Process for preparation of natural and organic potassium sorbate

Abstract

PROCESS FOR PREPARATION OF NATURAL AND ORGANIC POTASSIUM SORBATE The present invention relates to the process for production of natural and organic potassium sorbate. The natural and organic potassium sorbate is produced through microbial fermentation of natural carbohydrate source obtained from organic raw material tuber crop cassava followed by downstream processing steps of filtration, carbon treatment, evaporation, centrifugation and spin flash drying. The fermentation was carried out utilizing the Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705, which were modified by the way of strain improvement through media optimization, filtration, carbon treatment, evaporation and centrifugation, the pellet material received was further dried in spin flash dryer resulting in finished product in powder form as potassium sorbate with sorbic acid (70-72%), other organic acids (10%),bioavailable potash (12%) and moisture (<5%). The natural and organic potassium sorbate powder produced has antimicrobial activity and addition of 0.1 to 0.5 % of the said natural preservative in food preparation of different food products like cheese and paneer, pickles, meat products, beverages like wines, canned food items, storage food items and bakery preparations enhances the shelf life of the product. It kills variety of food pathogens viz. Listeria sp., Salmonella sp., and Shigella sp in breads, paneer and pickles and on crops as foliar and drenching application to control soil borne phytopathogens.

Description

PROCESS FOR PREPARATION OF NATURAL AND ORGANICPOTASSIUM SORBATEFIELD OF INVENTION

[0001] The present invention relates to the process for preparation of natural and organic potassium sorbate, having profound broad spectrum antimicrobial properties. The invention illustrates the production of from vegan sources having broad spectrum antimicrobial properties. The process for preparation was carried out by microbial fermentation process utilizing non edible tuber crop cassava based glucose using microbial consortia. The natural and organic potassium sorbate was found to be antimicrobial agent against broad categories of food borne pathogens causing spoilage of food stuffs.BACKGROUND OF THE INVENTION

[0002] The preservation, safety, and freshness of the goods in our system of global food supply depend heavily on preservatives. Because of contemporary (and traditional) methods of food preservation, many of the foods we eat today are possible. Food manufacturers are able to transport their products across the nation and the world without compromising the quality or safety of the food because preservatives and preservation procedures stop meals from fast rotting and oxidizing. Microbes and oxidation are two important factors that cause food to go bad quickly. Unwanted bacteria, fungus, and yeasts can develop in our food goods and cause spoilage. If consumed by people, these microbes can seriously injure them since they feed on the nutrients in food. Bacteria like listeria and botulinum can enter our foods without preservatives if are introduced through food items. Food preservatives are essential in halting food degradation because they guard against the growth of mold, yeast, deadly botulinum, and other organisms that can lead to food poisoning. Preservatives also make food more convenient, extend shelf life, and decrease food waste. All preservatives are under FDA regulation,and the Food Safety and Inspection Service is also in charge of overseeing the safety of food additives used in meat, poultry, and egg products. The FDA prohibits using preservatives in a way that hides damage or deficiency, makes the food appear better than it is, or negatively impacts the food's nutritional content. The U.S. Code of Federal Regulations contains a list of food additives that are permitted for use as preservatives. In the processed food, sodium nitrite / nitrate present significant amount. Large consumption of processed meats has been associated in studies to a higher risk of colorectal cancer. Sulfites and sodium benzoate seem to be harmless for the majority of people, although they can have negative effects on some. It is also reported that, sodium benzoate and artificial food colorings may make young toddlers more hyperactive.The National Toxicology Program classifies butylated hydroxy anisole, or BHA, as "reasonably anticipated to be a human carcinogen," but the FDA views it as a GRAS substance at small doses. However, despite the fact that butylated hydroxy toluene, or BHT, has been outlawed in several nations, there is still no proof that it is harmful. Exclusively consuming high fat, high sugar products that contain preservatives will have negative long-term health consequences. However, we have not yet determined that these health impacts are related to the use of chemical preservatives within recommended quantities or are the consequences of an overall unhealthy lifestyle. Research is ongoing regarding specific preservative ingredients and their health impacts. Researchers and the U.S. FDA take food safety seriously and strive to ensure all foods and food products are safe for everyone to consume. Thus, researchers work with food regulatory agencies to share essential preservative discoveries so agencies can modify preservation regulations and guidelines based on science.

[0003] The majority of our dietary calories and around half of our protein needs come from bakery goods and cereals, which are important sources of nutrients in our diet. The use of natural, whole grains and other components has increased the importance of bakery products. Additionally, as starch is the primary chemicalcomponent of bakery goods, they are regarded as a source of carbohydrates (Jageethadevi et al., 2012). Products from bakeries are prone to spoiling issues. These include microbiological, chemical, and physical deterioration. Since water activity is the primary cause of bakery product deterioration, mold growth and other microbiological degradation represent the primary economic significance of bakery products. For bakeries, mold deterioration is a significant and expensive issue (Saranraj and Geetha, 2012).

[0004] Preservatives are primarily responsible for the stability of bakery products against the attack of fungi. Preservatives aid in reducing or preventing food decay brought on by microorganisms. A wider range of products may be kept in stock and at home because of longer shelf lives. According to Sofos and Busta (1991), chemical preservatives can inhibit the metabolism of molds, denature the cell's protein, or physically harm the cell membrane in addition to stopping their growth. Propionic and sorbic acid, or their salts, are two of these preservatives that have been proven to lengthen the shelf life of bread products.

[0005] Sorbic acid and its product salt are essentially safe to humans when used in ordinary amounts and have anti-microbial properties. As a result, these substances can be added to meals. As a food preservative and freshness-preserving agent, potassium sorbate inhibits mold growth, yeast growth, and aerobic bacterial growth as well as the growth and breeding of harmful microorganisms like Clostridium botulinum, Staphylococcus and Salmonella.PRIOR ARTS

[0006] United States Patent US3320307A explains the process for the production of potassium sorbate where the production of an alkali metal sorbate comprises reacting a solution of sorbic acid in acetone with an alkali metal carbonate.

[0007] US3173948A (1963) patent describes the preparation of crystalline potassium sorbate from a basic aqueous solution.

[0008] CN1634844A (2006) Chinese patent explains the preparation technology of potassium sorbate comprising seven steps with acetic acid as starting material using chemical synthesis method.

[0009] However, none of the prior arts describe the present invention. The prior arts emphasize the preparation of potassium sorbate from chemical synthesis. The prior arts indicates the use of chemicals which are not environment friendly and may cause toxicity in the environment leaving toxic residues, impurities and non- biodegradable waste in the environment. There is no conventional novel, biological, natural and fermentation process available for production of sorbic acid and potassium sorbate. Therefore, the present invention is a process from pilot scale to commercial production to produce natural sorbic acid and potassium sorbate. The present process is unique, biological natural, environment friendly and leaves no toxic residues in the environment. Thus, the present process is more effective in comparison to the processes mentioned in prior arts.SUMMARY OF THE INVENTION

[0010] The present invention is a process for the preparation of natural and organic potassium sorbate, which is effective in inhibiting the growth of food spoilage pathogens and is helpful in extending the shelf life of food items viz. meat products, paneer, pickle and bread samples. The present invention is also found effective in controlling phyto-pathogens and insects / pests in agriculture.

[0011] It is a promising and an effective broad spectrum antimicrobial agent / preservative based on vegan source and clean label product, an effective shelf life enhancer in preserving almost all food items in comparison to the chemical preservatives. The present invention utilizes the natural and organicfermentation process for preparation of natural potassium sorbate. The process of preparation of natural potassium sorbate emphasizes that, the finished preparation, natural and organic potassium sorbate which doesn’t have any toxic effects on gut health and is found as natural alternative preservative in comparison to chemical preservatives.

[0012] In one of the embodiment, the present invention relates to the process for preparation of natural and organic potassium sorbate by microbial fermentation process from natural carbohydrate source like cassava followed by downstream processing steps filtration, carbon treatment, evaporation, centrifugation and spin flash drying. After drying, the product is in powder form with sorbic acid (70- 72%), other organic acids (10%), bioavailable potash (12%) and moisture (<5%).

[0013] In another embodiment of the present invention, natural and organic potassium sorbate is produced by biological fermentation utilizing consortia of microbes namely Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705. The microbes are grown in suitable designed media for the production of sorbic acid and other organic acids in proper proportions. These bacterial strains were originally procured from National Collection of Industrial Microorganisms (NCIM) at National Chemical Laboratory, Pune and were modified by way of strain improvement methods by media optimization. Different media compositions with varying percentage of C / N ratios were prepared and inoculated with microbial cultures used in the study to understand the optimal medium for significant and efficient production of Sorbic acid.

[0014] In another embodiment of the present invention for production of natural and organic potassium sorbate is the production of dextrin (>85% DE) usingliquefaction enzyme and glucose (>95% DE) which was produced using saccharification enzymes catalyzing the flour starch.

[0015] Further to the present invention, fermentation was carried out utilizing the bacterial consortia comprising Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705 in combination at suitable pH with media composition, viz. Cassava based Dextrin solution (500 mL / L) + Cassava based Glucose solution (400 mL / L) + Yeast powder (60 g / L) + Yeast extract powder (20 g / L) + Dipotassium phosphate (20 g / L) + Potassium dihydrogen phosphate (10 g / L) + CSL powder (30 g / L) + Potassium hydroxide adjusted with pH 6.5 - 7.2 at 32±1°C for 15 days with intermittent agitation and aeration duration of 10 days after first 5 days of fermentation process.

[0016] In another embodiment, fermented broth results in production of sorbic acid and other organic acids viz. sorbic acid (8-9%), lactic acid (0.5- 1.0%) and acetic acid (1-1.5%). The pH decrement in the batches was neutralized with potassium hydroxide in order to produce potassium sorbate as the finished product. The broth was harvested with increase in temperature at 90-95°C for 2 hours followed with filtration, carbon treatment, evaporation and centrifugation. The pellet material received was further dried in spin flash dryer resulting in finished product in powder form as natural and organic potassium sorbate.

[0017] The present process for preparation of natural and organic potassium sorbate is unique and was validated by multiple experimental pilot scale (5,000 Liters) and commercial (30,000 Liters and 50,000 Liters) fermenters.

[0018] Another embodiment of the present invention relates to the addition of 0.1shelf life considerably and gives better results in comparison to chemical based potassium sorbate.BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 : Flow chart for preparation of natural and organic potassium sorbate.

[0020] Figure 2: Graph showing the concentration of TRS and Sorbic acid percent in the process samples with respect to time- Depicts the change in concentration of substrate Total Reducing Sugar and the product Sorbic acid for the in-process fermentation samples of collected during different time interval.DETAILED DESCRIPTION OF THE INVENTION

[0021] The invention relates to a process for preparation of natural and organic potassium sorbate via natural and organic fermentation process using natural carbohydrate source cassava using a bacterium consortium comprising of Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705. The microbes are grown in suitable designed media for the production of sorbic acid and other organic acids in proper proportions. These bacterial strains were originally procured from National Collection of Industrial Microorganisms (NCIM) at National Chemical Laboratory, Pune and were modified by way of strain improvement methods by media optimization. The natural preservative preparation is having broad spectrum antimicrobial activity against the food spoilage pathogens deteriorating the food safety.

[0022] In one of the embodiment of the present invention relates to “Preparation of Natural and Organic Potassium Sorbate”, which is a completely organic, natural broad spectrum antimicrobial preservative which is produced by utilizing raw material in the form of grist of non-edible tuber crop cassava. The cassavatubers are dried and pulverized to produce cassava starch, which in then after treated with liquefaction enzymes to produce dextrin (>85% DE).

[0023] In another embodiment of the present invention for production of natural and organic potassium sorbate, the dextrin produced by liquefaction enzyme is further catalyzed by saccharificaton enzymes for production of glucose (>95% DE).

[0024] Further to the present invention, fermentation was carried out utilizing the bacterial consortia comprising Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705 in combination at suitable pH with media composition, viz. Cassava based Dextrin solution (500 mL / L) + Cassava based Glucose solution (400 mL / L) + Yeast powder (60 g / L) + Yeast extract powder (20 g / L) + Dipotassium phosphate (20 g / L) + Potassium dihydrogen phosphate (10 g / L) + CSL powder (30 g / L) + Potassium hydroxide adjusted with pH 6.5 - 7.2 at 32±1°C for 15 days with intermittent agitation and aeration duration of 10 days after first 5 days of fermentation process.

[0025] In another embodiment, fermented broth results in production of sorbic acid and other organic acids viz. sorbic acid (8-9%), lactic acid (0.5- 1.0%) and acetic acid (1-1.5%). The pH decrement in the batches was neutralized with potassium hydroxide in order to produce potassium sorbate as the finished product. The broth was harvested with increase in temperature at 90-95°C for 2 hours followed with filtration, carbon treatment, evaporation and centrifugation. The pellet material received was further dried in spin flash dryer resulting in finished product in powder form as natural and organic potassium sorbate.

[0026] The present invention related to the process for preparation of natural and organic potassium sorbate by microbial fermentation process from natural carbohydrate source like cassava followed by downstream processing steps filtration, carbon treatment, evaporation, centrifugation and spin flash drying. After drying, the product will be in powder form with sorbic acid (70-72%), other organic acids (10%), bioavailable potash (12%) and moisture (<5%).

[0027] Another embodiment of the present invention relates to the addition of 0.1 - 0.5% of Natural and organic potassium sorbate to food products extends the shelf life considerably and gives better results in comparison to chemical based potassium sorbate.

[0028] The present invention is being used as food preservative in different food products like cheese and paneer, pickles, meat products, beverages like wines, canned food items, storage food items and bakery preparations. The present invention describes that the invention can be utilized in agricultural formulations for foliar and soil-based applications for eliminating the bacterial and fungal pathogens invading the crops. The present invention is effective and significant preservative agent, utilized as broad-spectrum antimicrobial activity against the food borne pathogens and the pathogenic strains which causes food spoilage like Listeria, Salmonella and Shigella.

[0029] The natural and organic potassium sorbate produced in the present invention when used for preservation in meat products extends the shelf life till 28 days in comparison to set treated with chemical-based potassium sorbate which was having only 7 days’ time period. Further, it was found that natural and organic potassium sorbate extends the shelf life of paneer for 38 days in comparison chemical-based potassium sorbate which showed only 20 days time period. The pickle samples sets were also treated with natural and organic potassium sorbate and chemical-based potassium sorbate separately. The results revealed that, shelf life of set of pickles treated with natural and organic potassium sorbate were found to have 182 days in comparison to 94 days (treated with chemical-based potassium sorbate). The bread doughs were prepared using natural and organic potassium sorbate and chemical-based potassium sorbate. The shelf life of bread sets treated with natural and organic potassium sorbate was found to be 38 days in comparison to bread sets treated with chemical potassium sorbate which showed only 25 days shelf life.Experimental Details & ResultsEXAMPLE 1(i) Upstream Process Parameters

[0030] The present invention, natural and organic potassium sorbate is produced by biological fermentation utilizing consortia of microbes namely Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705. The fermentation process was carried out using microbial consortia comprising of Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705 in media with composition as mentioned below at suitable pH 6.5 - 7.2 adjusted with potassium hydroxide and temperature of 32+1 °C.

[0031] The microbes were grown in suitable designed media for the production of sorbic acid and other organic acids in proper proportions. These bacterial strains were originally procured from National Collection of Industrial Microorganisms (NCIM) at National Chemical Laboratory, Pune which were modified by way of strain improvement methods using media optimization changing C / N ratios in the growth medium (Table 1-4). The best modified medium that showed efficient production of sorbic acid and reflected effective growth of bacterial strains was utilized as the optimal growth medium for sorbic acid production (Table 4).Table 1: Media composition - 1Media composition for production of sorbic acid and effective growth of microbesTable 2: Media composition - 2Media composition for production of sorbic acid and effective growth of microbesTable 3: Media composition - 3Media composition for production of sorbic acid and effective growth of microbesTable 4: Media composition - 4Optimal Media composition for effective production of sorbic acid and effective growth of microbes

[0032] Media was heat sterilized at 121°C and 15 psi pressure for 25 min and allowed to cool down to room temperature. The pre- sterilized fermentation medium in the fermenter was inoculated with 10% of inoculum of microbial consortia comprising of Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705 from 48 h grown static flask cultures. Temperature and pH were set at 32°C and 6.5 - 7.2respectively with an agitation of 100 rpm. The pH was maintained and the produced organic acids were neutralized with periodical addition of sterile potassium hydroxide slurry.EXAMPLE 2(ii) In-process Monitoring of Total Reducing Sugars (TRS) and Product Yield

[0033] To 25 mL of fermented broth sample, 5 mL of HC1 and 50 mL of water was added and heated for about 1 hour. After 1 hour, it was cooled to bearable temperature. Sample was poured in a 250 mL volumetric flask, made up to 250 mL and filtered.

[0034] 10 mL of filtered sample was taken and made up to 100 mL in a volumetric flask. 5 mL of Fehling solution A and 5 mL of Fehling solution B were taken in a conical flask. 50 mL of distilled water was added to it and allowed to boil in a heating mantle. When boiling, 2 - 3 drops of methylene blue indicator was added. Above prepared sample was then titrated until brick red color has been reached.0.05 X 100 X 100Total Reducing Sugar % =Titration volume X Sample volume

[0035] The TRS was initially found to be 10.5 % during the start of the fermentation process. After inoculation with microbial consortia the fermentation process got initiated and gradually the reducing sugars got deteriorated and were getting converted to product. The decrease in TRS percentage with respect to time is depicted in Figure 2 and was found that complete utilization of TRS happened within 120 hours.

[0036] Yield of the organic acids formed were analyzed in the in-process samples as well as finished product sample by High Performance Liquid Chromatography based method. 0.1 g of test sample was dissolved in 100 mL of HPLC water. Degassing was performed with an ultrasonicator to prepare the test sample.Further, the sample was filtered with a sterile 0.2 pm PTFE filter (Axiva 200050 RI, AXIVA Sichem Biotech Pvt. Ltd., India). The samples were analyzed with reference to analytic reference standards of respective organic acids.

[0037] Further, samples were analyzed by injecting 20 pL of the prepared samples into the HPLC (Shimadzu LC2030CHT) system. Organic acids column (250 X 4.6 mm) was used by maintaining column temperature at 30°C against 8 mM sulphuric acid in water mobile phase. The flowrate was maintained at 0.5 mL / min while the total run time was 35 min. Detection was performed through UV Vis at 215 nm.

[0038] The standards were injected using the same conditions at concentrations ranging from 2 mM to 20 mM to create a standard curve. Using a spreadsheet application, the peak areas of the standards against their concentration were plotted. Further the slope and intercept of the least square regression line were determined. Checked the line for linearity and discarded the low or high points that are not linear. The test samples were ensured that their absorbance falls within the range of the linear standard concentrations.

[0039] Using the Shimadzu Lab Solutions software, the concentration of respective organic acids in a test sample were determined with reference to the standard calibration curve of respective organic acids in terms of difference of sample peak area and the intercept of gradient of organic acids plotted against the slope of standard curve for each of the individual organic acids.

[0040] The concentration of the product Sorbic acid that was formed during the fermentation process at different interval of time was determined and is shown in Figure 2.EXAMPLE 3(iii) Downstream Processing and Product Recovery

[0041] Complete utilization of reducing sugars was achieved within 120 h of fermentation. Further filtration was performed using 10 micron cloth filters in aplate and frame filtration assembly. The filtered product was collected in a collection tank and carbon treatment followed by carbon filtration was conducted to reduce the color of the filtrate liquid. Further, it was vacuum evaporated at 65°C temperature under vacuum to increase the concentration of the filtrate. The potassium fortified liquid product was then centrifuged and the pellet material obtained was further spin flash dried at a temperature of 230 - 250°C to produce powder form of natural and organic potassium sorbate. The naturally produced potassium sorbate powder contains sorbic acid (70-72%), other organic acids (10%), bioavailable potash (12%) and moisture (<5%). The flow chart for preparation of natural and organic potassium sorbate has been shown in Figure 1.EXAMPLE 4(iv) a) Shelf life studies of various food products treated with Natural and Organic Potassium sorbate in comparison to chemical potassium sorbate b) Fungal pathogens; pests control and Vigor index measurement in Okra plants treated with Natural and Organic potassium sorbate in comparison to chemical potassium sorbate

[0042] The shelf life studies were conducted for various food products with preservatives Natural and Organic Potassium sorbate and chemical potassium sorbate and results were compared as follows. In the meat samples sets, treated with natural and organic potassium sorbate Vs chemical based inorganic potassium sorbate, the results revealed that, the set of meat samples treated with natural and organic potassium sorbate, the meat samples were found to be stable for a period of 28 days (microbial load-3.0 cfu / g) while the sets treated with chemical based potassium sorbate was found to be stable for 7 days only (having high microbial load -100 cfu / g). The preservation using Natural and Organic Potassium sorbate in meat products extends the shelf life till 28 days in comparison to set treated with chemical-based potassium sorbate which was having only 7 days’ time period (Table 5).

[0043] In the paneer samples, treated with natural and organic potassium sorbate Vs natural and organic calcium propionate, the results revealed that, the paneer samples sustained for 38 days treated with natural and organic potassium sorbate (microbial load -2 cfu / g) while the sets treated with natural and organic calcium propionate, the paneer sample was found to be stable for 20 days (microbial load- 15 cfu / g). Therefore, it was found that, natural and organic potassium sorbate extends the shelf life of paneer for 38 days in comparison chemical-based potassium sorbate which showed only 20 days’ time period (Table 6).

[0044] In the pickles samples, treated with natural and organic potassium sorbate Vs chemical potassium sorbate Vs natural and organic calcium propionate, the stability of pickle sample was found to be stable for 182 days (microbial load -10 cfu / g) treated with natural and organic potassium sorbate while the sets treated with chemical potassium sorbate, the stability of the pickle samples sustained for 94 days (microbial load- 23 cfu / g) while the sets treated with natural and organic calcium propionate, the shelf life of pickles remained for 45 days (microbial load-30 cfu / g). The pickle samples sets were also treated with natural and organic potassium sorbate and chemical-based potassium sorbate separately. The results revealed that, shelf life of set of pickles treated with natural and organic potassium sorbate were found to have 182 days in comparison to 94 days treated with chemical -based potassium sorbate (Table 7). The bread doughs were prepared using natural and organic potassium sorbate and chemical-based potassium sorbate.

[0045] In the bread samples, treated with natural and organic potassium sorbate Vs chemical potassium sorbate Vs natural and organic calcium propionate, the stability of bread sample was found to be stable for 38 days (microbial load -3 cfu / g) treated with natural and organic potassium sorbate while the sets treated with chemical potassium sorbate, the stability of the bread samples sustained for 25 days (microbial load- 15 cfu / g) while the setstreated with natural and organic calcium propionate, the shelf life of bread samples remained for 15 days (microbial load-20 cfu / g). Therefore, the shelf life of bread sets treated with natural and organic potassium sorbate was found to be 38 days in comparison to bread sets treated with chemical potassium sorbate which showed 25 days of shelf life (Table 8).

[0046] The studies were also conducted in agriculture fields of Chemical potassium sorbate and Natural and Organic potassium sorbate on Okra crop at a concentration of 1.5 ml / Liter to reduce the pest attack and fungal pathogens infestations. It was observed that, fungal pathogens gets reduced to 5.0 cfu / g in Natural and Organic Potassium sorbate treated crop sets in comparison to 20.0 cfu / g in Chemical potassium sorbate treated crop sets. It was also observed that, Natural and Organic Potassium Sorbate reduces the pest population at dosage of 1.5 ml / L to 10.0 in comparison to 32.0 in case of Chemical potassium sorbate. The overall vigor index of Natural and Organic Potassium Sorbate treated crops at dosage of 1.5 ml / Liter showed higher values (89.0) in comparison to Chemical potassium sorbate which showed vigor index as 28.0 (Table 9).Table 5: Shelf life studies of meat samples treated with Natural and Organic Potassium sorbate Vs Chemical based Potassium sorbate to study preservative potential**a) Spore count of Listeria sp. - 0- 10.0 Cfu / g was recorded as acceptable limits; b) Spore count of Listeria sp. - 10.0-100.0 Cfu / g was recorded as non- satisfactorily; c) Spore count of Listeria sp. - >100.0 Cfu / g was found non acceptable [*as per standard FSSAI guidelines, October, 2021]Table 6: Shelf-life studies of paneer samples treated with Natural and Organic Potassium sorbate Vs Chemical based potassium sorbate to study preservative potential**, Most effective treatment**a) Spore count of Listeria sp. - 0- 10.0 Cfu / g was recorded as acceptable limits; b) Spore count of Listeria sp. - 10.0-100.0 Cfu / g was recorded as non- satisfactorily; c) Spore count of Listeria sp. - >100.0 Cfu / g was found non acceptable [*as per standard FSSAI guidelines, October, 2021]Table 7: Shelf-life studies of pickle samples treated with Natural and Organic Potassium sorbate Vs Chemical potassium sorbate Vs Natural Calcium propionate to study preservative potential**, Most effective treatment**a) Spore count of Listeria sp. 0- 10.0 Cfu / g was recorded as acceptable limits; b) Spore count of Listeria sp. - 10.0-100.0 Cfu / g was recorded as non- satisfactorily; c) Spore count of Listeria sp. - >100.0 Cfu / g was found non acceptable [*as per standard FSSAI guidelines, October, 2021]Table 8: Shelf-life studies of bread samples treated with Natural and Organic Potassium sorbate Vs Chemical potassium sorbate to study preservative potential**, Most effective treatment**a) Spore count of Listeria sp. - 0- 10.0 Cfu / g was recorded as acceptable limits; b) Spore count of Listeria sp. - 10.0-100.0 Cfu / g was recorded as non- satisfactorily; c) Spore count of Listeria sp. - >100.0 Cfu / g was found non acceptable [*as per standard FSSAI guidelines, October, 2021]Table 9: Effect of Natural and organic potassium sorbate Vs Chemical based potassium sorbate for control of fungal pathogens and pests and effect on Vigor index in Okra crop**, Most effective treatmentIndustrial Applicability

[0047] The potassium sorbate powder produced in the present invention has antimicrobial activity and addition of 0.1 to 0.5 % of the said natural preservative in food preparation of different food products like cheese and paneer, pickles,meat products, beverages like wines, canned food items, storage food items and bakery preparations enhances the shelf life of the product. It kills broad variety of food pathogens viz. Listeria sp., Salmonella sp., and Shigella sp. in breads, paneer and pickles and on crops as foliar and drenching application to control soil borne phytogens like bacterial and fungal pathogens.REFERENCES1. Jageethadevi, A., P. Saranraj and N. Ramya. 2012. Inhibitory effect of chemical preservatives and organic acids on the growth and organic acids on the growth of bacterial pathogens in poultry chicken. Asian Journal of Biochemical and Pharmaceutical Research, 1 (2): 1 - 9.2. Saranraj, P and M. Geetha. 2012. Microbial spoilage of Bakery products and its control by preservatives. International Journal of Pharmaceutical and Biological Archives, 3 (1): 204 - 214.3. Sofos J. N. and Busta, F, 1991. Antimicrobial activity of sorbate. Journal of Food Protection, 44: 61 4 - 621.4. Chinese application no. CN1634844A. Retrieved from https: / / patents.google.com / patent / CN1634844A5. Kerr Frank Ernest (1963) United States application no. US3320307A. Retrieved from https: / / patents.google.com / patent / US3320307A.6. Probst Otto, Oehme Horst (1963) United States application no.US3173948A. Retrieved from https: / / patents.google.com / patent / US3173948A.

Claims

AIM:

1. A process for production of natural and organic potassium sorbate produced by microbial fermentation of natural carbohydrate source obtained from organic raw material tuber crop cassava followed by downstream processing steps of filtration, carbon treatment, evaporation, centrifugation and spin flash drying; wherein the microbial source used for fermentation process is a bacterial consortium comprising of Lactobacillus delbrueckii NCIM 2365, Bacillus coagulans MTCC 3543 and Lactobacillus acidophilus NCIM 5705 which were modified by the way of strain improvement through media optimization and fermentation is carried out on a medium comprising of the following composition: Cassava based Dextrin solution (500 mL / L), Cassava based Glucose solution (400 mL / L), Yeast powder (60 g / L), Yeast extract powder (20 g / L), Dipotassium phosphate (20 g / L), Potassium dihydrogen phosphate (10 g / L), CSL powder (30 g / L) and Potassium hydroxide adjusted for pH 6.5 - 7.2.

2. The process as claimed in claim 1, wherein fermentation is carried out in 50 L stirred tank Stainless Steel fermenters and sequentially scaled up to 30,000 or 50,000 L fermenters in batch mode at temperature 32+1 °C and pH 6.5 - 7.2 and agitation 100 rpm.

3. The process as claimed in claim 2, wherein production batch is terminated between 120 h of fermentation in commercial fermenter.

4. The process as claimed in claim 1, wherein filtration is carried out through 10 micron size cloth filter in a plate and frame filtration assembly followed by carbon treatment and filtration in 10 micron filter cloth, evaporation carried out at 65 °C temperature under vacuum, centrifugation and spin flash drying at temperature 230 - 250°C.

5. The process as claimed in claims 1 to 4, wherein neutralization of the organic acid produced through fermentation is done by dosing ofpotassium source and thereby providing potassium fortification in the product.

6. The process claimed in claims 1 to 5 produces a natural and organic potassium sorbate which is in powder form.

7. The natural and organic potassium sorbate as claimed in claim 6, comprises of naturally produced sorbic acid (70-72%), other organic acids (10%), bioavailable potash (12%) and moisture (<5%).

8. The natural and organic potassium sorbate as claimed in claim 5, wherein addition of 0.1 to 0.5 % of the said natural preservative in food preparation of different food products like cheese and paneer, pickles, meat products, beverages like wines, canned food items, storage food items and bakery preparations enhances the shelf life of the product; and dosage of 1.5 ml / Liter of product on Okra plant controls the growth of fungal pathogens, pests population and overall enhancement in vigor index of the crop.

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