Title of Invention	"A PROCESS FOR THE PREPARATION OF AN IMPROVED SYNERGISTIC COMPOSITION USEFUL AS GROWTH MEDIUM FOR FUNGI AND BACTERIA"
Abstract	A process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which comprises dissolving the carbon nitrogen and trace element sources in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml, getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as i. Beef extract 8-12 gm, ii. Starch 1-2 gm, iii. Dextrose 10-12 gm, iv. Proteose peptone 1-3 gm, v. Tryptose 1-3 gm, vi. Yeast extract 1-3 gm, vii. Coblat chloride or nitrate 5-10mg viii. Ferric ammonium citrate 5-1 Omg ix. Magnesium sulphate or chloride 0-10mM x. Calcium chloride or nitrate 0-10mM,

Title of Invention

"A PROCESS FOR THE PREPARATION OF AN IMPROVED SYNERGISTIC COMPOSITION USEFUL AS GROWTH MEDIUM FOR FUNGI AND BACTERIA"

Abstract

A process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which comprises dissolving the carbon nitrogen and trace element sources in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml, getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as i. Beef extract 8-12 gm, ii. Starch 1-2 gm, iii. Dextrose 10-12 gm, iv. Proteose peptone 1-3 gm, v. Tryptose 1-3 gm, vi. Yeast extract 1-3 gm, vii. Coblat chloride or nitrate 5-10mg viii. Ferric ammonium citrate 5-1 Omg ix. Magnesium sulphate or chloride 0-10mM x. Calcium chloride or nitrate 0-10mM,

Full Text	This invention relates to the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria. The medium is found to be efficient growth promoter particularly for vegetative growth of large number of microorganisms which are otherwise known to be fastideous. Examples of such organisms are: species of the genera of Actinomadura, Amycolata, Arthrobacter, Brevibacterium, Corynebacterium, Rhodococcus, Streptomyces, Streptoverticillium, Saccharomonospora, Saccharopolyspora, Streptosporangium, Streptococcus, Staphylococcus, Microbispora, Microtetraspora, Micromonospora, Nocardia, Nocardiodes, slow and fast growing Mycobacteria, Bacillus, Eschericia, Pseudomonas as well as several slow growing but yet unidentified actinomycetes and several groups of fungi. Several known media such as (1) Luria Bertani (LB) having the composition per 1000ml : lOgm Tryptone, 5gm Yeast extract, lOgm Sodium Chloride; (2) Yeast extract : malt extract per 1000ml: 2gm Yeast extract, 4gm Malt extract, lOgm Glucose; (3) Tryptic Soya broth per 1000ml : 17gm Pancreatic Digest of casein, 3gm Papaic digest of Soya meal, 5gm Sodium Chloride, 2.5gm Dibasic Potassium Phosphate, 2.5gm Dextrose and (4) Bennett's medium containing per 1000ml : Igm yeast extract, Igm Beef Extract, 2gm Pancreatic digest of casein, lOgm Dextrose, were tested and found that none of them were satisfactory growth promotor for the strains and groups listed above. In the case of Bennett's medium, organisms like Actinomadura, Streptomyces, Nocardia, Streptosporangium, Microbispora, Microtetraspora, Micromonospora can take 10-15 days or even longer to show full growth (late exponential phase) however genera such as Mycobacterium, Streptococcus and Staphylococcus do not grow at all . Likewise, most of the mycobacteria are very fastideous and require special media components and supplementation of trace elements, vitamins, fatty acids etc. Further, for several experiments early sporulation is an undesirable property which is encountered many a times while using media such as LB, Bennett's and Yeast extract: malt extract. Therefore, whenever large mycelial growth (Vegetative growth) is required almost all the media mentioned above are not very useful. The medium described in the present invention is useful for the growth of wide group of organisms such as actinomycetes, several other Gram+ bacteria, Gram- bacteria and several groups of fungi. The medium of present invention particularly helps large mycelial growth and delays sporulation. The medium of present invention also reduces the time required for growth more particularly of the group actinomycetes. The present invention is thus based upon the observation that the known media listed as above are not easily assimilable therefore not very useful for desired growth. Further, all the media lacked trace elements which are very important for the growth of live organisms. Thus the main objective of this invention is to prepare a universal growth medium which supports the growth of a large number of organisms belonging to various groups such as fungi, Gram- and Gram+ bacteria but more particular of Actinomycetes. Other objectives of this invention is to develop a medium which can shorten the time required for growth, which can produce large amount of exponentially growing cells and also which can withstand relatively high temperature of growth without having an adverse effect on the growth pattern. Thus, this invention provides for the preparation of a universal growth medium useful for fungi and bacteria. One of the most important aspect of the present invention is the addition of extra carbon source to the medium. It is well known that glucose enters the metabolic pathway without any modification thus it is a preferred substrate of carbon for most of the organisms and gets utilized very quickly. However, high concentration of glucose in the medium can have adverse effect because glucose changes the osmolarity of the medium. Glucose/dextrose concentration in the medium is also influenced by autoclaving and long incubation period. Many of the actinomycetes genera are relatively slow growing thus there is a need to provide an additional carbon source which can release glucose/dextrose slowly and steadly for longer period. Looking at the properties of actinomycetes and some fungi, starch was chosen as an additional source of carbon because most of the organisms listed above can utilize starch, albeit slowly. Another important aspect of the present invention is the supplementation of trace elements such as cobalt and iron. Crucial role played by trace elements in the growth of a organism is a well established fact. However, none of the media listed above have trace elements as part of their component. Thus, a new growth medium was prepared by mixing the appropriate quantities of the following chemicals and complex media components such as : Beef extract or Lab Lamco, Yeast extract, Tryptose, Proteose peptone, Soluble starch, Dextrose, and traces of Cobalt chloride or Cobalt nitrate and Ferric ammonium citrate. The composition when established and dissolved in double glass- distilled water, at room temperature, has a pH from about 6.0 to 6.6. This medium when used as it is, or after adjusting the pH between 7.0 and 7.2 by using IN Sodium hydroxide acts as a universal medium for the growth of large number of actinomycetes genera, other Gram+ bacteria, some Gram- bacteria and for several groups of fungi. Accordingly the present invention provides a process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which comprises dissolving the carbon ,nitrogen and trace element source in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml, getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as i. Beef extract 8-12 gm, ii. Starch 1-2 gm, iii. Dextrose 10-12 gm , iv. Proteose peptone 1 -3 gm, v tryptose 1-3 gm, vi yeast extract l-3gm, vii. Cobalt chloride or nitrate 5-10 mg viii Ferric ammonium citrate 5-10 mg ix Magnesium sulphate or chloride 0-10 mM x. Calcium chloride or nitrate 0-10 mM , The main embodiment of the present medium may contain macronutrients such as carbon in the form of Lab Lamco or Beef extract, starch and dextrose, nitrogen in the form of proteose peptone and tryptose, micronutrients such as vitamins in the form of yeast extract, trace elements such as cobalt chloride or cobalt nitrate and ferric ammonium citrate and double glass- distilled water. Purified agar, calcium and magnesium may be added optionally when required. The concentration of the different macro and micro nutrients used in the medium of the present invention are as follows: the macronutrient are: Lab lamco 8-10 gm or Beef Extract 10-12 gm, proteose peptone 1-3 gm , tryptose 1-3 gm, dextrose 10-12 gm, soluble starch 1-2 gm, and the micronutrients are Yeast extract 1-3 g and trace elements are Cobalt chloride or nitrate 5-10 mg, ferric ammonium citrate 5-10 mg, magnesium sulphate or magnesium chloride 5-10mM, calcium chloride or calcium nitrate 5-10mM, double glass distilled water 1000 ml, pH 6.8 to 7.2. The addition of calcium and magnesium ions is optional, its presence or absence does not particularly affect the growth of every organism. However the preferred concentration of each of the components listed above is as detailed in the example number 1. The process of the present invention is illustrated through the examples given below, which should not however be construed to limit the scope of the present invention. Example 1: Following media components and analytical grade chemicals were mixed at room temperature: Beef extract lOgm, yeast extract 2gm, proteose peptone 2 gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass- distilled water 1000ml, Agar 1 Igm, pH 7.0. The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. This medium supported the growth of Actinomadura sps, Amycolata sps, Arthrobacter sps, Brevibacterium sps, Corynebacterium sps, Rhodococcus sps, large number of Streptomyces sps, Saccharomonospora sps, Saccharopolyspora sps, Microbispora sps, Microtetraspora sps, Micromonospora sps, Nocardia sps, Nocardiodees sps, and Streptosporangium sps, Streptococcus sps, Staphylococcus sps, Bacillus sps, Eschericia sps, Pseudomonas sps and several groups of fungi. Species of mycobacteria did not grow very well in this medium. Poor sporulation of some species of Streptomyces, Microbispora, Microtetraspora was observed after 15 days of growth, however most of the strains listed above did not sporulate at all and produced good mycelial growth after 3 days of incubation. Thus establishing that the medium will be very useful for all the actinomycetes and other groups of bacteria and fungi. The species of Saccharomonospora grew well in this medium upto 55°C. Example 2: The medium composition was Beef extract 12 gm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass distilled water 1000ml, Agar llgm, pH 7.0. The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. Slow growing species of Actinomadura, Streptosporangium, Microbispora and Microtetraspora grew poorly as compared to the medium composition which was supplemented with lOgm/1000ml beef extract and starch as described above. Example 3: The medium composition was Beef extract lOgm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass distilled water 1000ml, Agar llgm, pH 7.0 . The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. Keeping rest of the media component same, as described above, dextrose was replaced with 1% v/v glycerol as final concentration and 1.0 gm citric acid was added. These replacement and addition facilitated the growth of wide range of fast growing mycobacteria and atleast one slow growing mycobacterium that isM.bovis BCG (Bacillus Calmette Guerin). Example 4 : The medium composition was Beef extract lOgm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, 1% v/v glycerol, Igm citric acid, double glass- distilled water 1000ml, Agar llgm, pH 7.0. The medium was sterilized at 20Lb per inch sqare (steam) pressure for 20 minutes. Growth of the Mycobacteria sps were still better in this embodiment than in the example No. 3. Good growth was observed after 5 days of incubation at 37°C. This result was true for the slow growing Mycobacterium bovis BCG also. Example 5: The medium composition was Beef extract 12gm, yeast extract 2gm, peptone 2gm, tryptone 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass- distilled water 1000ml, Agar llgm, pH 7.0 . The medium was sterilized at 20 Lb per inch square (steam) pressure for 20 minutes. These changes were not very satisfactory as all the strains showed relatively slow growth. However, this example also gave better results than LB, yeast extract: malt extract and Bennett's medium. Example 6: The medium containing Beef extract lOgm, yeast extract 2gm, Proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, Magnesium sulfate l0mM (mili molar) Calcium nitrate 10mM(mili molar), agar 11gm, double glass- distilled water 1000ml pH 7.0 was inoculated with species of the genus Saccharomonospora and inubated at 55°C. The organism showed good growth in the medium described above. The examples described clearly establish that the present invention is an improvement over the existing media used for the growth of large number of micro-organisms especially actinomycetes. The medium also supports the growth of several groups of fungi including the fastideous group basidiomycetes. The medium shortens the growth time required and as the medium does not support sporulation, it is useful whenever large mycelial culture is required. The above examples establish that the medium composition described in this invention is a synergistic composition having different aggregate properties then properties of individual components and due to this syngergism the growth medium helps in the faster growth of wide range of organisms, does not allow sporulation and can withstand relatively high growth temperature without affecting the growth pattern and after some modifications as explained in the examples 3 and 4, even the fastideous organisms such as mycobacteria can easily be grown. The examples described clearly establish that the present invention is an improvement over the existing media used for the growth of large number of micro-organisms especially actinomycetes. The medium also supports the growth of several groups of fungi including the fastideous group basidiomycetes. The medium shortens the growth time required and as the medium does not support sporulation, it is useful whenever large mycelial culture is required. Claims: We claim: 1. A process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which dissolving the carbon ,nitrogen and trace element sources in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml , getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as 1. Beef extract 8-12 gm, ii. Starch 1-2 gm, iii. Dextrose 10-12 gm , iv. Proteose peptone 1-3 gm, v tryptose 1-3 gm, vi yeast extract l-3gm, vii. Cobalt chloride or nitrate 5-10 mg viii Ferric ammonium citrate 5-10 mg ix Magnesium sulphate or chloride 0-10 mM xi. Calcium chloride or nitrate 0-10 mM , 2. A process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria substantially as herein described with reference to the Examples.

Full Text

This invention relates to the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria. The medium is found to be efficient growth promoter particularly for vegetative growth of large number of microorganisms which are otherwise known to be fastideous. Examples of such organisms are: species of the genera of Actinomadura, Amycolata, Arthrobacter, Brevibacterium, Corynebacterium, Rhodococcus, Streptomyces, Streptoverticillium, Saccharomonospora, Saccharopolyspora, Streptosporangium, Streptococcus, Staphylococcus, Microbispora, Microtetraspora, Micromonospora, Nocardia, Nocardiodes, slow and fast growing Mycobacteria, Bacillus, Eschericia, Pseudomonas as well as several slow growing but yet unidentified actinomycetes and several groups of fungi.
Several known media such as (1) Luria Bertani (LB) having the composition per 1000ml :
lOgm Tryptone, 5gm Yeast extract, lOgm Sodium Chloride; (2) Yeast extract : malt extract per
1000ml: 2gm Yeast extract, 4gm Malt extract, lOgm Glucose; (3) Tryptic Soya broth per 1000ml
: 17gm Pancreatic Digest of casein, 3gm Papaic digest of Soya meal, 5gm Sodium Chloride, 2.5gm
Dibasic Potassium Phosphate, 2.5gm Dextrose and (4) Bennett's medium containing per
1000ml : Igm yeast extract, Igm Beef Extract, 2gm Pancreatic digest of casein, lOgm Dextrose, were tested and found that none of them were satisfactory growth promotor for the strains and groups listed above. In the case of Bennett's medium, organisms like Actinomadura, Streptomyces, Nocardia, Streptosporangium, Microbispora, Microtetraspora, Micromonospora can take 10-15 days or even longer to show full growth (late exponential phase) however genera such as Mycobacterium, Streptococcus and Staphylococcus do not grow at all . Likewise, most of the mycobacteria are very fastideous and require special media components and supplementation of trace elements, vitamins, fatty acids etc. Further, for several experiments early sporulation is an undesirable property which is encountered many a times while using media such as LB, Bennett's and Yeast extract: malt extract. Therefore, whenever large mycelial growth (Vegetative growth) is required almost all the media mentioned above are not very useful.
The medium described in the present invention is useful for the growth of wide group of organisms such as actinomycetes, several other Gram+ bacteria, Gram- bacteria and several groups of fungi. The medium of present invention particularly helps large mycelial growth and delays sporulation. The medium of present invention also reduces the time required for growth more particularly of the group actinomycetes.

The present invention is thus based upon the observation that the known media listed as above are not easily assimilable therefore not very useful for desired growth. Further, all the media lacked trace elements which are very important for the growth of live organisms.
Thus the main objective of this invention is to prepare a universal growth medium which supports the growth of a large number of organisms belonging to various groups such as fungi, Gram- and Gram+ bacteria but more particular of Actinomycetes. Other objectives of this invention is to develop a medium which can shorten the time required for growth, which can produce large amount of exponentially growing cells and also which can withstand relatively high temperature of growth without having an adverse effect on the growth pattern.
Thus, this invention provides for the preparation of a universal growth medium useful for fungi and bacteria.
One of the most important aspect of the present invention is the addition of extra carbon source to the medium. It is well known that glucose enters the metabolic pathway without any modification thus it is a preferred substrate of carbon for most of the organisms and gets utilized very quickly. However, high concentration of glucose in the medium can have adverse effect because glucose changes the osmolarity of the medium. Glucose/dextrose concentration in the medium is also influenced by autoclaving and long incubation period. Many of the actinomycetes genera are relatively slow growing thus there is a need to provide an additional carbon source which can release glucose/dextrose slowly and steadly for longer period. Looking at the properties of actinomycetes and some fungi, starch was chosen as an additional source of carbon because most of the organisms listed above can utilize starch, albeit slowly. Another important aspect of the present invention is the supplementation of trace elements such as cobalt and iron. Crucial role played by trace elements in the growth of a organism is a well established fact. However, none of the media listed above have trace elements as part of their component.
Thus, a new growth medium was prepared by mixing the appropriate quantities of the following chemicals and complex media components such as : Beef extract or Lab Lamco, Yeast extract, Tryptose, Proteose peptone, Soluble starch, Dextrose, and traces of Cobalt chloride or Cobalt nitrate and Ferric ammonium citrate. The composition when established and dissolved in double glass- distilled water, at room temperature, has a pH from about 6.0 to 6.6. This medium when used as it is, or after adjusting the pH between 7.0 and 7.2 by using IN Sodium hydroxide acts as a universal medium for the growth of large number of actinomycetes genera, other Gram+ bacteria, some Gram- bacteria and for several groups of fungi.

Accordingly the present invention provides a process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which comprises dissolving the carbon ,nitrogen and trace element source in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml, getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as
i. Beef extract 8-12 gm,
ii. Starch 1-2 gm,
iii. Dextrose 10-12 gm ,
iv. Proteose peptone 1 -3 gm, v tryptose 1-3 gm,
vi yeast extract l-3gm,
vii. Cobalt chloride or nitrate 5-10 mg
viii Ferric ammonium citrate 5-10 mg
ix Magnesium sulphate or chloride 0-10 mM
x. Calcium chloride or nitrate 0-10 mM ,
The main embodiment of the present medium may contain macronutrients such as carbon in the form of Lab Lamco or Beef extract, starch and dextrose, nitrogen in the form of proteose peptone and tryptose, micronutrients such as vitamins in the form of yeast extract, trace elements such as cobalt chloride or cobalt nitrate and ferric ammonium citrate and double glass- distilled water. Purified agar, calcium and magnesium may be added optionally when required.
The concentration of the different macro and micro nutrients used in the
medium of the present invention are as follows: the
macronutrient are: Lab lamco 8-10 gm or Beef Extract 10-12 gm,
proteose peptone 1-3 gm , tryptose 1-3 gm, dextrose 10-12 gm, soluble starch 1-2 gm, and the micronutrients are Yeast extract 1-3 g and trace elements are Cobalt

chloride or nitrate 5-10 mg, ferric ammonium citrate 5-10 mg, magnesium sulphate or magnesium chloride 5-10mM, calcium chloride or calcium nitrate 5-10mM, double glass distilled water 1000 ml, pH 6.8 to 7.2. The addition of calcium and magnesium ions is optional, its presence or absence does not particularly affect the growth of every organism. However the preferred concentration of each of the components listed above is as detailed in the example number 1.
The process of the present invention is illustrated through the examples given below, which should not however be construed to limit the scope of the present invention.
Example 1: Following media components and analytical grade chemicals were mixed at room temperature: Beef extract lOgm, yeast extract 2gm, proteose peptone 2 gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass- distilled water 1000ml, Agar 1 Igm, pH 7.0. The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. This medium supported the growth of Actinomadura sps, Amycolata sps, Arthrobacter sps, Brevibacterium sps, Corynebacterium sps, Rhodococcus sps, large number of Streptomyces sps, Saccharomonospora sps, Saccharopolyspora sps, Microbispora sps, Microtetraspora sps, Micromonospora sps, Nocardia sps, Nocardiodees sps, and Streptosporangium sps, Streptococcus sps, Staphylococcus sps, Bacillus sps, Eschericia sps, Pseudomonas sps and several groups of fungi. Species of mycobacteria did not grow very well in this medium. Poor sporulation of some species of Streptomyces, Microbispora, Microtetraspora was observed after 15 days of growth, however most of the strains listed above did not sporulate at all and produced good mycelial growth after 3 days of incubation. Thus establishing that the medium will be very useful for all the actinomycetes and other groups of bacteria and fungi. The species of Saccharomonospora grew well in this medium upto 55°C.
Example 2: The medium composition was Beef extract 12 gm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass distilled water 1000ml, Agar llgm, pH 7.0. The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. Slow growing species of Actinomadura, Streptosporangium, Microbispora and Microtetraspora grew poorly as compared to the medium composition which was supplemented with lOgm/1000ml beef extract and starch as described above.

Example 3: The medium composition was Beef extract lOgm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass distilled water 1000ml, Agar llgm, pH 7.0 . The medium was sterilized at 20Lb per inch square (steam) pressure for 20 minutes. Keeping rest of the media component same, as described above, dextrose was replaced with 1% v/v glycerol as final concentration and 1.0 gm citric acid was added. These replacement and addition facilitated the growth of wide range of fast growing mycobacteria and atleast one slow growing mycobacterium that isM.bovis BCG (Bacillus Calmette Guerin).
Example 4 : The medium composition was Beef extract lOgm, yeast extract 2gm, proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, 1% v/v glycerol, Igm citric acid, double glass- distilled water 1000ml, Agar llgm, pH 7.0. The medium was sterilized at 20Lb per inch sqare (steam) pressure for 20 minutes. Growth of the Mycobacteria sps were still better in this embodiment than in the example No. 3. Good growth was observed after 5 days of incubation at 37°C. This result was true for the slow growing Mycobacterium bovis BCG also.
Example 5: The medium composition was Beef extract 12gm, yeast extract 2gm, peptone 2gm, tryptone 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, double glass- distilled water 1000ml, Agar llgm, pH 7.0 . The medium was sterilized at 20 Lb per inch square (steam) pressure for 20 minutes. These changes were not very satisfactory as all the strains showed relatively slow growth. However, this example also gave better results than LB, yeast extract: malt extract and Bennett's medium.
Example 6: The medium containing Beef extract lOgm, yeast extract 2gm, Proteose peptone 2gm, tryptose 2gm, dextrose lOgm, Soluble starch Igm, Cobalt chloride 5mg, ferric ammonium citrate 5mg, Magnesium sulfate l0mM (mili molar) Calcium nitrate 10mM(mili molar), agar 11gm, double glass- distilled water 1000ml pH 7.0 was inoculated with species of the genus Saccharomonospora and inubated at 55°C. The organism showed good growth in the medium described above.
The examples described clearly establish that the present invention is an improvement over the existing media used for the growth of large number of micro-organisms especially actinomycetes. The medium also supports the growth of several groups of fungi including the fastideous group basidiomycetes. The medium shortens the growth time required and as the medium does not support sporulation, it is useful whenever large mycelial culture is required.

The above examples establish that the medium composition described in this invention is a synergistic composition having different aggregate properties then properties of individual components and due to this syngergism the growth medium helps in the faster growth of wide range of organisms, does not allow sporulation and can withstand relatively high growth temperature without affecting the growth pattern and after some modifications as explained in the examples 3 and 4, even the fastideous organisms such as mycobacteria can easily be grown.
The examples described clearly establish that the present invention is an improvement over the existing media used for the growth of large number of micro-organisms especially actinomycetes. The medium also supports the growth of several groups of fungi including the fastideous group basidiomycetes. The medium shortens the growth time required and as the medium does not support sporulation, it is useful whenever large mycelial culture is required.

Claims:
We claim:
1. A process for the preparation of an improved synergistic composition useful as growth medium for fungi and bacteria which dissolving the carbon ,nitrogen and trace element sources in double distilled water and mixing the dissolved ingredients by conventional manner to make the volume to 1000 ml , getting the desired synergistic composition wherein the carbon, nitrogen and trace element sources are such as
1. Beef extract 8-12 gm,
ii. Starch 1-2 gm,
iii. Dextrose 10-12 gm ,
iv. Proteose peptone 1-3 gm,
v tryptose 1-3 gm,
vi yeast extract l-3gm,
vii. Cobalt chloride or nitrate 5-10 mg
viii Ferric ammonium citrate 5-10 mg
ix Magnesium sulphate or chloride 0-10 mM
xi. Calcium chloride or nitrate 0-10 mM ,
2. A process for the preparation of an improved synergistic composition useful as
growth medium for fungi and bacteria substantially as herein described with
reference to the Examples.

Documents:

753-del-1998-abstract.pdf

753-del-1998-claims.pdf

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

753-del-1998-correspondence-others.pdf

753-del-1998-correspondence-po.pdf

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

753-del-1998-form-1.pdf

753-del-1998-form-2.pdf

753-del-1998-form-4.pdf

« Previous Patent

Next Patent »

Patent Number

188337

Indian Patent Application Number

753/DEL/1998

PG Journal Number

36/2002

Publication Date

07-Sep-2002

Grant Date

06-Jun-2003

Date of Filing

24-Mar-1998

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRAIL RESEARCH

Applicant Address

RAFI MARG, NEW DEHI-110 001, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	DR. PUSHPA AGRAWAL	MICROBIAL TECHNOLOGY, SECTOR. 39-A, CHANDIGARGH- 160 036 (INDIA)

PCT International Classification Number

A01H 005/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA