Antimicrobial, antibacterial and spore germination inhibiting activity from an avocado extract enriched in bioactive compounds

Abstract

The present disclosure relates to extracts from Persea sp. (avocado) enriched in bioactive compounds which can be used as antimicrobial, antibacterial or spore germination inhibiting agents, the process for obtaining the extracts, acetogenins and isolated molecules and methods for using the extracts enriched in bioactive compounds for providing antimicrobial, antibacterial or spore germination inhibiting effect.

What is claimed is: 
     
         1 . A process to obtain an extract enriched in naturally occurring acetogenins from  Persea  sp. having a bacterial spore germination inhibiting effect, which has a starting point a raw extract of the dried or fresh seeds, and/or other  Persea  spp. tissue such as mesocarp, peel, leafstalks, branches or leaves, and which comprises the following steps:
 d) Partitioning of the raw extract serving as an starting point, into a two-phase solvent system to obtain a phase with a high content of acetogenins, further evaporated or concentrated to obtain an extract with a high content of acetogenins;   e) Fractionating the extract with a high content of acetogenins obtained in step a) by Fast or High Performance Centrifugal Partition Chromatography (FCPC or HPCPC) or Countercurrent chromatography (CCC) based on their corresponding partition coefficient, to obtain fractions with higher concentration of acetogenins presenting bacterial spore germination inhibiting effect and separate them from fractions comprising contaminants;   f) Recovering and mixing of the fractions comprising acetogenins with bacterial spore germination inhibiting effect obtained in step b), and concentrating them to finally obtain an extract enriched in naturally occurring acetogenins from  Persea  sp. having bacterial spore germination inhibiting effect.   
     
     
         2 . The process of  claim 1 ; wherein the two-phase solvent system said in step a) comprises: at least one polar solvents selected from the group:
 water, C 1 -C 4  alcohol (e.g. ethanol, isopropanol, methanol), dimethyl sulfoxide, tetrahydrofuran, acetone, acetonitrile; and   at least one non-polar solvents selected from the group:   hexanes, heptanes, ethyl ether, ethyl acetate, petroleum ether, butyl alcohol, chloroform, toluene, methyl tert-butyl ether, methyl isobutyl ketone and mixtures therein.   
     
     
         3 . The process of  claim 1 ; wherein the fractionation by FCPC, HPCPC or CCC said in step b) is carried out by use of a two-phase solvent system comprising at least one of the following combinations, all of them at different ratios, alone or in parallel:
 methanol:heptanes, water:hexane, water:butanol, methyl tert-butyl ether: acetonitrile:water, heptanes:ethyl acetate: acetonitril, heptanes:ethyl acetate:methanol:water.   
     
     
         4 . The process of  claim 1 ; wherein the recovered fractions comprising acetogenins with bacterial spore germination inhibiting effect said in step c) have a partition coefficient value lower than 0.5, and preferably in the in the range of between 0.19 to 0.35, when fresh seeds are used and FCPC, HPCPC or CCC is carried out with a heptane:methanol two-phase solvent system and heptane as initial stationary phase. 
     
     
         5 . An extract enriched in acetogenins from  Persea  spp; wherein the extract is characterized by having bacterial spore germination inhibiting effect. 
     
     
         6 . The extract of  claim 5 ; wherein the spore germination inhibiting effect of said extract can be applied to bacterial spores from the genera which includes, but is not limited to  Clostridium, Bacillus, Alicyclobacillus.    
     
     
         7 . The extract of  claims 5  and  6 ; wherein the extract inhibits bacterial spores from at least one bacteria selected from  Clostridium botulinum, Clostridium perfringens, Clostridium difficile, Bacillus anthracis, Bacillus cereus, Bacillus subtilis, Bacillus lichniformis, Alicyclobacillus acidoterrestris, Alicyclobacillus acidiphilus.    
     
     
         8 . The extract of  claim 5 ; wherein the extract comprises at least one compound characterized by the formula (I) 
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is an acetyl group; 
         R 2  is hydrogen or a hydroxy protecting group; and 
         R 3  is an is an alkenyl group with at least one carbon-carbon double bond; and/or compounds of formula (II) 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is an acetyl group; 
         R 2  and R 4  hydrogen or a hydroxy protecting group; and 
         R 3  is an alkenyl group with at least one carbon-carbon double bond. 
       
     
     
         9 . The extract of  claim 8 ; wherein all the stereoisomeric forms (R) and (S), and the double bonds in cis (Z) or trans (E) configuration are included. 
     
     
         10 . The extract of  claims 8  and  9 ; wherein the extract comprises at least one compound preferably characterized by the formula (I), and wherein there is at least one double bond with trans configuration at the C-5 and C-6 position of the compound. 
     
     
         11 . The extract of  claims 8  and  9 ; wherein the extract comprises at least one compound preferably characterized by the formula (I), and wherein there is at least one double bond with trans configuration at the C-16 and C-17 position of the compound. 
     
     
         12 . The extract of  claims 8  and  9 ; wherein the extract comprises at least one compound preferably characterized by the formula 
       
         
           
           
               
               
           
         
       
     
     
         13 . Use of the extract of  claims 5  to  12  to make a composition or product for providing an spore germination inhibiting effect; wherein the composition or product is selected from the group consisting of:
 a pharmaceutical composition, comprising the extract and a pharmaceutically acceptable carrier; wherein the pharmaceutical composition is suitable for one or more of the following administration vias: 
 oral, dermal, parenteral, nasal, ophthalmical, optical, sublingual, rectal, gastrical or vaginal; 
 a food additive composition comprising the extract and a food grade acceptable carrier, suitable for inclusion into food products; wherein the food product is selected from one of more of the following: 
 fish, crustaceans, fish substitutes, crustacean substitutes, meat, meat substitutes, poultry products, vegetables, greens, sauces, emulsions, beverages, juices, wines, beers, dairy products, egg-based products, jams, jellies, grain-based products, baked goods and confectionary products; 
 a personal care products; wherein the personal care composition is selected from one or more of the following: 
 creams, gels, powders, lotions, sunscreens, lipstick, body wash, herbal extracts, and formulations that support the growth of bacteria; and 
 a cleaning composition; wherein the cleaning composition is suitable for application to one of the following: 
 counter tops, doors, windows, handles, surgical equipment, medical tools, and contact surfaces that can contaminate humans or animals. 
 
     
     
         14 . Composition or product comprising at least one of the extracts of  claims 5  to  12  characterized by having an inhibitory effect in spore germination. 
     
     
         15 . Composition or product comprising at least one of the extracts of  claim 10  characterized by having an inhibitory effect over the genera  Listeria  at storage temperatures in the range of 0 to 10° C. 
     
     
         16 . Composition or product comprising at least one of the extracts of  claims 11  and  12  characterized by having an antibacterial, antifungical, antiviral, anti-yeast, and in spore germination inhibitory effect. 
     
     
         17 . Use of the compositions or products of  claims 14  to  16 , wherein the composition or product is selected from the group consisting of:
 a pharmaceutical composition, comprising the extract and a pharmaceutically acceptable carrier; wherein the pharmaceutical composition is suitable for one or more of the following administration vias: 
 oral, dermal, parenteral, nasal, ophthalmical, optical, sublingual, rectal, gastrical or vaginal; 
 a food additive composition comprising the extract and a food grade acceptable carrier, suitable for inclusion into food products; wherein the food product is selected from one of more of the following: 
 fish, crustaceans, fish substitutes, crustacean substitutes, meat, meat substitutes, poultry products, vegetables, greens, sauces, emulsions, beverages, juices, wines, beers, dairy products, egg-based products, jams, jellies, grain-based products, baked goods and confectionary products; 
 a personal care products; wherein the personal care composition is selected from one or more of the following: 
 creams, gels, powders, lotions, sunscreens, lipstick, body wash, herbal extracts, and formulations that support the growth of bacteria; and 
 a cleaning composition; wherein the cleaning composition is suitable for application to one of the following: 
 counter tops, doors, windows, handles, surgical equipment, medical tools, and contact surfaces that can contaminate humans or animals.