Disinfectant composition, cleaning composition, antifouling composition, virus inactivation composition, and non-bactericidal composition, and composition for removing and suppressing formation of biofilm

Abstract

The purpose of the present invention is to provide a novel disinfectant composition, a novel cleaning composition, a novel antifouling composition and a novel virus-inactivating composition, a novel composition for removing a biofilm and inhibiting the formation of a biofilm, and a novel non-bactericidal composition. According to the present invention, a disinfectant composition, a cleaning composition, an antifouling composition and a virus-inactivating composition each comprising sophorolipid and sodium dodecyl sulfate (SDS) are provided. According to the present invention, a composition for removing a biofilm and / or inhibiting the formation of a biofilm, which comprises sophorolipid and SDS, is also provided. According to the present invention, a non-bactericidal composition containing sophorolipid is further provided.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority from Japanese Patent Application No. 2019-184173 (filing date: October 4, 2019), and the entire disclosure thereof is incorporated herein by reference.

TECHNICAL FIELD

[0002] The present invention relates to a disinfection composition, a cleaning composition, an antifouling composition, and a virus inactivation composition, and a disinfection method, a cleaning method, an antifouling method, and a virus inactivation method. The present invention also relates to a composition for removing biofilm and a composition for suppressing biofilm formation, and a method for removing biofilm and a method for suppressing biofilm formation. The present invention further relates to a non-bactericidal composition.

BACKGROUND ART

[0003] Detergent compositions containing surfactants as active ingredients are widely known, but they are mainly chemically synthesized and their environmental toxicity has become a major problem. On the other hand, although biosurfactants have few safety problems for the human body and the environment, their high cost is a problem in practical applications.

[0004] In addition, the dirt substances include biofilms formed by bacteria. Biofilms are widely found in the environment and cause various problems in people's lives. For example, in the medical field, the formation of biofilms on catheters causes chronic infections, and in the food field, the formation of biofilms on food processing equipment and cooking utensils causes food poisoning, both of which are serious problems in terms of hygiene management. Furthermore, once biofilms are removed, they are often reformed and it is not easy to completely remove them. As detergent compositions for removing or suppressing the formation of biofilms proposed so far, there are those described in Patent Document 1 and Patent Document 2.

[0005] Surfactants are known to be effective in inactivating viruses, and it is thought that surfactants inactivate viruses by destroying the viral envelope (the membranous structure found in virus particles). As virus inactivation compositions containing a surfactant as an active ingredient, those described in Non-Patent Document 2 have been proposed so far.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Non-Patent Documents

[0007]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

[0008] An object of the present invention is to provide a novel disinfection composition and a disinfection method. Another object of the present invention is to provide a novel cleaning composition, an antifouling composition, and a virus inactivation composition, as well as a novel cleaning method, an antifouling method, and a virus inactivation method. A further object of the present invention is to provide a novel composition for removing and suppressing the formation of biofilms, and a novel method for removing and suppressing the formation of biofilms. Yet another object of the present invention is to provide a novel non-bactericidal composition.

[0009] The inventors of the present invention have been intensively researching, paying attention to the surfactant properties of sophorolipids. As a result, they have found that when sophorolipids are used in combination with sodium dodecyl sulfate (which may be referred to as "SDS" in this specification), an excellent biofilm removal effect and an excellent biofilm formation inhibitory effect are achieved. The inventors have also found that when sophorolipids are used in combination with SDS, an excellent virus inactivating effect is achieved. Furthermore, the inventors have found that when sophorolipids are used, a biofilm removal effect and a biofilm formation inhibitory effect are achieved without killing bacteria. The present invention is based on these findings.

[0010] According to the present invention, the following inventions are provided. [1] A disinfecting composition and a disinfectant comprising sophorolipids and sodium dodecyl sulfate (SDS). [2] A cleaning composition and a cleaning agent comprising sophorolipids and sodium dodecyl sulfate (SDS). [3] An antifouling composition and an antifouling agent comprising sophorolipids and sodium dodecyl sulfate (SDS). [4] A virus inactivating composition and a virus inactivating agent comprising sophorolipids and sodium dodecyl sulfate (SDS). [5] The composition and agent according to any one of [1] to [3] above for use in removing and / or inhibiting the formation of biofilms. [6] The composition and agent according to [5] above, wherein the biofilm is derived from Gram-negative bacteria. [7] The composition and agent according to any one of [1] to [6] above, wherein sophorolipids are used at a concentration of 0.1% by mass or less. [8] The composition and agent according to any one of [1] to [7] above, wherein SDS is used at a concentration of 0.1% by mass or less. [9] A composition and an agent for removing and / or inhibiting the formation of biofilms, comprising sophorolipids and sodium dodecyl sulfate (SDS).

[10] A non-bactericidal composition and an agent for removing and / or inhibiting the formation of biofilms, comprising sophorolipids.

[11] The composition and agent according to

[10] above, using sophorolipid at a concentration of 0.1% by mass to 1% by mass.

[12] The composition and agent according to

[10] or

[11] above, for application to an article for which generation of resistant bacteria should be suppressed and / or for application in an environment for which generation of resistant bacteria should be suppressed.

[13] A disinfection method, a cleaning method, an antifouling method, and a method for removing and / or suppressing formation of a biofilm, characterized by using sophorolipid and sodium dodecyl sulfate (SDS).

[14] The method according to

[13] above, wherein the biofilm is derived from Gram-negative bacteria.

[15] A virus inactivation method, characterized by using sophorolipid and sodium dodecyl sulfate (SDS).

[16] The method according to any one of

[13] to

[15] above, using sophorolipid at a concentration of 0.1% by mass or less.

[17] The method according to any one of

[13] to

[16] above, using SDS at a concentration of 0.1% by mass or less.

[18] A method for removing and / or suppressing formation of a biofilm, characterized by using sophorolipid.

[19] The method according to

[18] above, which does not kill bacteria.

[20] The method according to

[18] or

[19] above, using sophorolipid at a concentration of 0.1% by mass to 1% by mass.

[21] The method according to any one of

[18] to

[20] above, applying sophorolipid to an article for which generation of resistant bacteria should be suppressed and / or applying sophorolipid in an environment for which generation of resistant bacteria should be suppressed.

[0011] The disinfectant composition, cleaning composition, antifouling composition, and virus inactivation composition of the present invention can exhibit a disinfection effect, a cleaning effect, and a virus inactivation effect by using a combination of sophorolipid, which is an expensive biological surfactant, and SDS, which is a non-natural surfactant. Therefore, it is advantageous in that it has a low load on the human body and the environment. In addition, the non-bactericidal composition of the present invention can remove biofilms and suppress their formation without killing bacteria by using sophorolipids, which is advantageous in that it reduces the risk of generating resistant bacteria.

Brief Description of the Drawings

[0012]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

[0013] <<Active Ingredients and Uses>> According to the present invention, a disinfection composition comprising sophorolipid and sodium dodecyl sulfate (SDS) as disinfection components is provided.

[0014] According to the present invention, there are also provided a cleaning composition comprising sophorolipid and SDS as cleaning components, an antifouling composition comprising sophorolipid and SDS as antifouling components, and a virus inactivation composition comprising sophorolipid and SDS as virus inactivation components.

[0015] Sophorolipid (sometimes referred to as "SLx" in this specification) is a glycolipid produced during the fermentation of petroleum by yeasts such as Candida species and is classified as one of the "biological surfactants", having low toxicity and excellent biodegradability. Sophorolipid is composed of a hydrophilic region consisting of sophorose, a disaccharide in which two glucoses are linked at the β-1,2 position, and a hydrophobic region consisting of a fatty acid chain in the range of C16 - C18 depending on the type of vegetable oil used during fermentation.

[0016] Sophorolipids include lactone-type and acid-type ones, and a mixture containing either one or both of them can be used in the present invention. Figure 1 shows the chemical structures of the lactone-type and acid-type sophorolipids respectively. The ratio (molar ratio) of the lactone-type sophorolipid to the acid used in the present invention is not particularly limited, for example, it can be 6 - 10:0 - 4, and preferably it can be used at a ratio of about 7:3.

[0017] Sophorolipids produced according to known methods can be used. For example, those produced through the fermentation process of yeast according to JP 2014-150774 A can be used in the present invention.

[0018] SDS is known as a surfactant, and those produced according to known methods or commercially available ones can be used in the present invention.

[0019] In the present invention, "disinfection" means preventing the infection of bacteria and viruses, and is used in the sense of including, in addition to "antibacterial (including bactericidal, sterilizing, and inhibiting the growth of bacteria)" and "virus inactivation", preventing bacterial infection through biofilm removal and inhibition of biofilm formation.

[0020] In the present invention, "washing" is used in the sense of including, in addition to washing to remove dirt, sterilizing (including bactericidal) to reduce bacteria, deodorizing (including odor elimination) to reduce odors, and bleaching to remove pigments. That is, the washing composition is used in the sense of including a detergent, a sterilizing agent (including a bactericidal agent), a deodorizing agent (including an odor eliminating agent), and a bleaching agent. In the present invention, "anti-fouling" means suppressing or preventing the adhesion of dirt. In the present invention, "dirt" also includes foreign substances including organisms and microorganisms.

[0021] Since sophorolipids and SDS have the effects of biofilm removal and biofilm formation inhibition as described in the following examples, the cleaning composition of the present invention containing sophorolipids and SDS can be used in any or both of the forms where biofilm removal and biofilm formation inhibition are expected, and the antifouling composition of the present invention can be used in the form where biofilm formation inhibition is expected.

[0022] Here, a biofilm is composed of extracellular polymeric substances secreted by bacteria and is also called a biological membrane or slime. The type of biofilm targeted in the present invention is not particularly limited, and examples include those derived from Gram-negative bacteria and those derived from Gram-positive bacteria. Examples of Gram-negative bacteria include Pseudomonas aeruginosa ( Pseudomonas aeruginosa ), Pseudomonas putida ( Pseudomonas putida ), etc., bacteria belonging to the genus Pseudomonas ( Pseudomonas ), Escherichia coli ( Escherichia coli ), Alcaligenes ( Alcaligenes faecalis ), Klebsiella ( Klebsiellapneumoniae ), Proteus ( Proteus vulgaris ), Serratia ( Serratia marcescense ), Methylobacterium ( Methylobacterium ), bacteria belonging to the genus Saccharomyces ( Saccharomyces ), bacteria belonging to the genus Rhodotorula ( Rhodotorula ), fungi belonging to the genus Pichia ( Pichia ), bacteria belonging to the genus Sphingomonas ( Sphingomonas ), bacteria belonging to the genus Klebsiella ( Klebsiella ), bacteria belonging to the genus Flavobacterium ( Flavobacterium ), bacteria belonging to the genus Roseomonas ( Roseomonas ). Examples of Gram-positive bacteria include Staphylococcus epidermidis ( Staphylococcus epidermidis ), Acne bacilli ( Propionibacterium acnes ), Staphylococcus aureus ( Staphylococcus aureus) can be mentioned. The biofilm targeted in the present invention may also be derived from resident bacteria separated from the skin (including the scalp).

[0023] Furthermore, the target substrate on which a biofilm may be formed is not particularly limited, and examples are as follows. · Medical instruments (e.g., surgical instruments such as scalpels, forceps, scissors, tweezers, stents, catheters, endoscopes, hoses of dental treatment units) · Water treatment facilities (e.g., reverse osmosis membranes, water supply pipes, drain pipes, water storage tanks) · Air conditioning facilities (e.g., air conditioning equipment such as air conditioners, drain pans, drain pipes, filters) · Ship hulls and port facilities (e.g., ship bottoms, anchors, screws, shafts, piers) · Fishing gear (e.g., fishing nets, stationary nets, aquaculture nets) · Hot spring and bath facilities (e.g., pools, bathhouses) · Kitchen utensils (e.g., cooking utensils such as knives, cutting boards; tableware such as spoons, chopsticks, plates, cups, bowls, bowls) · Food processing equipment (e.g., cutters, slicers) · Foods (e.g., vegetables, fruits) · Food packaging containers (e.g., trays for livestock meat and fresh fish) · Parts of the living body (e.g., skin, scalp, hair, oral cavity) · Daily necessities (e.g., glasses, dentures, false teeth) · Textile products (e.g., clothing) · Residential facilities (e.g., floors, carpets, walls, bathrooms, toilets, washrooms, faucets, handrails, doorknobs) · Furniture (e.g., desks, tables, chairs, sofas) · Daily necessities

[0024] Since sophorolipids and SDS have the effect of virus inactivation as described in the examples below, the virus inactivation composition of the present invention can be used in a form in which virus infection prevention is expected, that is, in a form of disinfecting the virus. Here, in the present invention, "virus inactivation" means losing or weakening the infectivity of the virus.

[0025] The target substrate on which a virus may be present or attached is not particularly limited and is common to the target substrate on which a biofilm may be formed.

[0026] The type of virus targeted in the present invention is not particularly limited, and examples include enveloped viruses. Examples of enveloped viruses include viruses belonging to the Coronaviridae family (e.g., SARS-CoV-2, SARS-CoV-1, MERS), viruses belonging to the Orthomyxoviridae family (e.g., influenza A virus, influenza B virus), and viruses belonging to the Paramyxoviridae family (e.g., measles virus, RSV).

[0027] <<Disinfectant composition>> The disinfectant composition of the present invention can be provided as a composition for disinfecting a target substrate including the above target substrate, a target substrate where the presence and formation of a biofilm are not desired, and a target substrate where a virus is present and attached. Specifically, when the disinfectant composition of the present invention is for the purpose of disinfecting medical instruments, it can be provided as a medical instrument disinfectant; when it is for the purpose of disinfecting kitchen utensils, it can be provided as a kitchen disinfectant; when it is for the purpose of disinfecting the skin, it can be provided as soap, hand soap, or body soap; when it is for the purpose of disinfecting the oral cavity, it can be provided as an oral disinfectant (e.g., mouthwash); when it is for the purpose of disinfecting dentures and removable teeth, it can be provided as a denture disinfectant and a removable tooth disinfectant; when it is for the purpose of disinfecting a bathroom, bathtub, and bathroom supplies, it can be provided as a bathroom disinfectant; when it is for the purpose of disinfecting a toilet, it can be provided as a toilet disinfectant. Since various problems occur in the target substrate due to the formation of a biofilm and the attachment of a virus, the disinfectant composition in the above aspect is expected to remove and suppress the formation of a biofilm from the target substrate and inactivate the virus. Further, when applied to a living body, it can be applied to mammals including humans, for example. When the disinfectant composition of the present invention is also intended for cleaning, antifouling, etc. in addition to disinfecting the target substrate, it can also be used as a disinfectant cleaner or a disinfectant antifouling agent.

[0028] The disinfection composition of the present invention exhibits a disinfection effect with two components, sophorolipid and SDS, but may further contain components generally formulated in the disinfection composition according to the object to be disinfected, and the following cleaning components and the following other optional components can be formulated. For example, when the disinfection composition of the present invention is provided in the form of a medical instrument disinfectant, in addition to sophorolipid and SDS, it can be provided as an alkaline disinfectant formulated with an alkaline agent such as potassium hydroxide or sodium hydroxide, or, in addition to sophorolipid and SDS, it can be provided as an enzyme disinfectant formulated with a proteolytic enzyme and a surfactant. Also, when the disinfection composition of the present invention is provided in the form of a bactericide or a bleaching agent, in addition to sophorolipid and SDS, a bactericide or a bleaching agent (for example, a chlorine-based bleaching agent, an enzyme-based bleaching agent) can be formulated.

[0029] The disinfection composition of the present invention can be used so as to achieve a specific sophorolipid concentration in its usage mode. The lower limit value of the sophorolipid concentration in this case can be, for example, 0.00001% by mass, 0.0001% by mass, or 0.001% by mass, and the upper limit value can be, for example, 1% by mass, 0.1% by mass, or 0.01% by mass. These lower limit values and upper limit values can be arbitrarily combined respectively, and the range of the sophorolipid concentration described above can be, for example, 0.00001 to 1% by mass or 0.0001 to 0.1% by mass. The disinfection composition of the present invention can also be used so as to achieve a specific SDS concentration in its usage mode. The lower limit value of the SDS concentration in this case can be, for example, 0.0001% by mass, 0.001% by mass, or 0.01% by mass, and the upper limit value can be, for example, 1% by mass or 0.1% by mass. These lower limit values and upper limit values can be arbitrarily combined respectively, and the range of the SDS concentration described above can be, for example, 0.0001 to 1% by mass or 0.001 to 0.1% by mass.

[0030] The contents of sophorolipid and SDS in the disinfection composition of the present invention can be arbitrarily set according to the usage form of the disinfection composition by referring to the above usage concentrations. That is, the contents of sophorolipid and SDS in the disinfection composition of the present invention can be determined so that they are at effective concentrations in the assumed usage form. When the disinfection composition of the present invention is provided in the form of a liquid medical instrument disinfectant (assuming 200-fold dilution during use), for example, the concentration of sophorolipid can be 0.002 to 20% by mass, and the concentration of SDS can be 0.02 to 20% by mass.

[0031] <<Cleaning composition>> The cleaning composition of the present invention can be provided as a composition for cleaning a target substrate in which the presence and formation of biofilm are not desired, including the above target substrate. Specifically, when the cleaning composition of the present invention is for cleaning medical instruments, it can be provided as a medical instrument cleaner; when it is for cleaning a ship's hull, it can be provided as a ship's hull cleaner; when it is for cleaning kitchen utensils, it can be provided as a kitchen detergent; when it is for cleaning the skin, it can be provided as soap, hand soap, or body soap; when it is for cleaning the scalp and hair, it can be provided as shampoo; when it is for cleaning the oral cavity, it can be provided as a dentifrice and an oral cleaner (for example, mouthwash); when it is for cleaning glasses, it can be provided as a glass cleaner; when it is for cleaning dentures and removable teeth, it can be provided as a denture cleaner and a removable tooth cleaner; when it is for cleaning textile products such as clothing, it can be provided as a laundry detergent; when it is for cleaning a bathroom, a bathtub, and bathroom supplies, it can be provided as a bathroom detergent; when it is for cleaning a toilet, it can be provided as a toilet detergent. Since various problems occur due to the formation of biofilm, removal and formation inhibition of biofilm from the target substrate are expected in the cleaning composition of the above aspect. Further, when applied to a living body, for example, it can be applied to mammals including humans.

[0032] The cleaning composition of the present invention exhibits a cleaning effect even with two components, sophorolipids and SDS, but may further contain other cleaning components or components generally formulated in cleaning compositions depending on the object to be cleaned. Other cleaning components are known, and one or more cleaning components (e.g., surfactants, alkalis, abrasives, enzyme preparations, bleaching agents, disinfectants, deodorants) or other optional components (e.g., solvents, dispersants, pH adjusters, antioxidants, fragrances, colorants, viscosity adjusters, preservatives) can be formulated according to the cleaning purpose. For example, when the cleaning composition of the present invention is provided in the form of a medical instrument cleaner, it can be provided as an alkaline cleaner containing an alkali such as potassium hydroxide or sodium hydroxide in addition to sophorolipids and SDS, or it can be provided as an enzyme cleaner containing a proteolytic enzyme and a surfactant in addition to sophorolipids and SDS. Also, when the cleaning composition of the present invention is provided in the form of a disinfectant or a bleaching agent, a disinfectant or a bleaching agent (e.g., chlorine-based bleaching agent, enzyme-based bleaching agent) can be formulated in addition to sophorolipids and SDS.

[0033] In its usage mode, the cleaning composition of the present invention can be used so as to have a specific sophorolipid concentration. The lower limit value of the sophorolipid concentration in this case can be, for example, 0.00001% by mass, 0.0001% by mass or 0.001% by mass, and the upper limit value can be, for example, 1% by mass, 0.1% by mass or 0.01% by mass. These lower limit values and upper limit values can be arbitrarily combined respectively, and the range of the sophorolipid concentration can be, for example, 0.00001 - 1% by mass or 0.0001 - 0.1% by mass. In its usage mode, the cleaning composition of the present invention can also be used so as to have a specific SDS concentration. The lower limit value of the SDS concentration in this case can be, for example, 0.0001% by mass, 0.001% by mass or 0.01% by mass, and the upper limit value can be, for example, 1% by mass or 0.1% by mass. These lower limit values and upper limit values can be arbitrarily combined respectively, and the range of the SDS concentration can be, for example, 0.0001 - 1% by mass or 0.001 - 0.1% by mass.

[0034] In the cleaning composition of the present invention, the contents of sophorolipid and SDS can be arbitrarily set according to the usage form of the cleaning composition by referring to the above-mentioned usage concentrations. That is, the cleaning composition of the present invention can determine the contents so that sophorolipid and SDS are at effective concentrations in the assumed usage form. When the cleaning composition of the present invention is provided in the form of a liquid medical instrument cleaner (assuming 200-fold dilution during use), for example, the concentration of sophorolipid can be 0.002 to 20% by mass, and the concentration of SDS can be 0.02 to 20% by mass.

[0035] <<Antifouling composition>> Further, the antifouling composition of the present invention can be provided as an antifouling composition for a target substrate in which the formation of biofilm is not desired, including the above-mentioned target substrate. Specifically, when the antifouling composition of the present invention is for the purpose of antifouling of objects such as medical instruments, water treatment facilities, air conditioning facilities, ship hulls and port facilities, fishing gear, etc., for which antifouling is required, it can be provided as an antifouling paint or an antifouling coating agent.

[0036] The antifouling composition of the present invention exhibits an antifouling effect even with two components, sophorolipid and SDS, but may further contain other antifouling components according to the antifouling target and components generally blended in the antifouling composition. Other antifouling components are known, and one or more antifouling components (for example, cuprous oxide, zinc oxide, organic nitrogen sulfur-based compounds, organic boron-based compounds) and other optional components (for example, solvents, paint components, coating components) can be blended according to the antifouling purpose.

[0037] In its usage mode, the antifouling composition of the present invention can be used to achieve a specific concentration of sophorolipid. In this case, the lower limit of the sophorolipid concentration can be, for example, 0.00001% by mass, 0.0001% by mass or 0.001% by mass, and the upper limit can be, for example, 1% by mass, 0.1% by mass or 0.01% by mass. These lower and upper limits can be arbitrarily combined respectively, and the above range of sophorolipid concentration can be, for example, 0.00001 - 1% by mass or 0.0001 - 0.1% by mass. The antifouling composition of the present invention can also be used to achieve a specific SDS concentration in its usage mode. In this case, the lower limit of the SDS concentration can be, for example, 0.0001% by mass, 0.001% by mass or 0.01% by mass, and the upper limit can be, for example, 1% by mass or 0.1% by mass. These lower and upper limits can be arbitrarily combined respectively, and the above range of SDS concentration can be, for example, 0.0001 - 1% by mass or 0.001 - 0.1% by mass.

[0038] The contents of sophorolipid and SDS in the antifouling composition of the present invention can be arbitrarily set according to the usage form of the antifouling composition by referring to the usage concentrations as described above. That is, the antifouling composition of the present invention can determine the contents so that sophorolipid and SDS are at effective concentrations in the assumed usage form.

[0039] <<Virus Inactivation Composition>> The virus inactivation composition of the present invention can be provided as a composition for disinfecting a target substrate on which a virus is present and adheres, including the above-mentioned target substrate. Specifically, when the virus inactivation composition of the present invention is for the purpose of disinfecting medical instruments, it can be used as a medical instrument virus disinfectant; when it is for the purpose of disinfecting kitchen utensils, it can be used as a kitchen virus disinfectant; when it is for the purpose of disinfecting the skin, it can be used as soap, hand soap, or body soap; when it is for the purpose of disinfecting the oral cavity, it can be used as an oral virus disinfectant (for example, mouthwash); when it is for the purpose of disinfecting dentures and removable teeth, it can be used as a denture virus disinfectant and a removable tooth virus disinfectant; when it is for the purpose of disinfecting the toilet, it can be used as a toilet virus disinfectant, respectively. Since various problems occur due to virus adhesion, the virus inactivation composition of the above aspect is expected to remove or suppress the formation of biofilms from the target substrate and inactivate the viruses adhering to the target substrate. In addition, when applied to a living body, for example, it can be applied to mammals including humans. When the virus inactivation composition of the present invention is also intended for cleaning, antifouling, etc. in addition to disinfecting the target substrate, it can also be used as a virus disinfecting cleaner or a virus disinfecting antifouling agent.

[0040] The virus inactivation composition of the present invention exhibits a virus inactivation effect even with two components, sophorolipid and SDS, but may further contain other antiviral components according to the disinfection target and components generally formulated in the virus inactivation composition. Other antiviral components are known, and one or more antiviral components (for example, ionic surfactants such as benzalkonium chloride) according to the disinfection purpose, the above-mentioned cleaning components, and other optional components can be formulated.

[0041] In the usage mode of the virus inactivation composition of the present invention, it can be used to achieve a specific concentration of sophorolipid. In this case, the lower limit of the sophorolipid concentration can be, for example, 0.00001% by mass, 0.0001% by mass or 0.001% by mass, and the upper limit can be, for example, 1% by mass, 0.1% by mass or 0.01% by mass. These lower and upper limits can be arbitrarily combined respectively, and the range of the above-mentioned sophorolipid concentration can be, for example, 0.00001 - 1% by mass or 0.0001 - 0.1% by mass. The virus inactivation composition of the present invention can also be used to achieve a specific SDS concentration in its usage mode. In this case, the lower limit of the SDS concentration can be, for example, 0.0001% by mass, 0.001% by mass or 0.01% by mass, and the upper limit can be, for example, 1% by mass or 0.1% by mass. These lower and upper limits can be arbitrarily combined respectively, and the range of the above-mentioned SDS concentration can be, for example, 0.0001 - 1% by mass or 0.001 - 0.1% by mass.

[0042] The contents of sophorolipid and SDS in the virus inactivation composition of the present invention can be arbitrarily set according to the usage form of the virus inactivation composition by referring to the usage concentrations as described above. That is, the virus inactivation composition of the present invention can determine the contents so that sophorolipid and SDS are at effective concentrations in the assumed usage form.

[0043] The disinfection composition, cleaning composition, antifouling composition and virus inactivation composition of the present invention can be provided as a liquid composition according to their usage modes, but can also be provided as a solid composition or powder composition and dissolved in a solvent such as water for use during use.

[0044] In the disinfectant composition, cleaning composition, antifouling composition, and virus inactivation composition of the present invention, even at low concentrations of sophorolipids and SDS, they exhibit a disinfection effect and a cleaning effect, and are particularly effective in removing and suppressing the formation of biofilms and inactivating viruses. Therefore, according to the disinfectant composition, cleaning composition, antifouling composition, and virus inactivation composition of the present invention, the amount of expensive sophorolipids used can be reduced, and the amount of non-natural surfactants used can also be reduced, providing a disinfectant composition, cleaning composition, antifouling composition, and virus inactivation composition that are low-cost and environmentally friendly.

[0045] <<Composition for Removing and Suppressing Formation of Biofilm>> According to another aspect of the present invention, there are provided a composition and an agent for removing biofilm containing sophorolipids and SDS as active ingredients. In the present invention, "removing biofilm" means removing the biofilm adhering to the target substrate. Biofilm is a form of dirt, and removing biofilm is a form of cleaning.

[0046] For the removal of biofilm, for example, as described in Example 2 below, when the test sample is brought into contact with the biofilm formed on the surface of the object by culturing bacteria for a certain period of time and when it is not brought into contact (control), there are methods such as comparing the coverage state of the biofilm at the start of contact with the coverage state of the biofilm after a certain period of time has elapsed since the start of contact. In this case, if the amount of biofilm coverage when the test sample is brought into contact is reduced compared to the control, it can be evaluated that the test sample has an effect of removing biofilm.

[0047] The composition for removing biofilm of the present invention can be implemented in the form of the above-described cleaning composition of the present invention.

[0048] According to another aspect of the present invention, there are also provided a composition for inhibiting biofilm formation and an inhibitor, which comprise sophorolipid and SDS as active ingredients. In the present invention, "inhibiting biofilm formation" means inhibiting the adhesion of biofilm to a target substrate, and is used in the sense of including inhibiting the adhesion of bacteria forming biofilm to the target substrate. Biofilm is a form of dirt, and inhibiting biofilm formation is a form of antifouling.

[0049] For inhibiting biofilm formation, for example, as described in Example 5 below, bacteria are inoculated respectively when a test sample is brought into contact with the surface of a target substrate and when it is not brought into contact (control), and the adhesion state of bacteria at the start of inoculation and the adhesion state of bacteria after a certain period of time has elapsed since inoculation are compared. In this case, if the amount of bacteria adhered after a certain period of time when the test sample is brought into contact is suppressed compared to the control, it can be evaluated that the test sample has an effect of inhibiting biofilm formation.

[0050] The composition for inhibiting biofilm formation of the present invention can be implemented in the form of the above-described cleaning composition and antifouling composition of the present invention.

[0051] <<Non-bactericidal composition>> According to another aspect of the present invention, there is also provided a non-bactericidal composition for removing and / or inhibiting the formation of biofilm, which comprises sophorolipid. Since sophorolipid has an effect of removing biofilm or inhibiting its formation without killing bacteria as described in the examples below, it can be used in a form expected to suppress or avoid the generation of resistant bacteria. Specifically, the non-bactericidal composition of the present invention can be applied to an article for which the generation of resistant bacteria should be suppressed, and can also be applied in an environment for which the generation of resistant bacteria should be suppressed.

[0052] Examples of articles to which the non-bactericidal composition of the present invention should be applied to suppress the generation of resistant bacteria include those that may pose a risk to the human body and the environment, such as medical devices and ship bottoms. Examples of environments to which the non-bactericidal composition of the present invention should be applied to suppress the generation of resistant bacteria include places where there is concern about the impact on the biological system, such as the sea, rivers, lakes, and the like.

[0053] In the usage mode of the non-bactericidal composition of the present invention, it can be used so as to achieve a specific concentration of sophorolipid. In this case, the lower limit value of the sophorolipid concentration can be, for example, 0.01% by mass or 0.1% by mass, and the upper limit value can be, for example, 2% by mass or 1% by mass. These lower limit values and upper limit values can be arbitrarily combined, and the range of the sophorolipid concentration can be, for example, 0.01 to 2% by mass or 0.1 to 1% by mass.

[0054] The non-bactericidal composition of the present invention can be implemented in the form of the above-described cleaning composition or antifouling composition of the present invention.

[0055] <<Methods and Uses>> According to another aspect of the present invention, there are provided a disinfection method, a cleaning method, an antifouling method, and a virus inactivation method characterized by using sophorolipid and SDS. The disinfection method, cleaning method, antifouling method, and virus inactivation method of the present invention can be implemented by applying or bringing sophorolipid and SDS into contact with the target substrate. For example, sophorolipid and SDS can be added to, applied to, sprayed on, or dispersed on the target substrate, or the target substrate can be immersed in a mixture of sophorolipid and SDS. The target substrate is as described above, and the disinfection method, cleaning method, antifouling method, and virus inactivation method of the present invention can be implemented with respect to the target substrate.

[0056] In the disinfection method, cleaning method, anti-fouling method, and virus inactivation method of the present invention, as long as the effects are achieved, the contact order and contact timing of sophorolipids and SDS are not limited. Sophorolipids and SDS may be simultaneously contacted with the object to be contacted, or sophorolipids and SDS may be contacted at separate timings. In a preferred embodiment of the disinfection method, cleaning method, anti-fouling method, and virus inactivation method of the present invention, sophorolipids and SDS can be simultaneously contacted with the object to be contacted. In this case, sophorolipids and SDS may be separate preparations or a mixture of sophorolipids and SDS such as the composition of the present invention.

[0057] In the disinfection method, cleaning method, anti-fouling method, and virus inactivation method of the present invention, sophorolipids and SDS can be used at specific concentrations with respect to the target substrate, and the concentrations of sophorolipids and SDS can be set in the same manner as described in the disinfection composition, cleaning composition, anti-fouling composition, and virus inactivation composition of the present invention. In the disinfection method, cleaning method, anti-fouling method, and virus inactivation method of the present invention, for example, sophorolipids can be used at a concentration of 0.00001 to 1% by mass or 0.0001 to 0.1% by mass, and SDS can be used at a concentration of 0.0001 to 1% by mass or 0.001 to 0.1% by mass.

[0058] According to still another aspect of the present invention, there is provided a biofilm removal method characterized by using sophorolipids and SDS. The biofilm removal method of the present invention can be carried out by applying or contacting sophorolipids and SDS to the biofilm. For example, sophorolipids and SDS can be added, applied, sprayed, or scattered onto the target substrate, or the target substrate can be immersed in a mixture of sophorolipids and SDS. The target substrate on which biofilms may be formed is as described above, and the biofilm removal method of the present invention can be carried out on the target substrate.

[0059] In the biofilm removal method of the present invention, sophorolipid and SDS can be used for the target substrate so as to have specific concentrations respectively, and the concentrations of sophorolipid and SDS can be set in the same manner as described in the cleaning composition of the present invention. In the biofilm removal method of the present invention, for example, sophorolipid can be used at a concentration of 0.00001 to 1% by mass or 0.0001 to 0.1% by mass, and SDS can be used at a concentration of 0.0001 to 1% by mass or 0.001 to 0.1% by mass.

[0060] According to still another aspect of the present invention, there is also provided a biofilm formation inhibition method characterized by using sophorolipid and SDS. The biofilm formation inhibition method of the present invention can be carried out by applying or bringing sophorolipid and SDS into contact with a target substrate on which biofilm can be formed. For example, sophorolipid and SDS can be added to, applied to, sprayed or scattered on the target substrate, or the target substrate can be immersed in a mixture of sophorolipid and SDS. The target substrate on which biofilm may be formed is as described above, and the biofilm formation inhibition method of the present invention can be carried out on the said target substrate.

[0061] In the biofilm formation inhibition method of the present invention, sophorolipid and SDS can be used for the target substrate so as to have specific concentrations respectively, and the concentrations of sophorolipid and SDS can be set in the same manner as described in the antifouling composition of the present invention. In the biofilm formation inhibition method of the present invention, for example, sophorolipid can be used at a concentration of 0.00001 to 1% by mass or 0.0001 to 0.1% by mass, and SDS can be used at a concentration of 0.0001 to 1% by mass or 0.001 to 0.1% by mass.

[0062] In the biofilm removal method and biofilm formation inhibition method of the present invention, as long as the biofilm removal effect and biofilm formation inhibition effect are achieved, the contact order and contact timing of sophorolipid and SDS are not limited. Whether the sophorolipid and SDS are simultaneously contacted with the contact target or the sophorolipid and SDS are contacted at separate timings is acceptable. In a preferred embodiment of the biofilm removal method and biofilm formation inhibition method of the present invention, the sophorolipid and SDS can be simultaneously contacted with the contact target. In this case, the sophorolipid and SDS may be separate preparations or a mixture of sophorolipid and SDS such as the cleaning composition of the present invention or the antifouling composition of the present invention.

[0063] The contact method in the biofilm removal method and biofilm formation inhibition method of the present invention is not particularly limited as long as the biofilm removal effect and biofilm formation inhibition effect are achieved. For example, it can be carried out by addition, coating, spraying, scattering, or immersion.

[0064] According to another aspect of the present invention, there is provided a method for removing and / or inhibiting the formation of biofilms, characterized by using sophorolipid. The biofilm removal and / or formation inhibition method of the present invention is characterized by not killing bacteria, that is, being non-bactericidal. The biofilm removal and / or formation inhibition method of the present invention can also be carried out by applying or contacting the sophorolipid to the target substrate. For example, the sophorolipid can be added, coated, sprayed, or scattered on the target substrate, or the target substrate can be immersed in a sophorolipid solution. The target substrate is as described above, and the biofilm removal and / or formation inhibition method of the present invention can be carried out on the target substrate.

[0065] In the method for removing and / or suppressing the formation of a biofilm of the present invention, sophorolipid can be used at a specific concentration with respect to a target substrate, and can be set in the same manner as described in the non-bactericidal composition of the present invention. In the method for removing and / or suppressing the formation of a biofilm of the present invention, for example, sophorolipid can be used at a concentration of 0.01 to 2% by mass or 0.1 to 1% by mass.

[0066] According to another aspect of the present invention, there is provided the use of a combination of sophorolipid and SDS for use in disinfection, cleaning, antifouling, virus inactivation, biofilm removal and / or biofilm formation suppression, or as a disinfectant, cleaning agent, antifouling agent, virus disinfectant, biofilm removal agent and / or biofilm formation suppression agent. According to the present invention, there is also provided the use of a combination of sophorolipid and SDS for the manufacture of a disinfectant, cleaning agent, antifouling agent, virus disinfectant, biofilm removal agent and / or biofilm formation suppression agent. The above use of the present invention can be carried out according to the descriptions of the disinfection composition, cleaning composition, antifouling composition and virus inactivation composition of the present invention and the disinfection method, cleaning method, antifouling method and virus inactivation method of the present invention.

[0067] According to another aspect of the present invention, there is provided the non-bactericidal use of sophorolipid for use in biofilm removal and / or biofilm formation suppression, or as a biofilm removal agent and / or biofilm formation suppression agent. According to the present invention, there is also provided the use of sophorolipid for the manufacture of a non-bactericidal biofilm removal agent and / or non-bactericidal biofilm formation suppression agent. The above use of the present invention can be carried out according to the description of the non-bactericidal composition of the present invention.

[0068] According to the present invention, the following inventions are provided.

[0101] A cleaning composition comprising sophorolipid and sodium dodecyl sulfate (SDS).

[0102] An antifouling composition comprising sophorolipid and sodium dodecyl sulfate (SDS).

[0103] The composition according to the above

[0101] or

[0102] for use in removing and / or suppressing the formation of biofilms.

[0104] The composition according to the above

[0103] , wherein the biofilm is derived from Gram-negative bacteria.

[0105] The composition according to any one of the above

[0101] to

[0104] , wherein sophorolipid is used at a concentration of 0.1% by mass or less.

[0106] The composition according to any one of the above

[0101] to

[0105] , wherein SDS is used at a concentration of 0.1% by mass or less.

[0107] A composition for removing and / or suppressing the formation of biofilms, comprising sophorolipid and sodium dodecyl sulfate (SDS).

[0108] A method for removing and / or suppressing the formation of biofilms, characterized by using sophorolipid and sodium dodecyl sulfate (SDS).

[0109] The method according to the above

[0108] , wherein the biofilm is derived from Gram-negative bacteria.

[0110] The method according to the above

[0108] or

[0109] , characterized by using the composition according to any one of the above

[0101] to

[0107] as sophorolipid and SDS.

[0111] The method according to any one of the above

[0108] to

[0110] , wherein sophorolipid is used at a concentration of 0.1% by mass or less.

[0112] The method according to any one of the above

[0108] to

[0111] , wherein SDS is used at a concentration of 0.1% by mass or less.

Examples

[0069] The present invention will be described more specifically based on the following examples, but the present invention is not limited to these examples.

[0070] Example 1: Distribution of surface tension and micelle diameter of various surfactants (1) Method a Surfactant As various surfactants, sophorolipids (Aralide Carbon Solutions, ratio of lactone type to acidic type is 7:3, pH is 6.5 - 7.0), SDS (Nacalai Tesque), tween20 (Nacalai Tesque) and tween80 (Nacalai Tesque) were used. Unless otherwise specified, the same surfactants as those described in this paragraph were used in other examples as well.

[0071] I Measurement of surface tension and micelle size distribution Surface tension was measured by dissolving various surfactants in deionized water and measuring the values at each concentration at 25.2 °C using an automatic surface tensiometer (Kyowa Interface Science). Also, the micelle size distribution was measured by dynamic light scattering using a Zetasizer Nano ZS (Malvern Panalytical). Unless otherwise specified, the measurements were carried out in the same manner as described in this paragraph in other examples.

[0072] (2) Results The results were as shown in Figure 2. Since the critical micelle concentration (CMC) determines the activity or surface property of the surfactant, the characteristics of surface tension (γ) and micelle diameter were confirmed as the functions of various surfactants. Based on the CMC measured here, the concentration ranges of various surfactants to be examined were determined.

[0073] Example 2: Biofilm removal effect by various surface active substances (1) Method A Microfluidics The master of the microfluidic chip was spin-coated with a negative photoresist KMPR (MicroChem) on a silicon nitride wafer (Mechanical grade, UniversityWafer) and exposed through a photomask using a mask aligner UV-KUB2 (Kloe). The photoresist was developed to obtain the master mold.

[0074] The master replication was performed using a polydimethylsiloxane (PDMS) Sylgard 184 silicone elastomer kit (Toray Dow Corning). To prepare the PDMS, the prepolymer and crosslinking reagent were mixed at a ratio of 10:1, poured into the master mold, degassed under vacuum to remove trapped air bubbles, and then 2 bonded to the coverslip using a plasma cleaner (CUTE, Femto-Science). The height of the channels was measured using a 3D laser scanning confocal microscope VK-X160K (Keyence). The dimensional channels had a height of 70 μm and a width of 200 μm.

[0075] The microfluidic channels were designed as shown in Figure 3. Specifically, it consisted of five inlets and one outlet, including a dedicated port (inlet (i)) for injecting the inoculum and four inlets (ii, iii, iv, and v) capable of injecting different solutions simultaneously, and an outlet (vi) for drainage.

[0076] E. coli cells The cells used were Pseudomonas aeruginosa ( Pseudomonas aeruginosa , strain: PAO1, available from the RIKEN BioResource Center (BRC)). Pseudomonas aeruginosa is a widely distributed Gram-negative bacterium that exhibits swimming and surface motility. PAO1 secretes multiple polysaccharides that help firmly attach biofilm communities to the surface while providing internal structural support. PAO1 was selected due to its ubiquity, its ability to form high-density biofilms in porous environments that strongly impede its flow, and its status as an important human pathogen.

[0077] G. Bacterial culture OD 600nm PAO1 was cultured in 4 mL of LB medium overnight at 37 °C and 190 rpm in an orbital shaker until the mid-logarithmic phase with an OD = 0.5. Next, this culture solution was added to an LB-modified medium (which was obtained by diluting the carbon source concentration in the LB medium to one-tenth while maintaining the same NaCl concentration as in the LB medium) (the addition amount was such that it did not affect the composition of the LB-modified medium), and OD600nm It was diluted so that it became 0.1. Subsequently, 100 μL of this subculture solution was inoculated into the inlet (i) of the microfluidics through a silicon tube connected to a 1 mL syringe. In this example, except for Example 7, OD 600nm was measured using MP-1200 (ERMA).

[0078] A positive syringe pump (Harvard Apparatus) was used to simultaneously draw out into four separate syringes at inlets (ii, iii, iv, and v). The microfluidic device was heated to 30 °C with a microscope stage heater (Tokai Hit).

[0079] E Surfactant After dissolving various surfactants in deionized water containing 10% by mass DMSO, they were diluted with the LB modified medium so that the final concentration of DMSO was 1% by mass or less, which had no effect on cell growth, and then used.

[0080] O Flushing test First, as described in U above, to grow the biofilm of PAO1, PAO1 was inoculated into all channels through inlet (i) to attach the cells. After cell attachment, fresh medium as food was injected into the four flow channels simultaneously using a syringe pump and incubated for 12 hours.

[0081] Next, each surfactant prepared in E was injected into the channels, and the change in the surface coating area of the biofilm was measured. In this test, when the medium containing the surfactant was flowed, time-lapse images were recorded every minute to track the change in surface coating. Specifically, a time-lapse imaging Axio Observer Z1 inverted microscope (Carl Zeiss) with a camera (Andor) was used to monitor the surface coating formed in the channels by recording images from each channel every minute. Then, image processing was performed using ImageJ, the cells were thresholded and segmented, and the built-in function was used to calculate the total surface coating (A). At the start of the measurement (time 0), the channels were completely covered with the biofilm, and the total surface coating at this time was taken as A 0 and.

[0082] (2) Results The results were as shown in Fig. 4. Specifically, it was confirmed that the biofilm removal effect from the channels was superior for SLx compared to SDS, tween20, and tween80. For example, when the concentration of SLx was 0.1% by mass, more than 50% of the mature biofilm was removed after 5 hours. Also, when the concentration of SLx was increased up to 1% by mass, the removal rate of the biofilm increased, and as shown in Fig. 4, 50% of the biofilm was removed in 2 hours and more than 99% was removed in just 4 hours. Also, the images after 5 hours of surfactant treatment were as shown in Fig. 5.

[0083] (3) Discussion The results in (2) above indicate that SLx has a strong removal effect on biofilms, that SDS also has a certain degree of removal effect, while Tween has low removal ability. However, the drawback of bio-based surfactants such as SLx is that their production costs are high. Therefore, if the usage amounts of these can be reduced, they have the potential to be useful biofilm removal agents. As these results show, the removal effect of SLx is concentration-dependent. For this reason, further investigation was carried out to see if the biofilm removal effect could be exerted at a lower concentration.

[0084] Example 3: Influence on surface tension of the combination of sophorolipid and SDS Prior to examining whether the biofilm-removing effect can be exhibited at a lower concentration for SLx as described in Example 2(3), in Example 3, SLx and SDS were mixed at low concentrations and the surface tension was measured. As a result, it was as shown in Table 1. When SLx and SDS were added alone at their respective concentrations, there was no effect on the surface tension. However, when a mixed solution of SLx and SDS was added, it was confirmed that the surface tension decreased.

[0085]

Table 1

[0086] Example 4: Biofilm removal effect by the combination of sophorolipid and SDS Based on the results examined in Example 3, the biofilm-removing effect by the rinsing test at SLx concentrations (0.001 mass%, 0.01 mass%) and SDS concentrations (0.01 mass%, 0.1 mass%) was further examined.

[0087] It was carried out in the same manner as described in Example 2. As a result, it was as shown in Fig. 6. When SLx was added alone at a concentration of 0.01% by mass, more than 50% of the biofilm was removed 5 hours after the addition. On the other hand, when SDS was added at a concentration of 0.1% by mass to this, almost 80% of the biofilm was removed 2 hours later, and the same removal effect as that after 5 hours in the case of 0.1% by mass was exhibited in the case of 0.01% by mass (upper part of Fig. 6). Also, when SLx was added alone at a concentration of 0.001% by mass, the change in the surface coating of the biofilm was at an unmeasurable level compared to the negative control (0% by mass). However, when combined with the above concentration of SDS, it was confirmed that even this low concentration of SLx was effective in removing the biofilm (lower part of Fig. 6). Also, even at a concentration where no effect of removing the biofilm was observed when SDS was used alone (Fig. 4, SDS concentration: 0.01% by mass), the effect of removing the biofilm was exhibited by combining with SLx, and thus the effect of removing the biofilm by the combination of SLx and SDS was shown.

[0088] Regarding the above results, the concentrations of SLx and SDS were normalized by their respective CMCs, and the results plotted as a "score" representing the ability of the mixed solution to remove the biofilm are shown in Fig. 7. The said "score" was calculated as "1 - A / A0", and the score becomes 1 when the biofilm is completely removed after 5 hours. From this figure too, the effect of removing the biofilm by the combination of SLx and SDS can be confirmed.

[0089] On the other hand, the effect of removing the biofilm by the combination of SLx and Tween 20, Tween 80 was also examined. The results were as shown in Fig. 8. It was confirmed that the combination of SLx with any of the Tweens had almost no effect on removing the biofilm.

[0090] Also, the effect of removing the biofilm by the combination of SDS and Tween 20, Tween 80 was as shown in Fig. 9. It was confirmed that the combination of SDS and Tween 80 also had an effect on removing the biofilm.

[0091] A problem with biosurfactants is that their production cost is higher than that of synthetic surfactants. Therefore, it would be a great advantage if the ability to remove biofilms could be enhanced while reducing the total amount used. The present invention enables the removal effect of mature biofilms to be enhanced in a shorter time and at a lower concentration. It is also expected to lead to a reduction in the pollution of the environment and biological systems by surfactants.

[0092] Example 5: Cell adhesion inhibitory effect by the combination of sophorolipid and SDS (1) Method a Microfluidics The microfluidics described in Example 2(1)a was used.

[0093] b Bacterial cells The bacterial cells described in Example 2(1)b were used.

[0094] c Bacterial culture OD 600nm PAO1 was cultured in 4 mL of LB medium overnight at 37°C and 190 rpm in an orbital shaker until the middle of the logarithmic phase with OD 600nm = 0.5. Next, this culture solution was added to an LB-modified medium (a medium in which the concentration of the carbon source in the LB medium was diluted to 1 / 10 and the NaCl concentration was the same as that of the LB medium) (the addition amount was such that it did not affect the composition of the LB-modified medium), and diluted to OD

[0095] A positive syringe pump (Harvard Apparatus) was used to simultaneously draw 100 μL of this subculture solution into the inlets (i), (ii), (iii), (iv), and (v) through a silicon tube connected to a 1 mL syringe. The microfluidic device was heated to 30°C with a microscope stage heater (Tokai Hit).

[0096] d Surfactant Example 2 (1) Various surfactants were diluted with the LB-modified medium and used in the same manner as described in E.

[0097] O Cell adhesion inhibition test Regarding the cell adhesion inhibition effect under the condition of injection at a flow rate (Q) of 100 μL / hour with each combination of SLx (0.001% by mass) and SDS (0.1% by mass), Tween 20 (0.1% by mass), and Tween 80 (0.1% by mass) prepared in E, it was confirmed by measuring the change over time in the covered area (A) by the attached cells. The measurement start point (time 0) was set to 1 hour after the above subculture solution inoculation. Specifically, a time-lapse imaging Axio Observer Z1 inverted microscope (Carl Zeiss) equipped with a camera (Andor) was used, and the surface coverage of the attached cells was monitored by recording the images of the channels. The bright-field images were processed by ImageJ, and after subtracting the background, the images were threshold-processed to segment single cells. Then, the total surface coverage area of the cells was calculated, and the surface coverage area (%) was calculated by dividing it by the entire visible area.

[0098] (2) Results The results were as shown in Fig. 10. When SLx and SDS were combined, it was confirmed that the surface coverage after 8 hours was 0, indicating a significant cell adhesion inhibition effect. Also, when SLx and Tween 80 were combined, it was confirmed that the surface coverage after 8 hours was about 10%, showing a cell adhesion inhibition effect. On the other hand, when SLx was added alone and when SLx and Tween 20 were combined, it was confirmed that the surface coverage after 8 hours reached nearly 80%, indicating a poor cell adhesion inhibition effect.

[0099] Example 6: Bacteriostatic effect and bactericidal effect by various surfactants (1) Method A Bacterial cells The bacterial cells described in Example 2 (1) ii were used.

[0100] B Surfactants Various surfactants were diluted with the LB-modified medium and used in the same manner as described in Example 2 (1) E.

[0101] Evaluation of bacteriostatic effect Perform bacterial culture in the same manner as described in Example 2(1), add various surfactants at different concentrations, culture at 37 °C and 190 rpm on an orbital shaker, and evaluate the bacteriostatic effect by measuring the absorbance (OD600 nm ) as a function of time for bacterial growth.

[0102] Evaluation of bactericidal effect OD600 nm After culturing in 4 mL of LB medium until OD600 reaches 0.15, add various surfactants at different concentrations and culture at 37 °C and 190 rpm on an orbital shaker. Then, measure the absorbance (OD600 nm ) in the mid-exponential phase. Also, although absorbance is useful for measuring the number of cells in a solution, information regarding cell viability cannot be obtained, so colony-forming units (CFU) were also calculated. Specifically, after directly adding SLx to the culture solution and culturing for 12 hours, the culture was serially diluted with fresh LB, and immediately 100 μL of the suspension was evenly spread on an LB agar plate to seed the cells and cultured overnight. Then, the number of colonies on the plate where the colonies were well separated was counted, and CFU / mL was calculated.

[0103] (2) Results The results of the bacteriostatic effect were as shown in Fig. 11A. As a result of measuring the absorbance (OD 600 ) in liquid culture containing various surfactants at different concentrations in LB medium and evaluating cell growth, no inhibitory effect on bacterial growth was observed for SLx, Tween 20, and Tween 80. In contrast, it was confirmed that SDS inhibits bacterial growth as the concentration increases, and at 1% by mass, bacterial growth was greatly suppressed, but after about 10 hours, OD 600 showed a gradually increasing trend.

[0104] The results of the bactericidal effect were as shown in Figs. 11B and 11C. In Fig. 11B, no inhibition of bacterial growth was observed for SLx, Tween 20, and Tween 80, whereas for SDS at 1% by mass, OD decreased in the first 3 hours600 caused a decrease, and OD began to increase 5 hours after addition. 600 It was confirmed that. OD 600 The first decrease in was due to cell lysis immediately after the addition of SDS, and the increase in OD about 5 hours later 600 was considered to indicate recovery and regrowth. In Figure 11C, from the values of OD 600 and CFU / mL, it was found that SLx had no effect on the survival of bacteria, that is, no bactericidal effect was observed with SLx.

[0105] Example 7: Bacteriostatic effect by the combination of sophorolipid and SDS In Example 7, the bacteriostatic effect of the combination of sophorolipid and SDS was investigated.

[0106] (1) Method a Bacterial cells The bacterial cells described in Example 2(1)i were used.

[0107] b Surfactants The same various surfactants as described in Example 2(1)iii were used.

[0108] c Evaluation of bacteriostatic effect OD 600nm PAO1 was cultured in 4 mL of LB medium overnight at 37 °C and 190 rpm in an orbital shaker until the middle of the logarithmic phase with OD = 0.5. Next, this culture solution was added to LB modified medium (the concentration of the carbon source in LB medium was diluted to 1 / 10, and the NaCl concentration was the same as that of LB medium) (the addition amount was such that it did not affect the composition of LB modified medium), and diluted to OD 600nm = 0.01. Subsequently, it was seeded in a 96-well plate, various surfactants were added at each concentration, and then cultured at 37 °C and 425 rpm in a double orbital shaker for 12 hours. Then, the bacteriostatic effect was evaluated by measuring the absorbance (OD 600nm ) as a function of time. In Example 7, OD 600nm was measured using Cytation 5 (BioTek).

[0109] (2) Results The results were as shown in Fig. 12. No bacteriostatic effect was observed with the combination of sophorolipid and SDS.

[0110] Example 8: Inactivation of virus by sophorolipid (1) Method As the virus, SARS-CoV-2, the causative virus of COVID-19 (obtained from the National Institute of Infectious Diseases), was used (virus titer: 3×10 7 TCID 50 / ml). The virus sample was diluted 10-fold with each concentration of SLx, incubated at room temperature for 5 minutes, and then the surfactant (sophorolipid) was neutralized with 10 volumes of SCDLP culture medium. Subsequently, this virus solution was diluted 4-fold with DMEM medium and used to infect human Vero-E6-TMPRSS2 cells (obtained from the National Institute of Biomedical Innovation, Health and Nutrition) in a 96-well plate (n = 2). The number of wells showing the cytopathic effect induced by SARS-CoV-2 48 hours after infection was counted, and the virus titer (infection rate) was measured.

[0111] (2) Results The results were as shown in Fig. 13. Specifically, when the virus titer of the control (without SLx) was set as the infection rate of 100%, the infection rate of the samples pretreated with SLx was suppressed to several percent at any concentration. From this result, it was confirmed that SLx has an effect of inactivating the virus.

[0112] Example 9: Virus inactivation by the combination of sophorolipid and SDS In Example 9, virus inactivation by the combination of sophorolipid and SDS was examined.

[0113] (1) Method SARS-CoV-2 was prepared from the culture solution collected from infected human Vero-E6-TMPRSS2 cells (virus titer: 3×10 7 TCID 50 / ml), and the same method as described in Example 8(1) was performed except that SLx and SDS were used as surfactants.

[0114] (2) Results The results were as shown in Table 2 and Fig. 14. Specifically, it was confirmed that virus infection was further suppressed when treated with a combination of SLx and SDS, as compared with the case of treatment with SLx alone. On the other hand, when treated with SDS alone, no effect of suppressing infection was confirmed.

[0115]

Table 2

Claims

1. A disinfectant composition comprising sophorolipid and sodium dodecyl sulfate (SDS), wherein the concentration of the sophorolipid is 0.001% by mass or more and 0.1% by mass or less, and the concentration of the SDS is 0.001% by mass or more and 0.01% by mass or less.

2. A cleaning composition or an antifouling composition comprising sophorolipid and sodium dodecyl sulfate (SDS), wherein the concentration of the sophorolipid is 0.001% by mass or more and 0.01% by mass or less, and the concentration of the SDS is 0.001% by mass or more and 0.01% by mass or less.

3. A virus-inactivating composition comprising sophorolipid and sodium dodecyl sulfate (SDS), wherein the virus is SARS-CoV-2, the concentration of the sophorolipid is 0.001% by mass or more and 0.1% by mass or less, and the concentration of the SDS is 0.001% by mass or more and 0.1% by mass or less.

4. The composition according to any one of claims 1 to 3, for use in removing and / or suppressing the formation of biofilms.

5. The composition according to claim 4, wherein the biofilm is derived from Gram-negative bacteria.

6. A composition for removing and / or suppressing the formation of biofilms, comprising sophorolipid and sodium dodecyl sulfate (SDS), wherein the concentration of the sophorolipid is 0.001% by mass or more and 0.01% by mass or less, and the concentration of the SDS is 0.001% by mass or more and 0.01% by mass or less.

7. A disinfection method characterized by using sophorolipid and sodium dodecyl sulfate (SDS), wherein the sophorolipid is used at a concentration of 0.1% by mass or less, and the SDS is used at a concentration of 0.01% by mass or less (provided that in the method, humans are excluded from the target substrate to which the sophorolipid and the SDS are applied).

8. A cleaning method, an antifouling method, and a method for removing and / or suppressing the formation of biofilms, characterized by using sophorolipid and sodium dodecyl sulfate (SDS), wherein the sophorolipid is used at a concentration of 0.01% by mass or less, and the SDS is used at a concentration of 0.01% by mass or less (provided that in the method, humans are excluded from the target substrate to which the sophorolipid and the SDS are applied).

9. The method according to claim 8, wherein the biofilm is derived from Gram-negative bacteria.

10. A method for virus inactivation, characterized by using soborolipids and sodium dodecyl sulfate (SDS), wherein the virus is SARS-CoV-2, the soborolipids are used at a concentration of 0.1% by mass or less, and the SDS is used at a concentration of 0.1% by mass or less (provided that in the method, humans are excluded from the target substrate to which the soborolipids and the SDS are applied).

Images