Formulation

Abstract

The present invention relates to a composition comprising an insecticidal protein and maltodextrin having a DE of 5 to 50, wherein the composition has a pH of 5.0 to 10. The present invention also relates to a method for controlling or preventing intrusion of a plant, a part thereof, or a habitat by pests, comprising the steps of preparing a composition according to the present invention and applying the composition according to the present invention to a plant, a part thereof, or a habitat.

Description

Technology Field

[0001] The present invention relates to a composition comprising an insecticidal protein and maltodextrin, a method for preparing a composition according to the present invention, and a method for controlling or preventing intrusion of a plant, a part thereof, or a habitat by pests, comprising the step of applying a composition according to the present invention to a plant, a part thereof, or a habitat. Background Technology

[0002] Pests are primarily controlled by chemical insecticides. Biological pesticides are receiving increasing attention as an alternative to chemical pesticides.

[0003] Examples of biological insect control agents include Bacillus thuringiensis, which expresses insecticidal proteins such as delta (δ)-endotoxin (also called Cry protein) and a vegetative insect protein called Vip ( Bacillus thuringiensis These proteins are secreted into the growth medium during vegetative growth. There are three subfamilies of Vip proteins: Vip1, Vip2, and Vip3. Vip3 is the most extensively studied protein with insecticidal activity against a broad spectrum of lepidopteran pests (Syed et al, Toxins 2020, 12, 522).

[0004] WO2013 / 122720 discloses an engineered insecticidal polypeptide having activity against Ostrinia nubilalis (European corn borer).

[0005] When insecticidal proteins are used on plants, they must be formulated to remain stable for a sufficiently long period for insects to consume. Chang, L. and Pickal, MJ (2009) describe various mechanisms of protein stabilization in the solid state. It is concluded that protein stabilization in the solid state remains a controversial topic. There are no teachings or suggestions regarding methods for stabilizing proteins such as Vip. US2013 / 0296165 discloses dry stabilization compositions for live substances comprising a carbohydrate component and a protein component comprising a hydrolyzed protein. Insecticides formulated with pectin, sucrose, dibasic calcium phosphate, calcium chloride, and gluconolactone are disclosed. Additionally, a stable dry powder containing an enzyme based on a hydrogel is disclosed in US2013 / 0296165. US2013 / 0296165 does not provide teachings or suggestions regarding Vip proteins that remain stable for a sufficiently long period for insect consumption.

[0006] WO 2020182994 discloses an insecticidal composition comprising maltodextrin, which comprises at least 75% of an oligosaccharide having a chain length of 3 to 7 based on a total oligosaccharide mixture having a chain length of 3 to 20. The maltodextrin composition is active against mites.

[0007] WO 2010081815 discloses an aqueous gelatin-free, egg-free two-phase coacervate composition comprising a water-soluble biological active agent, a polysaccharide-free tension-active system, and a water-soluble carrier comprising maltodextrin, erythritol, xylitol, sorbitol, mannitol, maltitol, isomalt, and lactitol having an Mw of less than 1,800. WO 2010081815 does not provide any teaching or suggestion regarding a method for stabilizing Vip proteins.

[0008] Behle et al. (1997) disclose a Bacillus thuringiensis formulation containing wheat flour / gluten (2% by weight: volume) in the literature [J. of Economic Entomology, Vol. 90, No. 6, p. 1561-1566]. Behle et al. do not disclose or teach the composition of the insecticidal protein.

[0009] A composition containing insecticidal proteins that are stable in plants and / or sustained for a sufficiently long time for insects to consume is required.

[0010] The present invention relates to a composition comprising an insecticidal protein and maltodextrin having a DE of 5 to 50, wherein the composition has a pH of 5.0 to 10.

[0011] Surprisingly, it was found that a composition comprising insecticidal protein and maltodextrin according to the present invention can control pests for at least 3 days, preferably at least 4, 5, 6 days, or at least 7 days.

[0012] In a second aspect, the present invention relates to a method for preparing a composition according to the present invention, wherein the method

[0013] a. A step of culturing microbial cells that express insecticidal proteins;

[0014] b. A step of adding a salt solution having a pH of 5.0 to 10 to microbial cells containing an insecticidal protein;

[0015] c. Optionally, a step of lysing microbial cells; and

[0016] d. It includes the step of adding maltodextrin having a DE of 5 to 50.

[0017] In a third aspect, the present invention relates to a method for controlling or preventing intrusion of a plant, a part thereof, or a habitat by pests, comprising the step of applying a composition according to the present invention to a plant, a part thereof, or a habitat.

[0018] In a fourth aspect, the present invention relates to a plant, a part thereof, or a habitat comprising a composition according to the present invention.

[0019] In a fifth embodiment, the present invention relates to the use of maltodextrin having a DE of 5 to 50 for stabilizing insecticidal proteins for at least 3 days. Specific details for implementing the invention

[0020] The present invention relates to a composition comprising an insecticidal protein and maltodextrin having a dextrose equivalent (DE) of 5 to 50, wherein the composition has a pH of 5 to 10. Surprisingly, the composition comprising the insecticidal protein of the present invention, for at least 3 days after applying the composition to a plant, pests, e.g., Spodoptera pests, e.g., Spodoptera ritoralis ( Spodoptera littoralis It was found that it can control the Egyptian cotton beetle. Therefore, surprisingly, the composition was found to have persistence on plants for at least 3 days, preferably at least 4, 5, 6, or 7 days after application to the plants.

[0021] As used herein, a composition comprising a persistent insecticidal protein on plants is a composition comprising an insecticidal protein that is stable on plants. An insecticidal protein that persists for at least 3 days is used to indicate that the insecticidal protein exhibits activity against pests on plants for at least 3 days.

[0022] In one embodiment, the insecticidal protein is a protein that is soluble in an aqueous solution with a pH of 5 to 10, preferably 6 to 9. The aqueous solution comprises water. As used herein, a soluble protein means that the protein dissolves in an aqueous solution by forming a homogeneous mixture at the molecular level. The soluble protein comprises charged and uncharged hydrophilic residues capable of forming dipole-dipole interactions or hydrogen bond interactions with the surrounding aqueous solution.

[0023] As used herein, "insecticidal" protein or insecticidal protein means a protein that is toxic to insects or pests. The insecticidal protein may be any insecticidal protein, for example, a wild-type or naturally occurring insecticidal protein or a variant, or an engineered insecticidal protein. The insecticidal protein may be a vegetative insect protein (Vip), a Cry protein, and / or a Txp40 protein.

[0024] As used herein, the term “Cry protein” refers to a Bacillus thuringiensis crystal delta-endotoxin type insecticidal protein. The term “Cry protein” may refer to any bioactive fragment or toxin thereof, including the protoxin form, or the partially processed form and the mature toxin form, e.g., the N-terminal peptidyl fragment and / or the form without the C-terminal protoxin tail.

[0025] As used herein, "Txp40 protein" refers to two bacterial genera, Xenorabdus ( Xenorhabdus ) and Photorhabdus( Photorhabdus It is a toxin produced by ), and these are each the nematode genus Steinerenema ( Steinernema ) and heterolabuditis( Heterorhabditis It is symbiotically associated with ). Nematode-bacteria pairs can invade and kill specific insects. The Txp40 protein is photorhabdus luminescence (P. luminescensIt was first isolated from ) and was found to have injectable toxicity to various pests. The Txp40 protein is disclosed, for example, in WO2020 / 247465.

[0026] Preferably, the insecticidal protein as disclosed herein is a vegetative insecticidal protein, wherein the vegetative insecticidal protein comprises Vip1, Vip2, Vip3 and / or Vip4, preferably Vip3A and / or Vip3D. The vegetative insecticidal protein preferably comprises a Vip3A protein comprising an amino acid sequence according to SEQ ID NO. 1, or an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with the amino acid sequence according to SEQ ID NO. 1. The vegetative insecticidal protein may also comprise a Vip3D protein comprising an amino acid sequence according to SEQ ID NO. 2, or a protein comprising an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with the amino acid sequence according to SEQ ID NO. 2. The vegetative insecticidal protein may also comprise a variant of the Vip3D protein comprising the amino acid sequence according to SEQ ID NO. 3, or a protein comprising an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with the amino acid sequence according to SEQ ID NO. 3. The amino acid sequence according to SEQ ID NO. 3 corresponds to the amino acid sequence according to SEQ ID NO. 2, said amino acid sequence comprising the amino acid substitution K455A compared with SEQ ID NO. 2. The engineered vegetative insecticidal protein is disclosed, for example, in WO2013 / 12272. The amino acid sequence SEQ ID NO. 3 in WO2013 / 12272 annotated with Vip3E in WO2013 / 12272 corresponds to SEQ ID NO. 3 disclosed herein.

[0027] As used herein, the terms “percent identity” and “identical percentage” refer to the association of two or more nucleotide or amino acid sequences, which may be calculated by the steps of: (i) comparing two optimally aligned sequences across a comparison window; (ii) determining the number of positions in the two sequences where the same nucleic acid base (in the case of a nucleotide sequence) or amino acid residue (in the case of a protein) occurs, thereby calculating the number of matched positions; (iii) dividing the number of matched positions by the total number of positions in the comparison window; and then (iv) multiplying this quotient by 100% to calculate the percentage identity. Where the “percent identity” is calculated for a reference sequence without specifying a particular comparison window, the percentage identity is determined by dividing the number of matched positions across the alignment region by the total length of the reference sequence. Accordingly, for the purposes of the present invention, when two sequences (query and target) are optimally aligned (a gap is allowed in the alignment), the “percent identity” for the query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions of the query sequence (or comparison window) over the length and then multiplied by 100%.

[0028] In one embodiment, the composition according to the present invention comprises maltodextrin having 5 to 50 dextrose equivalents (DE), preferably 6 to 40 DE, preferably 7 to 35 DE, preferably 8 to 32 DE, and preferably 9 to 30 DE. Surprisingly, maltodextrin having 5 to 50 DE was found to stabilize insecticidal proteins on plants, thereby producing proteins active against insects for at least 3 days. Surprisingly, maltodextrin as disclosed herein was found to stabilize insecticidal proteins during storage and improve persistence on plant surfaces, such as on plant leaves.

[0029] The inventors have discovered that sugars such as soluble potato starch, methylcellulose, and disaccharides do not produce insecticidal proteins that are stable in plants for at least three days. Glucose as a monosaccharide has also been found to increase the persistence of insecticidal proteins in plants. However, a disadvantage of glucose as a stabilizer is that glucose is not a suitable stabilizer because it is a carbon substrate that promotes the growth of microorganisms such as bacteria or fungi.

[0030] The composition according to the present invention has a pH of 5 to 10, preferably a pH of 5.5 to 9.5, preferably a pH of 6 to 9, a pH of 6.5 to 8.8, preferably a pH of 6.8 to 8.5, preferably a pH of 7 to 8.3, and preferably a pH of 7.2 to 8. It has been found that the pH of the composition affects the activity of the insecticidal protein. A vegetative insecticidal protein, such as Vip3A or Vip3D as disclosed herein, has been found to be active at a pH of 6 to 10, preferably a pH of 6.5 to 9.5, preferably a pH of 7 to 9, preferably a pH of 7.2 to 8.8, and preferably a pH of 7.5 to 8.5. The vegetative insecticidal protein is soluble at a pH of 5 to 10.

[0031] In one embodiment, the composition according to the present invention comprises an insecticidal protein and a maltodextrin having a DE of 5 to 50 in a weight / weight ratio of insecticidal protein to maltodextrin having a DE of 5 to 50, preferably 1:200 to 1:1, preferably 1:150 to 1:2, preferably 1:100 to 1:5, preferably 1:80 to 1:8, preferably 1:70 to 1:10, preferably 1:60 to 1:15, preferably 1:50 to 1:20, preferably 1:40 to 1:25.

[0032] In a further embodiment, the composition disclosed herein comprises an agro-acceptable carrier. As used herein, an "agro-acceptable carrier" may include natural or synthetic, organic or inorganic materials that facilitate application to plants, or parts thereof, or habitats in combination with an active protein. Examples of agro-acceptable carriers include, without limitation, powders, dusts, pellets, granules, sprays, emulsions, colloids, and solutions. Agro-acceptable carriers may further include, but are not limited to, inactive components, dispersants, surfactants, adjuvants, tackifiers, adhesives, binders, or combinations thereof that may be used in agricultural formulations. Suitable surfactants may include, for example, non-ionic surfactants, such as butyl-block polyalkylene oxide copolymer Toximul®8320 or polysorbate 20, such as Tween®20, or alcohol ethoxylates, such as Synperonic®10 / 6. This composition may be applied in any manner that brings insecticidal proteins or other pest control agents into contact with pests. Accordingly, the composition may be applied to the surface of a plant or a part of a plant, including seeds, leaves, flowers, stems, tubers, roots, etc.

[0033] In one embodiment, the composition according to the present invention comprises a salt. Any suitable salt, preferably a buffer salt, may be used. Preferably, the buffer salt produces a composition having a pH of 5 to 10. Preferably, the salt comprises a phosphate salt, for example, a phosphate salt and sodium chloride, ammonium bicarbonate, or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), or tris(hydroxymethyl)aminomethane (TRIS).

[0034] The compositions disclosed herein may be used as they are, as tank mixes, or conveniently formulated in known ways as emulsifiable concentrates, coatable pastes, directly sprayable or dilutable solutions or suspensions, diluted emulsions, wettable powders, soluble powders, dusts, granules, and also, for example, in the form of encapsulations within polymeric materials. As with the types of compositions, the method of application, such as spraying, atomizing, dusting, spreading, coating, or injection, is selected according to the intended purpose and general circumstances. The compositions may also contain additional adjuvants, such as stabilizers, antifoaming agents, viscosity modifiers, binders, or tackifiers, as well as fertilizers, micronutrient feeds, or other formulations to achieve special effects.

[0035] In one embodiment, the composition according to the present invention may be a soluble powder or tank mixture known to those skilled in the art. A soluble powder is an insecticide formulation in which an active ingredient, such as the insecticidal protein disclosed herein, exists in powder form (very similar to a wettable powder) and can be mixed into a complete solution. Soluble powder formulations are also known as water-soluble powders or water-soluble packets (WSP). The tank mix used herein is a composition obtained by mixing all components constituting the tank mix into a single tank to perform a single spray application. The tank mix may also be referred to as a soluble concentrate.

[0036] In a further embodiment, the insecticidal composition comprises microbial cells, preferably, the microbial cells produce an insecticidal protein in the composition disclosed herein. Preferably, the microbial cells are inactivated or lysed. Preferably, the microbial cells are non-living, inactivated, or lysed microbial cells. Such insecticidal compositions may be prepared by drying, freeze-drying, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture comprising microorganisms expressing the insecticidal protein disclosed herein. The composition disclosed herein may comprise at least about 1 wt%, preferably at least 5, at least 10, at least 20, at least 25, at least 30, at least 35, at least 40, at least 50, at least about 60, at least 70, at least 80, at least 90, at least 95, at least 97, or at least 99 wt% based on the protein of the present invention. In a further embodiment, the composition comprises about 1 to about 99 wt% of the insecticidal protein of the present invention.

[0037] In one embodiment, the present invention relates to a method for preparing a composition according to the present invention, wherein the method

[0038] a. A step of culturing microbial cells expressing insecticidal proteins;

[0039] b. A step of adding a salt solution having a pH of 5.0 to 10 to microbial cells containing an insecticidal protein;

[0040] c. A step of selectively lysing microbial cells; and

[0041] d. Includes the step of adding maltodextrin having a DE of 5 to 50.

[0042] The terms "microbial cell" or "microorganism" are used interchangeably herein. Microbial cells or microorganisms expressing insecticidal proteins may be naturally occurring microorganisms or genetically modified or recombinant microorganisms. Microorganisms are Bacillus species ( Bacillus sp.), for example, Bacillus thuringiensis ( Bacillus thuringiensis ), Esherikia Kolai( Escherichia coli ), or yeast, for example, Piquia Pastoris ( Pichia pastoris ) or Aspergillus niger( Aspergillus niger ) or Aspergillus oryzae( Aspergillus oryzae It may be a filamentous fungus such as Aspergillus species. Preferably, the microorganism is a microorganism expressing a gene encoding the Vip protein, and preferably, the microorganism is a genetically modified E. coli with a nucleic acid sequence encoding the Vip protein. E. coli) and, where the Vip protein is Vip1, Vip2, Vip3 and / or Vip4, preferably Vip3A and / or Vip3D. The Vip protein may be a naturally occurring or wild-type protein or a variant or an engineered protein. Preferably, the microorganism expresses at least one Vip3A protein comprising an amino acid sequence according to SEQ ID NO. 1, or an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with the amino acid sequence according to SEQ ID NO. 1. The microorganism may also express a Vip3D protein comprising an amino acid sequence according to SEQ ID NO. 2, or a protein comprising an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with the amino acid sequence according to SEQ ID NO. 2. The microorganism may also express a variant of the Vip3D protein comprising the amino acid sequence according to SEQ ID NO. 3, or a protein comprising an amino acid sequence having at least 80%, preferably at least 85%, 90%, 95%, preferably at least 96%, 97%, 98%, or at least 99% identity with respect to the amino acid sequence according to SEQ ID NO. 3. The amino acid sequence according to SEQ ID NO. 3 corresponds to the amino acid sequence according to SEQ ID NO. 2, wherein the amino acid sequence comprises the amino acid substitution K455A compared to SEQ ID NO. 2.

[0043] As used herein, the term “recombinant” generally refers to a nucleic acid (e.g., DNA or RNA) or protein or organismal form that is not found in nature and is therefore produced by human intervention. As used herein, “recombinant microorganism” is a microorganism that does not exist in nature, is the result of human intervention, and contains an introduced gene or a heterologous nucleic acid molecule integrated into its genome.

[0044] "Transformed / transformed / recombined" refers to a host organism, such as a microorganism, into which a heterologous nucleic acid molecule has been introduced. The nucleic acid molecule may be stably integrated into the host genome, or it may also exist as an extrachromosomal molecule. "Non-transformed," "non-transformed," or "non-recombined" hosts refer to wild-type organisms, such as microorganisms, that do not contain the heterologous nucleic acid molecule.

[0045] The culture of microbial cells may be carried out by methods known in the art. Typically, the culture of microbial cells is carried out by batch culture, fed-batch culture, or continuous culture. The culture of microbial cells comprises the step of culturing microbial cells in a fermentation medium containing suitable carbon and nitrogen sources. Suitable carbon sources may include molasses, e.g., sugar beet or sugarcane molasses, polysaccharides, wheat flour, starch, sugar, or glucose. Suitable nitrogen sources may be casein hydrolysate, tryptone, ammonium sulfate, ammonia, yeast extract, peptone, or urea. The culture of microorganisms may be carried out under aerobic and / or anaerobic conditions.

[0046] The cultivation of microbial cells expressing an insecticidal protein typically includes the step of producing a fermented broth containing microbial cells containing the insecticidal protein.

[0047] Preferably, the method according to the present invention comprises the step of obtaining microbial cells containing an insecticidal protein. The obtaining of microbial cells, preferably microbial cells containing an insecticidal protein, may be carried out by methods known in the art, such as centrifugation or filtration. The insecticidal protein may be an intracellular and / or extracellular protein of the microbial cells. In one embodiment, the method according to the present invention comprises the step of obtaining microbial cells, preferably microbial cells and an insecticidal protein, from a fermentation broth before adding a salt solution to the microbial cells.

[0048] The method according to the present invention further comprises the step of adding a salt solution having a pH of 5.0 to 10 to microbial cells containing an insecticidal protein. Preferably, the salt solution is a salt buffer solution having a pH of 5.5 to 9.5, preferably 6 to 9, a pH of 6.5 to 8.5, and preferably 7 to 8. Preferably, the salt solution is a solution of a phosphate salt, for example, a phosphate salt and sodium chloride, ammonium carbonate, or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS). Preferably, the salt solution is added at a concentration of 1 to 100 mM, for example 5 to 80 mM, for example 10 to 60 mM.

[0049] The method according to the present invention preferably comprises a step of lysing microbial cells. The lysing or lysis of microbial cells may be performed by any suitable known technique, for example, microfluidics using a French press, mechanical lysing using a hydraulic pump, or sonication using a ball mill. The step of lysing microbial cells may further comprise a step of removing the lysed cells, such as cell debris or cellular material, by, for example, centrifugation or filtration, to obtain a purified solution containing insecticidal proteins.

[0050] The method according to the present invention further comprises the step of adding maltodextrin having a DE of 5 to 50. Preferably, the maltodextrin has a DE of 6 to 40, preferably a DE of 7 to 35, preferably a DE of 8 to 32, and preferably a DE of 9 to 30. The maltodextrin added to the method disclosed herein may be an aqueous solution containing maltodextrin or maltodextrin powder.

[0051] In one embodiment, the method according to the present invention further comprises the step of removing water after steps a), b), c) and / or d).

[0052] Water may be removed during or after steps a), b), c), and / or d) using techniques known in the art. Known techniques for removing water are, for example, freeze-drying, spray drying, spray coagulation, or spray freeze-drying.

[0053] The method according to the present invention may further include the step of adding an agriculturally acceptable carrier. Any suitable agriculturally acceptable carrier may be used and may be a carrier defined herein.

[0054] The composition according to the present invention can be obtained by the method according to the present invention.

[0055] In a third aspect, the present invention relates to a method for controlling or preventing intrusion of a plant, a part thereof, or a habitat by pests, comprising the step of applying a composition according to the present invention to a plant, a part thereof, or a habitat.

[0056] In addition, a pest control method comprising the step of bringing a pest into contact with a composition according to the present invention is disclosed herein.

[0057] The method according to the present invention does not include a method of treating a human or animal body by surgery or a treatment method, or a diagnostic method performed on a human or animal body.

[0058] The method for controlling or preventing pest infestation disclosed herein comprises the step of controlling or preventing pests on a plant, a part thereof, or a habitat, and the method comprises the step of applying a composition according to the present invention to the plant, a part thereof, or a habitat.

[0059] The step of applying the composition according to the present invention to a plant, a part thereof, or a habitat preferably includes the step of bringing pests into contact with the composition according to the present invention.

[0060] The step of bringing a pest into contact with the composition disclosed herein comprises delivering the composition to the insect. “Delivering” a composition containing an insecticidal protein means that the insecticidal protein comes into contact with the insect to cause a toxic effect and control of the insect. The pests used herein include insect larvae and adult insects, and preferably the pests include larvae.

[0061] A composition containing an insecticidal protein provided control of pests or prevented infestation for at least 3 days, preferably at least 4, 5, 6 days, or at least 7 days. Surprisingly, the composition containing the insecticidal protein of the present invention was found to have persistence on plants, parts thereof, or habitats for at least 3 days, preferably at least 4, 5, 6 days, or at least 7 days after the composition was applied to plants, parts thereof, or habitats.

[0062] A composition containing insecticidal protein is applied in a suitable amount. The suitable amount is 2 to 200 g of protein per hectare (ha), e.g. 5 to 150 g of protein per ha, or 10 to 100 g per ha, or 15 to 75 g of protein per ha.

[0063] The step of applying the insecticidal protein to a plant, a part thereof, or habitat may be carried out by any suitable method in the art. Suitable methods of application to the plant surface, or of contact with the plant, a part thereof, or habitat, include spraying, atomization, acid dispersion, scattering, misting, spraying, coating, immersion, or injection, depending on the intended purpose and general circumstances. The composition may also contain additional adjuvants, such as stabilizers, antifoaming agents, viscosity modifiers, binders, or tackifiers, as well as fertilizers, micronutrient sources, or other formulations to achieve special effects.

[0064] In one embodiment, the method according to the present invention comprises the step of controlling pests, wherein the pests are Coleopteran, Hemipteran, or Lepidopteran pests. Preferably, the pests are Lepidopteran pests, such as Austrian nubilalis ( Ostrinia nubilalis )(European corn borer), Flutella xylostella( Plutella xylostella )(Cabbage moth), Spodoroptera prugiferda( Spodoptera frugiperda )(fall armyworm), spodoptera litoralis( Spodoptera littoralis )(Egyptian cotton beetle), Spodoptera eridania( Spodoptera eridania )(Southern cutworm), Spodoptera exigua( Spodoptera exigua )(Eastern armyworm), Spodoptera ritura( Spodoptera litura )(Tobacco cutworm), Agrotis epsilon( Agrotis ipsilon )(Agrotis orthogonia), Agrotis orthogonia( Agrotis orthogonia )(Pale Western Cutworm), Striacosta albicosta( Striacosta albicosta )(Western Bean Cutworm), Helicobacter pupae Zea( Helicoverpa zea )(Giant Tobacco Moth), Helicobacter armigera( Helicoverpa armigera )(cotton ball moth), Heliotis virecens( Heliothis virescens )(tobacco moth), Helicobacter phallus punctigera( Helicoverpa punctigera )(Australian tobacco moth), Manduka sexta( Manduca sexta )(Tobacco hawk moth), Trichophucia ni( Trichoplusia ni )(Cabbage moth), Mamestra brasicae( Mamestra brassicae )(Cabbage moth), Tuta Absolute( Tuta absoluta )(Tomato hornworm), Lobesia botrana(European grape leaf roller), Grapolita molesta( Grapholita molesta )(Oriental fruit moth), Eupoecilia ambiguella(vine vine moth), Cydia pomonella( Cydia pomonella )(Apple leafminer), Chrysodeix includens( Chrysodeixis includens )(Soybean Silver Moth), Chilo Surfresalis( Chilo suppressalis)(Striped rice stem moth), Pectinophora gossyphiella( Pectinophora gossypiella )(Pinkball Moth), Diatrae grandiocella( Diatraea grandiosella )(Southwestern corn borer), Diatrae saccharalis( Diatraea saccharalis )(Sugarcane moth), Elasmopulpus lignocellus( Elasmopalpus lignosellus )(small corn cob moth), Anticarcia gemmatalis( Anticarsia gemmatalis )(Velvet Bean Cutworm), Platypheña Scarab( Plathypena scabra They are )(green clover beetle), and Cochylis hospes (striped sunflower moth).

[0065] Preferably, the pests are Austriania nubilalis (European corn borer), Flutella xylostella (diamondworm), Spodoroptera prugiferda (fall armyworm), Spodoptera litoralis (Egyptian cotton beetle), Spodoptera eridania (Southern cutworm), Spodoptera exigua (Ox armyworm), Spodoptera ritura (Tobacco cutworm), Agrotis epsilon (Black cutworm), Helicobacter zea (Giant tobacco moth), Helicobacter zea (Cotton ball moth), Heliotis viressens (Tobacco moth larva), Trichoplushia ni (Cabbage silver-striped moth), Mamestra brasicae (Cabbage moth), Tuta absolute (Tomato hornworm), Robesia botrana (European grape leafroller), Grapolita molesta (Oriental fruit moth), Euphorecilia ambiguela (Vine vine moth), Cydia pomonella (Apple leafminer), Chrysodeixis They are *Includens* (soybean silver moth) and *Chilo supresalis* (striped rice stem moth).

[0066] Preferably, the pest is a Spodoptera pest, preferably Spodoptera ritoralis (Egyptian cotton beetle).

[0067] The term "plant" refers to all physical parts of a plant, including seeds, seedlings, saplings, roots, tubers, stems, stalks, leaves, and fruits.

[0068] The term "plant" includes "useful plant" or "crop." The expressions "useful plant" and "crop" are used interchangeably herein.

[0069] Plants include corn, soybeans, rice, wheat, barley, rye, oats, sorghum, millet, sunflowers, safflower, sugar beets, cotton, sugarcane, rapeseed, alfalfa, tobacco, peanuts, vegetables (including sweet potatoes, beans, peas, chicory, lettuce, cabbage, cauliflower, broccoli, turnips, carrots, eggplants, cucumbers, radishes, spinach, potatoes, tomatoes, asparagus, onions, garlic, melons, peppers, celery, squash varieties, pumpkins, zucchini, etc.), fruits (including apples, pears, quinces, plums, cherries, peaches, nectarines, apricots, strawberries, grapes, raspberries, blackberries, pineapples, avocados, papayas, mangoes, bananas, etc.), and special plants (e.g., Arabidopsis thaliana ( Arabidopsis )), or woody plants (e.g., conifers and / or deciduous trees) are included, but are not limited thereto. In an embodiment, the plant in the method of the present invention is a crop such as corn, sorghum, wheat, sunflower, tomato, cruciferous plant, pepper, potato, cotton, rice, soybean, sugar beet, sugarcane, tobacco, barley, rapeseed plant, etc. Preferably, the plant includes soybean and / or cotton.

[0070] The term "useful plant" should also be understood to include useful plants that have become resistant to herbicides or classes of herbicides such as bromoxinyl (e.g., HPPD inhibitors, ALS inhibitors, e.g., primisulfuron, prosulfuron and triploxulfuron, EPSPS (5-enol-pyrovyl-schikimate-3-phosphate-synthase) inhibitors, GS (glutamine synthase) inhibitors or PPO (protoporpyrinogen-oxidase) inhibitors) as a result of traditional breeding methods or genetic engineering.

[0071] The term “part” of a plant includes seeds, seedlings, saplings, roots, tubers, stems, stalks, leaves, and fruits.

[0072] As used in this specification, the term “habitat” means a field in which plants are growing or on which they are growing, a place where seeds of cultivated plants are sown, or a place where seeds are placed in the soil.

[0073] In a fourth aspect, the present invention relates to a plant, part thereof, or habitat comprising a composition according to the present invention. Surprisingly, it was found that a plant, part thereof, or habitat comprising a composition according to the present invention suffered less or no damage from pests for at least 3 days, preferably at least 4, 5, 6, or 7 days, compared to a plant, part thereof, or habitat not comprising a composition according to the present invention.

[0074] In one additional aspect, the present invention relates to the use of maltodextrin having a DE of 5 to 50 for stabilizing insecticidal proteins for at least 3 days, preferably at least 4, 5, 6 days, or at least 7 days. Preferably, the use comprises stabilizing insect proteins in plants, parts thereof, or habitats. The use further comprises a salt solution having a pH of 5.0 to 10. All embodiments described above are applicable to uses according to the present invention.

[0075] Examples

[0076] method

[0077] Fermentation production of the variant Vip3D (Sequence No. 3) having a substitution at position K455A compared with Vip3A (Sequence No. 1) and Sequence No. 2.

[0078] To produce Vip3A (Sequence No. 1) and Vip3D variant proteins (Sequence No. 3), the literature [Estruch et al 1 and Khan et. al 2 An Escherichia coli strain capable of expressing the Vip3 protein was generated following the method disclosed in ]. The nucleic acid sequence for the Vip3 protein is this. coli ( E. coliCodons were optimized for expression in ), and cloned into a standard protein expression vector. A vector containing the Vip3 encoding sequence was introduced into an E. coli protein-expressing host via chemical transformation to provide a recombinant strain for Vip3 protein production. Using a stirred tank bioreactor, [Estruch et al. 1 and Khan et. al 2 Recombinant E. coli strains were cultured by batch fermentation in a complex liquid medium initiated in ]. Vip3 protein expression was induced by the addition of IPTG, and after continuing incubation for an additional period, biomass was obtained by centrifugation.

[0079] References:

[0080] 1) Estruch JJ, Warren GW, Mullins MA, Nye GJ, Craig JA, Koziel MG. Vip3A, a novel Bacillus thuringiensis vegetative insecticidal protein with a wide spectrum of activities against lepidopteran insects. Proc Natl Acad Sci US A. 1996 May 28;93(11):5389-94.

[0081] 2) MH Khan, G. Jander, Z. Mukhtar, M. Arshad, M. Sarwar, S. Asad, Comparison of in Vitro and in Planta Toxicity of Vip3A for Lepidopteran Herbivores, Journal of Economic Entomology, Volume 113, Issue 6, December 2020, Pages 2959-2971.

[0082] Recovery of Vip3 protein

[0083] Cell mass was collected from the fermentation broth using a disk stack centrifuge. Unless otherwise specified, the cell mass was resuspended in various intensities of aqueous sodium phosphate or Tris buffer and 137 mM sodium chloride or Tris buffer. Cells were lysed using a microfluidic device. Cell debris was removed by centrifugation followed by chemical inactivation of the cells using a biocide (benzisothiazolinone, BIT).

[0084] The purified material contained soluble protein at a concentration of 2 to 10 mg / ml, as measured by SDS-PAGE and densitometric method against purified standard material.

[0085] Preparation of Vip3 / Sugar Soluble Concentrate (SL) or Tank Mix

[0086] Purified Vip3 soluble solution as a solution or freeze-dried powder was directly added to a stock solution at a weight ratio of Vip3 to per 10, 20, or 30 wt / wt% (see Table 1 below) at a ratio of 1:2, 1:4, 1:10, 1:20, 1:30, 1:50, or 1:100.

[0087] Preparation of spray tank: A solution of the above-mentioned soluble concentrate (SL) was diluted with water to achieve an accurate application rate (g AI / ha) and constitute 90% of the spray volume, and finally, an adjuvant (maintenance adjuvant, wetting agent, rain resistance enhancer, photoprotector) was added as a tank mix to achieve an appropriate concentration for the remaining 10% of the spray volume.

[0088] Sugars used to stabilize Vip protein in plants sugar suppliers Maltodextrin (Glucidex 9) Roquette Maltodextrin (Glucidex 12) Rocket Maltodextrin (Glucidex 17) Rocket Maltodextrin (Glucidex 21) Rocket Maltodextrin (Glucidex 29) Rocket Lactose DFE Pharma International Maltos Sigma Aldrich Trehalos Sigma Aldrich Sucrose Sigma Aldrich glucose Sigma Aldrich potato starch Sigma Aldrich Methocell A 15 LV (methylcellulose) Merck Life Science

[0089] 1- Preparation of Vip3 / Sugar-Soluble Powder (SP) Formulation

[0090] Sugar (see Table 1) was directly dissolved in a Vip3 purification solution at a weight ratio of Vip3 to sugar of 1:10, 1:30, 1:50, or 1:100, and water was removed through freeze-drying to prepare a soluble powder formulation.

[0091] Preparation of spray tank: The above-mentioned soluble powder (SP) was added and dissolved in water to achieve an accurate application rate (g AI / ha) and constitute 90% of the spray volume, and finally, an adjuvant (e.g., a maintenance adjuvant, a wetting agent, a rain resistance enhancer, or a photoprotector) was added as a tank mix if necessary to achieve an appropriate concentration of the remaining 10% of the spray volume.

[0092] Continuous Activity Analysis:

[0093] Soybeans (Glycine Max) Glycine max )) or cotton plant (Gossypium hysutum ( Gossypium hirsutum The diluted test solution was sprayed in the application chamber. Plants were incubated in a greenhouse under controlled conditions (temperature of 22°C during the day and 20°C at night, and relative humidity of 65%).

[0094] Then, the leaves were cut and placed in Petri dishes with wet filter paper in various cases.

[0095] Petri dishes were infested with the Egyptian cotton beetle *Spodoptera ritoralis* (8-10 L2 larvae per dish) a few hours after infestation or 1, 3, 4, or 5 days after the day of infestation (DAA), and covered with a fiber filter and a plastic lid. The larvae were evaluated for mortality in each infestation case 5 days after infestation.

[0096] All analyses were performed in the laboratory under controlled environmental conditions (temperature 25°C and relative humidity 65%, 16 hours of light and 8 hours of darkness).

[0097] The larval mortality rate for the corresponding control response in the activity persistence analysis was corrected using Abbott's formula (test mortality % - control mortality % / 100 - control mortality x 100) (WS Abbott, A Method of Computing the Effectiveness of an Insecticide, Journal of Economic Entomology, Volume 18, Issue 2, 1 April 1925, Pages 265-267).

[0098] Example 1. Effects of Maltodextrin and Disaccharides on Vip3A Stability

[0099] A solution of the purified Vip3 lysate was added to a 10% maltodextrin or disaccharide solution to prepare a soluble concentrate (SL) type formulation. A 1x PBS buffer solution (10 mM sodium phosphate and 137 mM sodium chloride) at pH 7.4 was used during cell resuspension. Soybean plants were treated with a 75 ppm solution of Vip3A and maltodextrin (Table 2) and a 37.5 ppm solution of Vip3A and different disaccharides (Table 3) as described above.

[0100] Cotton plants were treated with 200 ppm of Vip3A and polysaccharides (maltodextrin) and disaccharides (maltose and lactose) (Table 4). Karate (lambda cyhalothrin) was used as a positive control.

[0101] The results in Table 2 show that the presence of maltodextrin with DEs of 5 to 50 protected the Vip3A protein and provided control over Spodoptera ritoralis L2 larvae for up to 3 days after application to soybean plants. The dextrose equivalent values ​​of the maltodextrin types used in the experiment had a negligible effect on the persistence of the Vip3A protein on soybean leaves (Table 2).

[0102] The results in Table 3 indicate that the presence of disaccharides protected the Vip3A protein and provided control of Spodoptera ritoralis L2 larvae on soybean leaves for only 1 day. Control against Spodoptera ritoralis was very weak when larvae invaded the leaves 3 days after the application of the composition of disaccharides and Vip3A.

[0103] The results in Table 4 show that the addition of maltodextrin to Vip3A resulted in longer persistence of Vip3A on cotton leaves (up to 4 DAA) compared to maltose and lactose.

[0104] The mortality rate of Spodoptera ritoralis L2 larvae exposed to 0, 1, and 3 DAAs was evaluated 5 days after infesting soybean leaves treated with Vip3A along with maltodextrin. test compound ratio Sugar type AI weight ratio invasion 0 DAA 1 DAA 3 DAA Control rate % Control rate % Control rate % Vip3A 　 75 ppm (37.5 g AI / ha) - - 100 65 0 Glusidex 29 1:10 100 100 92 1:2 100 93 68 Glusidex 21 1:10 100 100 100 1:2 100 100 58 Glusidex 12 1:10 100 100 90 1:2 100 85 46 Glusidex 9 1:10 100 100 90 1:2 100 98 26 Karate (Lambda Cyhalothrin) 50 ppm (25 g AI / ha) - 100 100 100 Glusidex 29 0.1% - - 0 0 0 Glusidex 21 0.1% - - 5 2.5 0 Glusidex 12 0.1% - - 0 0 0 Glusidex 9 0.1% - - 3 0 0 Inspection (water) - - - - - Inspection (Water) - Not Modified - - 0 0 10

[0105] The mortality rate of Spodoptera ritoralis L2 larvae exposed to 0, 1, and 3 DAAs was evaluated 5 days after infestation of soybean leaves treated with Vip3A along with 4 disaccharides. test compound ratio Sugar type AI weight ratio invasion 0 DAA 1 DAA 3 DAA Control rate % Control rate % Control rate % Vip3A 37.5 ppm (18.75 g AI / ha) - - 70 6 0 Maltos 1:20 100 94 23 1:4 95 59 3 Sucrose 1:20 100 100 18 1:4 100 53 3 Lactose 1:20 100 75 20 1:4 93 75 5 Trehalos 1:20 100 80 35 1:4 98 53 0 Karate (Lambda Cyhalothrin) 50(25 g AI / ha) - 100 100 100 Maltose 0.1% - - 0 0 0 Sucrose 0.1% - - 0 0 0 Lactose 0.1% - - 0 0 0 Trehalose D 0.1% - - 0 0 0 Inspection (water) - - - - - Inspection (Water) - Not Modified - - 0 0 0

[0106] The mortality rate of Spodoptera ritoralis L2 larvae infested with 0, 3, and 4 DAAs was evaluated 5 days after infestation on cotton leaves treated with Vip3A with maltodextrin and disaccharides added in a ratio of 2. test compound ratio Sugar type AI weight ratio invasion 0 DAA 3 DAA 4 DAA ppm g AI / ha Control rate % Control rate % Control rate % Vip3A 200 100 - 100 0 3 200 100 Glusidex 29 1:50 100 78 83 1:5 100 43 18 200 100 Glusidex 9 1:50 100 95 73 1:5 100 45 20 200 100 Maltos 1:50 100 75 50 1:5 100 13 8 200 100 Lactose 1:50 100 60 53 1:5 100 28 5 Karate (Lambda Cyhalothrin) 50 25 - 100 100 100 Glusidex 29 1% - - 0 0 0 Glusidex 9 1% - - 0 0 10 Maltose 1% - - 0 0 0 Lactose 1% - - 0 0 3 Inspection (water) - - - - - Inspection (Water) - Not Modified - - - 2.5 0 0

[0107] Example 2. Effect of Maltodextrin on the Stability of Vip3A and Variant Vip3D

[0108] A soluble concentrate (SL) type formulation prepared by adding a purified Vip3 lysate solution to a 10% maltodextrin solution was used in this example. A 1x PBS buffer solution (10 mM sodium phosphate and 137 mM sodium chloride) at pH 7.4 was used during cell resuspension. Soybean plants were treated with Vip3A or Vip3D solutions containing 75 ppm of maltodextrin glusidex 29 as described above (Table 5). Karate (lambda cyhalothrin) was used as a positive control.

[0109] The results in Table 5 indicate that Glusidex 29, with a Vip3-to-sugar weight ratio of 1:100, stabilized both Vip3A and Vip3D variants of the plant and provided control against Spodoptera ritoralis larvae for up to 7 days after application to soybean plants.

[0110] The mortality rate of Spodoptera ritoralis L2 larvae exposed to 0, 3, 5, and 7 DAAs was evaluated after 5 days of infestation into soybean leaves treated with Vip3A- and Vip3D variant K455A (SEQ No. 3) with or without the addition of Glusidex 29. test compound ratio Sugar type AI weight ratio invasion 0 DAA 3 DAA 5 DAA 7 DAA Control rate % Control rate % Control rate % Control rate % Vip3A 75 ppm (37.5 g AI / ha) doesn't exist 100 85 25 6 Glusidex 29 1　:100 100 100 98 100 Vip3D variant Sequence No. 3 doesn't exist 100 7.5 0 0 Glusidex 29 1　:100 100 100 88 91 Karate (Lambda Cyhalothrin) 50 ppm (25 g AI / ha) - 100 100 100 100 Inspection (water) - 0 0 0 0

[0111] Example 3. Effect of different sugars on the stability of Vip3A

[0112] A soluble concentrate (SL) type formulation prepared by adding a purified Vip3 lysate solution to a 10% sugar solution (potato starch, methylcellulose, glucose, and Glusidex 29) was used in this example. A 1xPBS buffer solution (10 mM sodium phosphate and 137 mM sodium chloride) at pH 7.4 was used during cell resuspension.

[0113] As described above, soybean plants were treated with Vip3A solution in the presence of various types of sugars (see Tables 1 and 6) at 37.5 and 9.375 ppm (Table 6). Karate (lambda cyhalothrin) was used as a positive control.

[0114] The results in Table 6 indicate that among all sugars tested, maltodextrin glusidex 29 D stabilized Vip3 when applied at 37.5 ppm and 9.375 ppm, providing sustained control against Spodoptera ritoralis for up to 5 days when applied to soybean plants. Potato starch and methylcellulose did not provide excellent sustained control against Spodoptera ritoralis larvae at concentrations of 37.5 and 9.375 ppm, and glucose was less effective against Spodoptera ritoralis larvae at 9.375 ppm compared to maltodextrin glusidex 29.

[0115] Potato starch, methylcellulose, and glucose applied independently to soybean plants at 0.375% did not show activity against Spodoptera ritoralis larvae.

[0116] The mortality rate of Spodoptera ritoralis L2 larvae exposed to 0, 3, and 5 DAAs was evaluated 5 days after infesting Vip3A-treated soybean leaves with or without the addition of Glusidex 29 and other sugars. test compound ratio Sugar type invasion 0 DAA 3 DAA 5 DAA Control rate % Control rate % Control rate % ppm g AI / ha Vip3A 37.5 18.75 - 100 5 0 9.375 4.69 33 0 0 37.5 18.75 Glusidex 29 100 83 100 9.375 4.69 83 75 58 37.5 18.75 potato starch 93 25 3 9.375 4.69 13 0 0 37.5 18.75 Methylcellulose 100 40 18 9.375 4.69 80 0 5 37.5 18.75 glucose 98 98 100 9.375 4.69 98 75 8 Karate (Lambda Cyhalothrin) 50 25 100 100 100 soluble starch 0.375% - 0 0 3 Methylcellulose 0.375% - 18 0 0 glucose 0.375% - 0 0 3 Inspection (water) - - - - - Inspection (Water) - Not Modified - - 0 0 0

[0117] All sugars were added in a weight ratio of 1:100 (AI:sugar).

[0118] Example 4. Effect of salt / buffer solution on protein activity

[0119] Different buffer types and protein (AI) to protein ratios were tested in this example. As previously mentioned, biomass collected from the fermenter was resuspended in 50 mM sodium phosphate or 50 mM Tris-buffered solution and 137 mM sodium chloride at pH 8. Cells were lysed, and cell debris was removed by centrifugation.

[0120] A soluble powder type formulation was prepared by mixing the purified Vip3 solution in a 20% Glusidex 29 solution and then freeze-drying to remove water.

[0121] Soybean plants were treated by dissolving the above powder at a dose of 75 ppm Vip3A (Tables 7 and 8). Karate (lambda cyhalothrin) was used as a positive control.

[0122] The results in Tables 7 and 8 show that the sodium phosphate solution increased the stability of Vip3A compared to the Tris-buffer. By incorporating Glusidex 29 with Vip3A into the sodium phosphate buffer, control of Spodoptera ritoralis larvae was provided up to 5 DAA (Table 7).

[0123] Incorporation of Glusidex 29 with Vip3A into Tris buffer provided control of up to 3 DAA against Spodoptera ritoralis larvae. Protein activity was significantly reduced when larvae were infected with 5 DAA on the leaves (Table 8).

[0124] The mortality rate of Spodoptera ritoralis L2 larvae infected with 0, 3, and 5 DAAs was evaluated 5 days after invasion into soybean leaves treated with Vip3A mixed with Glusidex 29 at different ratios in sodium phosphate buffer solution. test compound buffer Weight ratio of incorporated Glusidex 29 (AI:sugar) invasion 0 DAA 3 DAA 5 DAA Control rate % Control rate % Control rate % Vip3A75 ppm(37.5 g AI / ha) 50 mM Tris pH 8 1:30 100 100 63 1:50 100 100 90 Karate (lambda cyhalothrin) 50 ppm (25 g AI / ha) - 100 100 100 Inspection (water) - 0 0 0

[0125] The mortality rate of Spodoptera ritoralis L2 larvae infected with 0, 3, and 5 DAAs was evaluated 5 days after infesting soybean leaves treated with Vip3A along with Glusidex 29 D mixed in different proportions in Tris buffer. test compound buffer Weight ratio of incorporated Glusidex 29 (AI:sugar) invasion 0 DAA 3 DAA 5 DAA Control rate % Control rate % Control rate % Vip3A75 ppm(37.5 g AI / ha) 50 mM sodium phosphate pH 8 1:30 100 100 63 1:50 100 100 90 Karate (lambda cyhalothrin) 50 ppm (25 g AI / ha) - 100 100 100 Inspection (water) - 0 0 0

Claims

Claim 1 A composition comprising an insecticidal protein and maltodextrin having a DE of 5 to 50, wherein the pH is 5.0 to 10. Claim 2 A composition according to claim 1, wherein the insecticidal protein is soluble in an aqueous solution with a pH of 5 to 10. Claim 3 A composition according to claim 1 or 2, wherein the insecticidal protein comprises a vegetative insecticidal protein, and the vegetative insecticidal protein comprises Vip1, Vip2, Vip3 and / or Vip4, preferably Vip3A and / or Vip3D. Claim 4 A composition according to any one of claims 1 to 3, wherein the insecticidal protein and maltodextrin have an insecticidal protein to maltodextrin ratio of 1:200 to 1:1 by weight:weight. Claim 5 A composition comprising, in any one of paragraphs 1 to 4, additionally a salt. Claim 6 A composition comprising, in any one of paragraphs 1 to 5, an agriculturally acceptable carrier. Claim 7 A composition that is a soluble powder or tank mix in any one of claims 1 to 6. Claim 8 A method for manufacturing a composition according to any one of claims 1 to 7, a. A step of culturing microbial cells that express insecticidal proteins; b. A step of adding a salt solution having a pH of 5.0 to 10 to microbial cells containing an insecticidal protein; c. A step of selectively lysing microbial cells; and d. A method comprising the step of adding maltodextrin having a DE of 5 to 50. Claim 9 A method according to claim 8, further comprising the step of removing water after steps a), b), c) and / or d). Claim 10 A composition or method according to any one of claims 1 to 7, or a method according to any one of claim 8 or 9, wherein the salt comprises sodium phosphate salt, ammonium bicarbonate, N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), or tris(hydroxymethyl)aminomethane (Tris). Claim 11 A method for controlling or preventing intrusion of a plant, part thereof, or habitat by pests, comprising the step of applying a composition according to any one of claims 1 to 7 to a plant, part thereof, or habitat. Claim 12 In claim 11, the pest is Spodoptera, preferably Spodoptera ritoralis, method. Claim 13 In paragraph 11 or 12, the above plant is corn, rice, soybean, or cotton. Claim 14 A plant, part thereof, or habitat comprising a composition according to any one of paragraphs 1 to 7. Claim 15 Use of maltodextrin with a DE of 5 to 50 for stabilizing insecticidal proteins for at least 3 days.

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