the reagent and the microorganism. For example, when polyacrylamide cross-linked by bisacrylamide is used as a film-forming component, the acrylamide monomer itself is harmful to microorganisms, and is not suitable for long-term storage at room temperature or the like in a state where microorganisms and monomers are mixed together. ,
It is preferable that the microorganism and the film-forming component are stored in separate containers and mixed at the time of use to form a coating film of two-pack type or more. In the case where a bio-derived polymer such as a polysaccharide or a protein is used as a coating film-forming component, it may be in the form of a paint in which a bio-derived polymer and a microorganism are contained in the same container.

The paint and coating film of the present invention can be suitably used for underwater structures such as ships and fishing nets, and bio-jelly is produced on underwater structures in water such as the ocean. It is possible to prevent large aquatic organisms such as barnacles, mussels, and blue seaweed from adhering.

[0022]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. Example 1 The SHY1-1 strain (FERM P-17821) was transformed into a liquid medium (10 g / L bactotryptone, 5 g yeast extract /
L, NaCl 24 g / L) with shaking at 30 ° C. for 2 days. The viable cell count in this culture was 3 × 10 ¹⁰ cells / ml according to the viable cell count method. This is called culture solution A. 1 ml of the culture solution A, 70 g of a 10% aqueous solution of polyacrylamide having a molecular weight of 13000, 10 g of ethylene glycol dimethacrylate, and 0.5 g of ammonium persulfate are mixed at 15 ° C. Then, 0.2 g of TEMED is added, and the mixture is coated on an acrylic plate. The film was left at room temperature for 2 hours to form a film. This is referred to as coating film 1.

Example 2 A 5% aqueous ammonia solution 2 was added to 70 g of a 3% aqueous alginic acid solution.
After adding 0 g and stirring, 0.7 ml of the culture solution A and chitosan 3 were added.
50% aqueous solution was added and mixed, applied on an acrylic plate,
It was left at room temperature for 3 hours to form a film. This is called coating film 2.

Example 3 40 g of 2-hydroxyethyl methacrylate, culture solution A
1 ml, 70 g of a 10% solution of NaCl, 5 g of ethylene glycol dimethacrylate, 0.5 g of ammonium persulfate
g was mixed at 15 ° C., then 0.2 g of TEMED was added, and the mixture was applied on an acrylic plate, and left at room temperature for 2 hours in a nitrogen atmosphere to form a film. This is called coating film 3.

Example 4 100 g of a 5% aqueous solution of chitosan, 5 g of NaCl, culture solution A
1.2 ml was added and mixed. Glutaraldehyde (0.5 g) was added to the solution, mixed, applied to an acrylic plate, and left at room temperature for 3 hours to form a film. This is called coating film 4.

Comparative Example 1 A coating film was prepared in the same manner as in Example 1 except that the culture solution A of Example 1 was not added. This is used as a comparative coating film.

Evaluation of antifouling properties The coating films 1 to 4 and the comparative coating film were immersed in a raft (Seto Inland Sea) off the coastal research laboratory of Tamano City Nippon Paint Co., Ltd., and the adhesion of large aquatic organisms was visually observed. Table 2 shows the results. The numbers in the table represent the biofouling area (%). One month later, coating 1
About ~ 4, bio-jelly with a thickness of about 2 mm was observed.

[0028]

[Table 2]

[0029]

According to the present invention, when a paint containing microorganisms is applied to an underwater structure to form a coating film, a bio-jelly is formed on the surface in seawater, and barnacles, mussels, and aosa. Large aquatic organisms can be prevented from adhering to underwater structures.

[Brief description of the drawings]

FIG. 1 is an optical micrograph of SHY1-1 strain.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12N 1/20 C12N 1/20 AZ 11/04 11/04 // (C12N 1/20 (C12N 1 / 20 A C12R 1:01) C12R 1:01) F-term (reference) 4B033 NA12 NB14 NB36 NB48 NB49 NB63 NC05 NC06 NC14 ND07 ND20 4B065 AA01X AC08 BA23 BB12 BD05 CA02 CA60 4H011 AD01 BA01 BB21 BC06 BC19 DA02 DD01 CG DD3 HA366 JA57 KA02 NA05 PB05 PB07

Claims (6)

[Claims] 1. A paint containing a microorganism belonging to the genus Alteromonas, which produces a bio-jelly. 2. The paint according to claim 1, wherein the microorganism is FERM P-17821 strain. 3. A coating film comprising a microorganism belonging to the genus Alteromonas, which comprises a microorganism that produces bio-jelly. 4. The coating film according to claim 3, wherein the microorganism is FERM P-17821 strain. 5. A method for preventing adhesion of large aquatic organisms to a structure in water, wherein the paint according to claim 1 or 2 is applied to a surface of the structure. 6. A method for preventing adhesion of large aquatic organisms to a structure in water, wherein the coating film according to claim 3 is formed on the surface of the structure. .