Media Compositions for Promoting Bacterial and Fungal Growth

Abstract

Methods and compositions for enhancing or promoting germination of bacterial spores, and yeasts are disclosed herein. The composition of the present invention comprises an extract obtained from banana or any member belonging to the genus Musa that may be used alone or in a growth medium to promote and enhance germination of bacterial spores, growth of bacterial, yeast, and fungal cell cultures.

1 . A method of preparing a plant extract comprising the steps of:
 combining a fruit core from a flowering plant, wherein the fruit core is substantially free of: the fruit peel, the fruit ends, and any discoloration, bruises, microbial growth, environmental stress, or any combinations thereof with distilled water;   blending the fruit and distilled water in a processor to form a smooth mixture;   
       separating the blended fruit and distilled water into a supernatant and a sediment, wherein the sediment is discarded and the supernatant is further processed;
 sterilizing the supernatant; 
 optionally performing a second separation on the sterilized supernatant to remove any separated sediment or debris; and 
 clarifying the sterilized supernatant by one or more techniques, wherein the clarification results in an isolation of a fraction of the supernatant of a desired molecular weight range comprising one or more active components responsible for the promotion or enhancement of germination, growth, viability, yield, metabolite production, or any combinations thereof of the one or more microorganisms grown in a media comprising the filtered supernatant. 
 
     
     
         2 . The method of  claim 1 , wherein the method further comprises the optional steps of:
 processing the extract with the one or more active components by one or more techniques selected from lyophilization, vacuum centrifugation, spray drying, or any combinations thereof; and   performing one or more analytical tests or chemical analysis tests on the plant extract, wherein at least one of the test is selected from the group consisting of sugar profile, moisture content, vitamin A analysis, crude protein estimation, complete mineral analysis, non-protein nitrogen (NPN) equivalent to protein, Brix index, specific gravity, vitamin C, crude fiber analysis, pH, fatty acid composition by GC, and any combinations thereof.   
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein the fraction comprising the active components comprises one or more active components in the molecular weight range of 250-1,000 Daltons. 
     
     
         5 . The method of  claim 1 , wherein the microorganisms comprise gram positive bacteria, gram negative bacteria, lactic acid bacteria, aerobic bacteria, anaerobic, bacteria, yeasts, fungi, or any combinations thereof. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the microorganisms comprise a bacterium selected from one or more bacteria belonging to a genus  Bacillus  selected from the group consisting of  Bacillus thuringiensis, Bacillus coagulans, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis , and any combinations thereof. 
     
     
         8 . The method of  claim 1 , wherein the composition is added to a growth, sporulation, or a fermentation medium to promote or enhance spore germination, growth, viability, yield, metabolite production, or any combinations thereof of the one or more microorganisms, wherein the growth medium comprises nutrient broth (NB), nutrient agar (NA), Luria-Bertani broth (LB), Mueller-Hinton cation-adjusted broth (MH), a sporulation broth, eosin-methylene blue agar (EMB), yeast and mold (YM), blood agar, MacConkey agar, Hektoen enteric agar (HE), mannitol salt agar (MSA), Terrific Broth (TB), xylose lysine deoxycholate (XLD), RPMI-1640 media, molasses-based media, buffered charcoal yeast extract agar, minimal media, or any combinations or modifications thereof. 
     
     
         9 . The method of  claim 1 , wherein the composition is added to the growth medium at a concentration ranging from 0.01%-15%, 0.5%-10%, or 1%-5%. 
     
     
         10 . The method of  claim 1 , wherein the flowering plant is selected from a plant belonging to a family selected from the group consisting of Poaceae, Fabaceae, Musaceae, Solanaceae, Cucurbitaceae, Brassicaceae, Apiaceae, Rutaceae, Rosaceae, and any combinations thereof. 
     
     
         11 - 12 . (canceled) 
     
     
         13 . A method for promoting or enhancing germination, growth, viability, yield, metabolite production, or any combinations thereof of one or more microorganisms comprising the step of:
 providing a fermentation or growth medium for a cultivation or growth of the one or more microorganisms;   adding an inoculum or spores of the one or more microorganisms in need of a promotion or enhancement of germination, growth, viability, yield, metabolite production, or any combinations thereof, wherein the inoculum comprises the one or more microorganisms in a lag phase or an exponential phase of a microbial growth cycle; and   adding a plant extract obtained from one or species belonging to the family Musaceae to the growth medium or the inoculum, wherein the plant extract may be added to the growth medium prior to the addition of the inoculum or the spores, or periodically during an exponential or a stationary phase of the microbial growth cycle.   
     
     
         14 . The method of  claim 13 , wherein the method further comprises the steps of:
 monitoring the growth of the one or more microorganisms through the phases of the microbial growth cycle; and   harvesting the one or more microorganisms when a desired growth, viability, or yield is achieved, a desired level of metabolite production is reached, a death phase of the microbial growth cycle is reached, or any combinations thereof.   
     
     
         15 . The method of  claim 13 , wherein the microorganisms comprise gram positive bacteria, gram negative bacteria, lactic acid bacteria, aerobic bacteria, anaerobic bacteria, yeasts, fungi, or any combinations thereof. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 15 , wherein the microorganisms comprise bacteria selected from one or more bacteria belonging to a genus  Bacillus  selected from the group consisting of  Bacillus thuringiensis, Bacillus coagulans, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis , and any combinations thereof. 
     
     
         18 . The method of  claim 13 , wherein the growth medium comprises nutrient broth (NB), nutrient agar (NA), Luria-Bertani broth (LB), Mueller-Hinton cation-adjusted broth (MH), a sporulation broth, eosin-methylene blue agar (EMB), yeast and mold (YM), blood agar, MacConkey agar, Hektoen enteric agar (HE), mannitol salt agar (MSA), Terrific Broth (TB), xylose lysine deoxycholate (XLD), RPMI-1640 media, minimally-based media, molasses-based media, buffered charcoal yeast extract agar, or any combinations or modifications thereof. 
     
     
         19 . The method of  claim 13 , wherein the extract is added to the growth medium at a concentration ranging from 0.01%-15%, 0.5%-10%, or 1%-5%. 
     
     
         20 . The method of  claim 13 , wherein the extract is obtained from one or more fruits of the plant belonging to the family Musaceae, wherein the fruit comprises bananas, plantains,  Ensete, Musella , and any combinations thereof. 
     
     
         21 . The method of  claim 13 , wherein the extract is made from one or more bananas or plantains by a method comprising the steps of:
 blending the bananas or plantains with distilled water in a processor to form a smooth mixture;   centrifuging the mixture to separate a supernatant and a sediment, wherein the sediment is discarded and the supernatant is taken for further processing;   sterilizing the supernatant;   performing a second centrifugation if necessary on the sterilized supernatant to remove any separated sediment or debris; and   purifying the sterilized supernatant by a membrane filtration process, wherein the membrane filtration results in an isolation of a fraction of the supernatant of a desired molecular weight range comprising one or more active components responsible for the promotion or enhancement of the germination, the growth, the viability, the yield, the metabolite production, or any combinations thereof of the one or more microorganisms.   
     
     
         22 . The method of  claim 21 , wherein the method further comprises the optional step of processing the extract with the one or more active components by one or more techniques selected from lyophilization, vacuum centrifugation, spray drying, or any combinations thereof. 
     
     
         23 . The method of  claim 21 , wherein the fraction comprising the active components comprises one or more active components in the molecular weight range of 250-1,000 Daltons. 
     
     
         24 . The method of  claim 21 , wherein the membrane filtration process is performed using a flat-sheet or a hollow fiber membrane, tubular membrane, spiral wound, hollow fiber, pressurized, immersed, or ceramic filtration system. 
     
     
         25 - 27 . (canceled) 
     
     
         28 . A composition for promoting or enhancing germination, growth, viability, yield, metabolite production, or any combinations thereof of one or more microorganisms comprising an extract obtained from one or more flowering plants, made by a method comprising:
 combining a fruit core from a flowering plant, wherein the fruit core is substantially free of: the fruit peel, the fruit ends, and any discoloration, bruises, microbial growth, environmental stress, or any combinations thereof with distilled water;   blending the fruit and distilled water in a processor to form a smooth mixture;   separating the blended fruit and distilled water into a supernatant and a sediment, wherein the sediment is discarded and the supernatant is further processed;   sterilizing the supernatant;   optionally performing a second separation on the sterilized supernatant to remove any separated sediment or debris; and   filtering the sterilized supernatant by a flat sheet or hollow fiber membrane filtration process, wherein the membrane filtration results in an isolation of a fraction of the supernatant of a desired molecular weight range comprising one or more active components responsible for the promotion or enhancement of germination, growth, viability, yield, metabolite production, or any combinations thereof of the one or more microorganisms grown in a media comprising the filtered supernatant.   
     
     
         29 . The composition of  claim 28 , wherein the flowering plant is selected from a plant belonging to a family selected from the group consisting of Poaceae, Fabaceae, Musaceae, Solanaceae, Cucurbitaceae, Brassicaceae, Apiaceae, Rutaceae, Rosaceae, and any combinations thereof. 
     
     
         30 . (canceled) 
     
     
         31 . The composition of  claim 28 , wherein the extract is obtained from a fruit of the flowering plant of the family Musaceae, wherein the fruit is selected from bananas, plantains,  Ensete, Musella , and any combinations thereof. 
     
     
         32 . The composition of  claim 28 , wherein the microorganisms comprise gram positive bacteria, gram negative bacteria, lactic acid bacteria, aerobic bacteria, anaerobic bacteria, yeasts, fungi, or any combinations thereof. 
     
     
         33 . (canceled) 
     
     
         34 . The composition of  claim 32 , wherein the microorganisms comprise a bacteria selected from one or more bacteria belonging to a genus  Bacillus  selected from the group consisting of  Bacillus thuringiensis, Bacillus coagulans, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis , and any combinations thereof. 
     
     
         35 . The composition of  claim 28 , wherein the composition is added to a growth, sporulation, or a fermentation medium to promote or enhance spore germination, growth, viability, yield, metabolite production, or any combinations thereof of the one or more microorganisms, wherein the growth medium comprises nutrient broth (NB), nutrient agar (NA), Luria-Bertani broth (LB), Mueller-Hinton cation-adjusted broth (MH), a sporulation broth, eosin-methylene blue agar (EMB), yeast and mold (YM), blood agar, MacConkey agar, Hektoen enteric agar (HE), mannitol salt agar (MSA), Terrific Broth (TB), xylose lysine deoxycholate (XLD), RPMI-1640 media, minimally-based media, molasses-based media, buffered charcoal yeast extract agar, or any combinations or modifications thereof. 
     
     
         36 . The composition of  claim 28 , wherein the composition is added to the growth medium at a concentration ranging from 0.01%-15%, 0.5%-10%, or 1%-5%. 
     
     
         37 . The composition of  claim 28 , wherein the extract is made by a method comprising the steps of:
 blending the fruit with distilled water in a processor to form a smooth mixture;   centrifuging the mixture to separate a supernatant and a sediment, wherein the sediment is discarded and the supernatant is taken for further processing;   sterilizing the supernatant;   performing a second centrifugation if necessary on the sterilized supernatant to remove any separated sediment or debris; and   purifying the sterilized supernatant by a membrane filtration process, wherein the membrane filtration results in an isolation of a fraction of the supernatant of a desired molecular weight range comprising one or more active components responsible for the promotion or enhancement of the germination, the growth, the viability, the yield, the metabolite production, or any combinations thereof of the one or more microorganisms.   
     
     
         38 . The composition of  claim 37 , wherein the method further comprises the optional step of processing the extract with the one or more active components by one or more techniques selected from lyophilization, vacuum centrifugation, spray drying, or any combinations thereof. 
     
     
         39 . The composition of  claim 37 , wherein the fraction comprising the active components comprises one or more active components in the molecular weight range of 250-1,000 Daltons. 
     
     
         40 - 49 . (canceled)