Title of Invention	AN IMPROVED PROCESS FOR THE PREPARATION OF PSEUDOBACTIN USEFUL FOR INCREASING THE SHELF LIFE OF STORED FRUITS, VEGETABLES AND TUBERS
Abstract	This invention relates to an improved process for the preparation of pseudobactin useful for increasing the shelf life of stored fruits, vegetables and tubers, which comprises cultivating a novel strain of Pseudomonus sp. having characteristics such as herein described in a conventional iron free medium at pH 7 in a temperature ranging 25-300C for 48 h recovering pseudobactin from culture medium by conventional chromatographic methods such as herein described.

Title of Invention

AN IMPROVED PROCESS FOR THE PREPARATION OF PSEUDOBACTIN USEFUL FOR INCREASING THE SHELF LIFE OF STORED FRUITS, VEGETABLES AND TUBERS

Abstract

This invention relates to an improved process for the preparation of pseudobactin useful for increasing the shelf life of stored fruits, vegetables and tubers, which comprises cultivating a novel strain of Pseudomonus sp. having characteristics such as herein described in a conventional iron free medium at pH 7 in a temperature ranging 25-300C for 48 h recovering pseudobactin from culture medium by conventional chromatographic methods such as herein described.

Full Text	This invention relates to an improved process for the preparation of a pseudobactin formulation useful for increasing the shelf life of stored fruits, vegetables and tubers. The formulation of this invention is particularly useful for storage of fruits, vegetables and tubers through control of food spoiling microbial species. Hitherto known method of preservation of harvested vegetables consists of either protection through chemical preservatives, toxic pesticides, wax coating or by cold storage. While presence of synthetic chemicals on food products is unacceptable in all countries who are aware of the health hazard caused pesticidal residues on food product. The second technique is almost in accessible to the average farmer. In contrast to this, preservation and extension of the shelf life through the iron chelating surface coating introduced by the present process is a safe and simple process. Through this process it is possible to prevent growth of food micro-organisms through blockage of their iron uptake. By this method it is possible to prevent the deterioration of stored fruits, vegetables and tubers through food spoiling microbial forms without affecting the quality of the produce. The process involves culturing a mutated organism Pseudomonas RRLJ 181 in an iron free conventional nutrient medium as described in the article (Meyer & Abdallah J Gen Microbiol. 107: 319,1978) and induce it to produce a form of Pseudobactin hitherto unknown. This product when sprayed on harvested farm produce such as fruits and vegetables, fonns a protective coating to extend their shelf life without refrigeration. The strain Pseudomonas RRLJ 181 which has been deposited at RRL Jorhat, has the following salient characters RRLJ 181: Yellow slimy round colonies, produces bluishgreen siderophores. Able to grow in a pH range of 5-9. Pigment colour sensitive to pH. It produces bluishgreen pseudobactin in the iron free culture medium of Meyer and Abdallah (J Gen Microbiol. 107: 319,1978). The formulation of the present Invention used for the preservation of harvested vegetables, tubers and fruits is a environment friendly methodology wherein no synthetic chemicals are used. There will be no pesticidal residues on the produce. The process prevents the growth of food spoiling microorganisms by depriving their iron uptake. There will be no toxic effluents produced during the manufacture of this natural preservative and no toxic chemicals are used to produce the formulation. The process involves culturing of the organism by growing the organism under sterile controlled conditions using double distilled water and iron free nutrient medium ( Meyer and Abdallah 1978) in shakers at a pH of 7 in a temperature range of 25-30°C. The product formation is detected visually through the green colour imparted to the medium in 48 h after inoculation of the strain into the culture medium. This is then adsorbed in an XAD column and eluted in methanol. It can also be adsorbed on silica gel and eluted in methanol. This product is diluted to 10:100 concentration and the farm produce to be treated is dipped into this solution for 30 seconds. By employing the formulation, it is possible to prevent the deterioration of stored vegetables and tubers through impediment of bacterial and fungal growth without affecting the quality and safety of the edible marketable products. Mode of action of the formulation of this invention is suppression of the microbial species causing food spoilage through production of iron chelators which deprive the fungal species of the essential element. The antibiotic produced by this organism is an iron chelator and it inhibits bacterial and fungal species attacking the harvested fruits and vegetables by depriving those saprophytes of the essential iron. Thus nature of the antibiotic is completely non toxic and safe from the point of view of environment. Fruits (tomatoes, apples, bananas, papaya) vegetables (string beans, bitter guard) tubers potatoes) were dipped for 30 seconds in the solution containing the psuedobactin preparation produced by the organism and stored without refrigeration when the inside temperature was around 30-35°C and the average humidity was around 90%. The ripe fruits remained intact for 5 days as against 2 days in controls. Vegetables for 7 days as against 3 days in controls. Accordingly the present invention provides an improved process for the preparation of pseudobactin useful for increasing the shelf life of stored fruits , vegetables and tubers , which comprises cultivating a novel strain of Pseudomonus sp. having characteristics such as herein described in a conventional for 48 h iron free medium at pH 7 in a temperature ranging 25 - 30 ° C2 recovering pseudobactin from culture medium by conventional chromatographic methods such as herein described. The growth may be effected for 48h which is sufficient for the organism to excrete into the culture medium copious quantities of the product to be used for preservation of fruits and vegetables. Tests through bioassay are given as illustrations: Example 1: Antibiosis effect against fungal pathogens tested in plating media. Actively growing cells of the strains created a zone of inhibition to the fungai pathogens. Siderophores isolated from cultures grown in iron free medium reduced fungal mycelial growth at lower concentrations and inhibited it completely at higher concentrations. Example 2: Fruits (tomatoes, apples, bananas, papaya), vegetables (string beans, bitter guard) and tubers (potatoes), were dipped for 30 seconds in the culture filtrate of the organism as indicated above and stored without refrigeration when the inside temperature was around 30-35°C and the average humidity was around 90%. The ripe fruits remained intact for 5 days as against 2 days in controls, vegetables for 7 days as against 3 days in controls and tubers for 30 days as against 15 days in controls. Example 3: As a test to confirm fungal inhibition through this treatment, the test fruits, vegetables and tubers were dipped in a conidial suspension of the most common food spoiling fungi Rhizopus nigricans and Fusarium spp. after the treatment with the culture filtrate as described above. The growth of the fungi on the treated produce was markedly reduced. The main properties of this invention are: 1. It protects stored vegetables and fruits in storage and in transport under unrefrigerated conditions. 2. it allows development of efficient and healthier storage practices which does not affect the environment through toxic pollutants. 3. The formulation is a nontoxic product to increase shelflife of stored fruits, vegetables and tubers without effecting the quality and safety of the edible marketable products. We Claim: 1. An improved process for the preparation of pseudobactin useful for increasing the shelf life of stored fruits , vegetables and tubers , which comprises cultivating a novel strain of Pseudomonus sp. having characteristics such as herein described in a conventional for 48h iron free medium at pH 7 in a temperature ranging 25 - 30 ° C2, recovering pseudobactin from culture medium by conventional chromatographic methods such as herein described. 2. An improved process as claimed in claim 1 wherein the recoveiy of pseudobactin is effected by adsorbing in an XAD column and eluting in methanol . 3. An improved process for the preparation of a pseudobactin useful for increasing the shelf life of stored fruits , vegetables and tubers substantially as herein described .

Full Text

This invention relates to an improved process for the preparation of a pseudobactin formulation useful for increasing the shelf life of stored fruits, vegetables and tubers.
The formulation of this invention is particularly useful for storage of fruits, vegetables and tubers through control of food spoiling microbial species.
Hitherto known method of preservation of harvested vegetables consists of either protection through chemical preservatives, toxic pesticides, wax coating or by cold storage. While presence of synthetic chemicals on food products is unacceptable in all countries who are aware of the health hazard caused pesticidal residues on food product. The second technique is almost in accessible to the average farmer.
In contrast to this, preservation and extension of the shelf life through the iron chelating surface coating introduced by the present process is a safe and simple process. Through this process it is possible to prevent growth of food micro-organisms through blockage of their iron uptake. By this method it is possible to prevent the deterioration of stored fruits, vegetables and tubers through food spoiling microbial forms without affecting the quality of the produce.
The process involves culturing a mutated organism Pseudomonas RRLJ 181 in an iron free conventional nutrient medium as described in the article (Meyer & Abdallah J Gen Microbiol. 107: 319,1978) and induce it to produce a
form of Pseudobactin hitherto unknown. This product when sprayed on harvested farm produce such as fruits and vegetables, fonns a protective coating to extend their shelf life without refrigeration.
The strain Pseudomonas RRLJ 181 which has been deposited at RRL Jorhat, has the following salient characters RRLJ 181: Yellow slimy round colonies, produces bluishgreen siderophores.
Able to grow in a pH range of 5-9. Pigment colour sensitive to pH. It produces bluishgreen pseudobactin in the iron free culture medium of Meyer and Abdallah (J Gen Microbiol. 107: 319,1978).
The formulation of the present Invention used for the preservation of harvested vegetables, tubers and fruits is a environment friendly methodology wherein no synthetic chemicals are used. There will be no pesticidal residues on the produce. The process prevents the growth of food spoiling microorganisms by depriving their iron uptake. There will be no toxic effluents produced during the manufacture of this natural preservative and no toxic chemicals are used to produce the formulation.
The process involves culturing of the organism by growing the organism under sterile controlled conditions using double distilled water and iron free nutrient medium ( Meyer and Abdallah 1978) in shakers at a pH of 7 in a temperature range of 25-30°C. The product formation is detected visually through the green colour imparted to the medium in 48 h after inoculation of the strain into the culture medium. This is then adsorbed in an XAD column
and eluted in methanol. It can also be adsorbed on silica gel and eluted in methanol. This product is diluted to 10:100 concentration and the farm produce to be treated is dipped into this solution for 30 seconds.
By employing the formulation, it is possible to prevent the deterioration of stored vegetables and tubers through impediment of bacterial and fungal growth without affecting the quality and safety of the edible marketable products.
Mode of action of the formulation of this invention is suppression of the microbial species causing food spoilage through production of iron chelators which deprive the fungal species of the essential element. The antibiotic produced by this organism is an iron chelator and it inhibits bacterial and fungal species attacking the harvested fruits and vegetables by depriving those saprophytes of the essential iron. Thus nature of the antibiotic is completely non toxic and safe from the point of view of environment.
Fruits (tomatoes, apples, bananas, papaya) vegetables (string beans, bitter guard) tubers potatoes) were dipped for 30 seconds in the solution containing the psuedobactin preparation produced by the organism and stored without refrigeration when the inside temperature was around 30-35°C and the average humidity was around 90%. The ripe fruits remained intact for 5 days as against 2 days in controls. Vegetables for 7 days as against 3 days in controls.
Accordingly the present invention provides an improved process for the
preparation of pseudobactin useful for increasing the shelf life of stored
fruits , vegetables and tubers , which comprises cultivating a novel strain
of Pseudomonus sp. having characteristics such as herein described in a
conventional for 48 h iron free medium at pH 7 in a temperature ranging 25 -
30 ° C2 recovering pseudobactin from culture medium by conventional
chromatographic methods such as herein described.
The growth may be effected for 48h which is sufficient for the organism to excrete into the culture medium copious quantities of the product to be used for preservation of fruits and vegetables.
Tests through bioassay are given as illustrations:
Example 1: Antibiosis effect against fungal pathogens tested in plating media. Actively growing cells of the strains created a zone of inhibition to the fungai pathogens. Siderophores isolated from cultures grown in iron free medium reduced fungal mycelial growth at lower concentrations and inhibited it completely at higher concentrations.
Example 2: Fruits (tomatoes, apples, bananas, papaya), vegetables (string beans, bitter guard) and tubers (potatoes), were dipped for 30 seconds in the culture filtrate of the organism as indicated above and stored without refrigeration when the inside temperature was around 30-35°C and the average humidity was around 90%. The ripe fruits remained intact for 5 days
as against 2 days in controls, vegetables for 7 days as against 3 days in controls and tubers for 30 days as against 15 days in controls.
Example 3: As a test to confirm fungal inhibition through this treatment, the test fruits, vegetables and tubers were dipped in a conidial suspension of the most common food spoiling fungi Rhizopus nigricans and Fusarium spp. after the treatment with the culture filtrate as described above. The growth of the fungi on the treated produce was markedly reduced.
The main properties of this invention are:
1. It protects stored vegetables and fruits in storage and in transport under unrefrigerated conditions.
2. it allows development of efficient and healthier storage practices which does not affect the environment through toxic pollutants.
3. The formulation is a nontoxic product to increase shelflife of stored fruits, vegetables and tubers without effecting the quality and safety of the edible marketable products.

We Claim:
1. An improved process for the preparation of pseudobactin useful for increasing the shelf life of stored fruits , vegetables and tubers , which comprises cultivating a novel strain of Pseudomonus sp. having characteristics such as herein described in a conventional for 48h iron free medium at pH 7 in a temperature ranging 25 - 30 ° C2, recovering pseudobactin from culture medium by conventional chromatographic methods such as herein described.
2. An improved process as claimed in claim 1 wherein the recoveiy of pseudobactin is effected by adsorbing in an XAD column and eluting in methanol .
3. An improved process for the preparation of a pseudobactin useful for increasing the shelf life of stored fruits , vegetables and tubers substantially as herein described .

Documents:

2513-del-1998-abstract.pdf

2513-DEL-1998-Claims.pdf

2513-del-1998-complete specification(granted).pdf

2513-del-1998-correspondence-others.pdf

2513-del-1998-correspondence-po.pdf

2513-del-1998-description (complete).pdf

2513-del-1998-form-1.pdf

2513-del-1998-form-2.pdf

2513-del-1998-form-3.pdf

2513-del-1998-form-9.pdf

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Patent Number

189747

Indian Patent Application Number

2513/DEL/1998

PG Journal Number

16/2003

Publication Date

19-Apr-2003

Grant Date

27-Jan-2004

Date of Filing

26-Aug-1998

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH,

Applicant Address

RAFI MARG NEW DELHI-110001, INDIA

Inventors:

#	Inventor's Name	Inventor's Address
1	BALAMONI BEZBARUAH	REGIONAL RESEARCH LABORATORY,JORHAT,ASSAM,INDIA
2	DILEEP KUMAR BHASKANANNAIR SARASWATHYAMMA	REGIONAL RESEARCH LABORATORY,JORHAT,ASSAM,INDIA

PCT International Classification Number

A01F 25/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA