Title of Invention	"AN IMPROVED PROCESS FOR THE PRODUCTION OF 4,6,6, TRIMETHYL BICYCLO (3.1.1) HEPT-3EN-2ONE USING A NOVEL PENICILLIUM SP."
Abstract	An improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in described in a potato dextrose broth medium supplemented with six carbon sugar, assimilable phosphate yeast extract and alpha pinene at temperature ranging 25-30°C for 6h and recovering the 4,6,6, triethyl bicyclo (3.1.1) hept 3-en-2-one by known methods such as here in described.

Title of Invention

"AN IMPROVED PROCESS FOR THE PRODUCTION OF 4,6,6, TRIMETHYL BICYCLO (3.1.1) HEPT-3EN-2ONE USING A NOVEL PENICILLIUM SP."

Abstract

An improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in described in a potato dextrose broth medium supplemented with six carbon sugar, assimilable phosphate yeast extract and alpha pinene at temperature ranging 25-30°C for 6h and recovering the 4,6,6, triethyl bicyclo (3.1.1) hept 3-en-2-one by known methods such as here in described.

Full Text	The present invention relates to an improved process for the production of the 4,6,6, trimethyl bicyclo (3.1.1) hept-3en-2one using a novel Penicillium sp. More particularly the present invention relates to a process for production of 4,6,6, trimethyl bicyclo (3.1.1) hept - 3en - 2one having formula 1 as shown in accompanying drawing by biotransformation using a novel Penicillium sp. Production of food flavours employing microbi al processes is attractive from the view point of process economy in the utilisation of inexpensive and readily available precursor compounds. These are valued as flavour chemicals and are utilised as food additives. The conversion assumes commercial importance because the starting materials are low-cost and yield value added products. The production of 4,6,6, trimethyl bicyclo [3.1.1]hept 3-en-2one (fig 1), which is the subject of present study comes from the oil of verbena, dill, spearmint and eucalyptus oil (Guenther.E., The Essential Oils, (1952) vol 2.444. Fritzche Brothers, Inc. New York, Publisher: D.Van Nostrand Company Inc. It is commonly known as verbenone with structural formula as shown in the drawing accompanying this specification. Leaves, the natural source, are mainly sold for the herb and drug trade, and small quantities are used for distillation of the essential oil. The distillation process gives a very small yield of the oil generally varying from 0.072% to 0.19% oil. By the present method it can be produced at economical price and would be a valuable adjunct to food and can be used to great advantage in certain types of flavours. The current market price of 4,6,6, trimethyl bicyclo [3.1.1] hept-3en-2one is about Rs.80,000 per kg. It can be formed through microbial source, via hydrogenated bicyclic monoterpene hydrocarbon as shown by Ramadevi, J. and Bhattacharyya. P.K. (1978). Molecular rearrangements in the microbiological transformations of terpenes and chemical logic of microbial process. J. Ind. Chem. Soc, Vol.IV, 1131-1137. The monoterpene alpha-pinene is a by product in lime peel oil, is mainly a waste product from food processing industries. It is also available from turpentine oil as a by product. The utilisation of the low cost substrate to 4,6,6, trimethyi bicycio [3.1.1] hept-3en-2one, a nign value and high priced flavour compound is utilised by a bakeries, confectionary, ice cream and soft drink industry. The compound itself does not have any nutritional value, however, it increases the flavour of the food items. Recently, this subject has received a lot of attention. Although work is being done abroad on similar lines of biotransformation (Nakayama. H, 1978; Cadwallader, 1989; Cheetham, 1995) Nakayama. H, Yamaguchi. Y, Machida, Iwasaki.S, Komtsu.A and Shinoda.A D-L-Menthol (1978) Japan. Kokai Source: CA , 8 9 , 7 4217K. Cadwallader.K.R, Braddock.R.J, Parish.M.E and Higgins.D.P Bioconversion of (+) limonene by Pseudomo-nas gladioli, (1989) Journal of Food Science vol 54 1241-45. Cheetham Peter.S.J. Biotransformations; New Routes to food ingredients. (1995) Chemistry and Industry 3 April. There is no patent or process developed for biotransformation of this particular compound using Penicillium sp. There is no work of similar kind being carried out. Therefore, there is an urgent need to have this type of a process which will help in the utilisation of monoterpene hydrocarbon alpha pinene, a by propduct from the waste of the food industries. A few (four) patents exist on the production of verbenol, 4,6,6-trimethyl bicyclo [3.1.1]hept-2-en-2-ol utilising yeast and bacterial cultures. The patent (No. Int. Pat. Class: C07c-035/28;C 12N-001/26:C12P- 007/02.) by Repph et al show the microbial oxidation from sugar acids to 4, 6, 6 -trimethy1 bicyclo[3.1.1]hept-2-en-2-ol verbenol, but not to bicyclo- [3.1.1] hept-3en-2-one , verbenone utilising Acetobacter, Gluconobacter, Pseudomonas, Aspergillus and Penicillium. In his patent, Repph et al. has mentioned the formation of 4,6,6, trimethyl bicyclo [3.1.1] hept-3en-2-one along with major component 4,6,6- trimethyl bicyclo [3.1.1] hept- 2-en-2-ol utilising methylotrophi Hansenula polvmorpha strain. There is no patent till date for the production of 4,6,6, trimethyl bicyclo- [3.1.1] hept- 3-en-2one from bicyclic monoterpene hydrocarbon utilizing Penicillium Sp. The main objective of the invention is to produce verbenone a value flavour compound which has wide usage in food industries and used as a food flavour additive starting from alpha pinene a by product from lime peel oil which is a waste from food industries. The other object of the present invention is to produce verbenone using a novel strain of Penicillium. Another object is to attain high yeild . Still another object is to supplement the nutrient medium with special components to obtain a high yield. Accordingly the present invention provides an improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in described in a potato dextrose broth medium supplemented with six carbon sugar, assimilable phosphate yeast extract and alpha pinene at temperature ranging 25- 30°C for 6h and recovering the 4,6,6, trimethyl bicyclo (3.1.1) hept 3- en-2-one by known methods such as here in described. In the persent investigation the compound 4,6,6, trimethyl bicyclo (3.1.1) hept -3-en-2-one has been produced starting from bicyclic monoterpene hydrocarbon. The production for the maximum yield has been optimised and media composition has also been varied to obtain the desired results. Concentrations of carbon, yeast extract and phosphorus were shown to affect the yield to a large extent. Microbial transformation specially for the production of food flavours is a recent technique. Earlier some workers have tried biotransformation studies using the substrate alpha pinene as sole source of carbon but in the present study the substrate was varied over a range and it was found that after certain limit the substrate is toxic to the cells and therefore it has been selected by its capacity for the conversion under appropriate growth conditions. The organism was isolated from the soil collected from different areas. The organism isolated from the soil under the lime tree had a higher potency i.e. capacity for bioconversion, therefore, the organism were selected and isolated from the soil under the lime tree where expectedly the microorganisms had adapted themselves to grow on monoterpene hydrocarbons. The underlining principle in the present invention is the use of Surface Response Method for obtaining the maximum conversion yield of 15-2 0%. In the present invention it has been possible to reduce the time of conversion (6 h) , where -as earlier studies indicate a much as 40 to 44 hours. Ramadevi.J and Bhattacharyya.P.K Microbiological transformations of terpenes: part XXIII fermentation of geraniol, nerol and limonene by a soil pseudomonad, Pseudomonas incognita (linalool strain) Ind. J. Biochem. and Biophys. 14, 1977, 288-91. Similarly the yield of the flavouring compounds reported in the patents on conversion of bicyclic monoterpene hydrocarbn to bicyclo- [3.1.1] hept-3-en-2-one has been (10%) reported in the patents on the conversion of bicyclo monoteropene hydrocarbon only by an yeast isolate, Hansenula Repph.H.D Stottme-ist.V ,Duresch.R, Richter.H.P, Schmidt.J, Dorr.M, Hanfe.G, Weber.L (1988) Int. patent. At present there is no process available as mentioned here. It is, therefore, felt that the present data is of great importance or advantage for the food industries. The novelty of the invention lies in use of Peni-cillium sp. for biotransformation of alpha pinene to 4,6,6, trimethyl bicylo (3.1.1) hept 3en-2one and maxi mising the yield much more than which is produced by the plant sources or by any other natural source being used in hither to known processes. Today, there is a lot of awareness especially for food additives like the use of natural flavours and colorants as, more and more people are going in for natural flavours. To produce flavours through microbial source is also natural process. The invention is related to a use of novel strain of organism being used to produce a product,by a bio-technological process. The culture has been deposited in CFTRI culture collection centre, and designated as CFR 201203 dated 10-4-96. For the bioconversion, the culture was isolated from the soil. This was subsequently serially diluted and plated on potato dextrose agar medium. The plates were incubated at 25° to 35°C for a period of 3-10 days. The spores were taken for the experiment. The isolated strain of penicillium sp has the following characteristics the spores are greyish in colour, grows well on potato dextrose agar at 3 0°C the spores have circular margins. The pigment produced slightly diffuses into the agar medium. Hence the present invention provides a biotechno-logical process mediated by a selected strain of Peni-cillium for the production of 4,6,6, trimethyl bicy-clo- [3.1.1]hept-3-en-2- one, generally described as below: A process for the production of 4,6,6, trimethyl bicyclo- [3.1.1]hept-3-en-2one was obtained by modifying the medium and supplementing it with additional six carbon sugar, phosphate and yeast extract. The process comprises of the following steps. 1. The culture is maintained on Potato dextrose agar at 30°C 2. Greyish spores were taken in potato dextrose broth at 3 0°C and grown for 16.5 h. 3. The grown cells were incubated with substarate alpha pinene for 6 h at 120 rpm at 30°C in phosphate buffer at pH 7.0. 4 . The product was extracted in dichloromethane and analyzed on GC. The invention is further illustrated by the following examples, which should not be construed to limit the scope of the invention. Example-1 The spore suspension was taken in potato dextrose broth with an initial cell density 3.0 O.D on spectrophotometer. This was incubated at 30°C, 120 rpm and taken in buffer at pH 7.0 With addition of substrate alpha pinene. The percent yield conversion was 3.2% or 640 ug/ 100ml to the desired product. The product was confirmed on gas chromatography along with the standard and also by gas chromatography mass spectrophotometer fragmentation pattern. GC analysis was done on a Shimadzu Gas chromatograph equipped with a stainless steel SE-30 column and FID detector. For GCMS (gas chromatography-mass spectrophotometry) data, gas Chromatograph mass spectrophotometer Hewlett-Packard 5995 was used with an HP-20 M (carbowax 20M) column (25m x0.32mm) temperature programme at 60°C (6)-4°C/min to 250°C, C15°C, the gas flow rate: lml/min). 4,6,6, trimethyl bicyclo [3.1.1]hept 3-en-2one content was calculated on the area obtained on gas chromatograph of the samples against the area of the standard compound and characterised by GCMS fragmentation pattern. Example-2 The spore suspension with initial cell density of 3.0 O.D was taken in potato dextrose medium supplemented with glucose 7% and after incubation at 30°C was taken up in buffer at pH 7.0 with the substrate. The percent yield conversion increased with such cells to 3.36% or 672 ug/100 ml. The increase in glucose concentration supports the product formation. Example-3 The above spore suspension with additional glucose in the broth was also supplemented with additional inorganic phosphate KH2P04(500mg/l) and yeast extract 250mg/l.The cell pellet was taken up in phosphate buffer with pH 7.0 incubated with substrate alpha pinene for 6 h. This resulted in increasing the yield conversion from 15 to 20% (3000 -4000ug/ 100ml) . Additional glucose, KH2P04 and also yeast extract enhanced the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en- 2-one . The present invention provides a process for the production of 4,6,6, trimethyl bicyclo - [3.1.1] hept-3-en-2-one with number of advantages. 1. A fungal culture that utilises a monoterpene hydro carbon isolated from the waste product of essential oil industry has been isolated. 2. This organism is capable of converting low cost essential oil constituent of citrons oil in to a high valued flavouring compound that has demand in the food processing industry. 3. There is no fungal culture known to our knowledge till date producing bicyclo - {3.1.1} hept - 3 - en - 2 - one. We Claim: 1. An improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in described in a potato dextrose broth medium supplemented with six carbon sugar,- assimilable phosphate yeast extract and alpha pinene at temperature ranging 25-30°C for 6h and recovering the 4,6,6, trimethyl bicyclo (3.1.1) hept 3- en-2-one by known methods such as here in described. 2. An improved process for the production of 4,6,6, trimethyl bicyclo [3.1.1] hept -3-en-2-one using novel penicillium sp. as herein described with reference to the examples.

Full Text

The present invention relates to an improved process for the production of the 4,6,6, trimethyl bicyclo (3.1.1) hept-3en-2one using a novel Penicillium sp. More particularly the present invention relates to a process for production of 4,6,6, trimethyl bicyclo (3.1.1) hept - 3en - 2one having formula 1 as shown in accompanying drawing by biotransformation using a novel Penicillium sp.
Production of food flavours employing microbi
al processes is attractive from the view point of
process economy in the utilisation of inexpensive and
readily available precursor compounds. These are valued
as flavour chemicals and are utilised as food additives.
The conversion assumes commercial importance because the
starting materials are low-cost and yield value added
products.
The production of 4,6,6, trimethyl bicyclo [3.1.1]hept 3-en-2one (fig 1), which is the subject of present study comes from the oil of verbena, dill, spearmint and eucalyptus oil (Guenther.E., The Essential Oils, (1952) vol 2.444. Fritzche Brothers, Inc. New York, Publisher: D.Van Nostrand Company Inc. It is commonly known as verbenone with structural formula as shown in the drawing accompanying this specification.
Leaves, the natural source, are mainly sold for
the herb and drug trade, and small quantities are used for distillation of the essential oil. The distillation process gives a very small yield of the oil generally varying from 0.072% to 0.19% oil. By the present method it can be produced at economical price and would be a valuable adjunct to food and can be used to great advantage in certain types of flavours. The current market price of 4,6,6, trimethyl bicyclo [3.1.1] hept-3en-2one is about Rs.80,000 per kg. It can be formed through microbial source, via hydrogenated bicyclic monoterpene hydrocarbon as shown by Ramadevi, J. and Bhattacharyya. P.K. (1978). Molecular rearrangements in the microbiological transformations of terpenes and chemical logic of microbial process. J. Ind. Chem. Soc, Vol.IV, 1131-1137. The monoterpene alpha-pinene is a by product in lime peel oil, is mainly a waste product from food processing industries. It is also available from turpentine oil as a by product. The utilisation of the low cost substrate to 4,6,6, trimethyi bicycio [3.1.1] hept-3en-2one, a nign value and high priced flavour compound is utilised by a bakeries, confectionary, ice cream and soft drink industry. The compound itself does not have any nutritional value, however, it increases the flavour of the food items.
Recently, this subject has received a lot of attention. Although work is being done abroad on similar lines of biotransformation (Nakayama. H, 1978; Cadwallader, 1989; Cheetham, 1995) Nakayama. H, Yamaguchi. Y,
Machida, Iwasaki.S, Komtsu.A and Shinoda.A D-L-Menthol
(1978) Japan. Kokai Source: CA , 8 9 , 7 4217K.
Cadwallader.K.R, Braddock.R.J, Parish.M.E and Higgins.D.P Bioconversion of (+) limonene by Pseudomo-nas gladioli, (1989) Journal of Food Science vol 54 1241-45. Cheetham Peter.S.J. Biotransformations; New Routes to food ingredients. (1995) Chemistry and Industry 3 April. There is no patent or process developed for biotransformation of this particular compound using Penicillium sp.
There is no work of similar kind being carried out. Therefore, there is an urgent need to have this type of a process which will help in the utilisation of monoterpene hydrocarbon alpha pinene, a by propduct from the waste of the food industries.
A few (four) patents exist on the production of
verbenol, 4,6,6-trimethyl bicyclo [3.1.1]hept-2-en-2-ol
utilising yeast and bacterial cultures. The patent
(No. Int. Pat. Class: C07c-035/28;C 12N-001/26:C12P-
007/02.) by Repph et al show the microbial oxidation
from sugar acids to 4, 6, 6 -trimethy1
bicyclo[3.1.1]hept-2-en-2-ol verbenol, but not to
bicyclo- [3.1.1] hept-3en-2-one , verbenone utilising
Acetobacter, Gluconobacter, Pseudomonas, Aspergillus
and Penicillium. In his patent, Repph et al. has
mentioned the formation of 4,6,6, trimethyl bicyclo
[3.1.1] hept-3en-2-one along with major component 4,6,6- trimethyl bicyclo [3.1.1] hept- 2-en-2-ol utilising methylotrophi Hansenula polvmorpha strain.
There is no patent till date for the production of 4,6,6, trimethyl bicyclo- [3.1.1] hept- 3-en-2one from bicyclic monoterpene hydrocarbon utilizing Penicillium Sp.
The main objective of the invention is to produce verbenone a value flavour compound which has wide usage in food industries and used as a food flavour additive starting from alpha pinene a by product from lime peel oil which is a waste from food industries.
The other object of the present invention is to produce verbenone using a novel strain of Penicillium.
Another object is to attain high yeild . Still another object is to supplement the nutrient medium with special components to obtain a high yield.
Accordingly the present invention provides an improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in
described in a potato dextrose broth medium supplemented
with six carbon sugar, assimilable phosphate yeast
extract and alpha pinene at temperature ranging 25-
30°C for 6h and recovering the 4,6,6, trimethyl
bicyclo (3.1.1) hept 3- en-2-one by known methods such as here in described.
In the persent investigation the compound 4,6,6, trimethyl bicyclo (3.1.1) hept -3-en-2-one has been produced starting from bicyclic monoterpene hydrocarbon. The production for the maximum yield has been optimised and media composition has also been varied to obtain the desired results. Concentrations of carbon, yeast extract and phosphorus were shown to affect the yield to a large extent. Microbial transformation specially for the production of food flavours is a recent technique. Earlier some workers have tried biotransformation studies using the substrate alpha pinene as sole source of carbon but in the present study the substrate was varied over a range and it was found that after certain limit the substrate is toxic to the cells and therefore it has been selected by its capacity for the conversion under appropriate growth conditions. The organism was isolated from the soil collected from different areas. The organism isolated from the soil under the lime tree had a higher potency i.e. capacity for bioconversion, therefore, the organism were selected and isolated from the soil under the
lime tree where expectedly the microorganisms had adapted themselves to grow on monoterpene hydrocarbons.
The underlining principle in the present invention is the use of Surface Response Method for obtaining the maximum conversion yield of 15-2 0%. In the present invention it has been possible to reduce the time of conversion (6 h) , where -as earlier studies indicate a much as 40 to 44 hours. Ramadevi.J and Bhattacharyya.P.K Microbiological transformations of terpenes: part XXIII fermentation of geraniol, nerol and limonene by a soil pseudomonad, Pseudomonas incognita (linalool strain) Ind. J. Biochem. and Biophys. 14, 1977, 288-91. Similarly the yield of the flavouring compounds reported in the patents on conversion of bicyclic monoterpene hydrocarbn to bicyclo- [3.1.1] hept-3-en-2-one has been (10%) reported in the patents on the conversion of bicyclo monoteropene hydrocarbon only by an yeast isolate, Hansenula Repph.H.D Stottme-ist.V ,Duresch.R, Richter.H.P, Schmidt.J, Dorr.M, Hanfe.G, Weber.L (1988) Int. patent.
At present there is no process available as mentioned here. It is, therefore, felt that the present data is of great importance or advantage for the food industries.
The novelty of the invention lies in use of Peni-cillium sp. for biotransformation of alpha pinene to 4,6,6, trimethyl bicylo (3.1.1) hept 3en-2one and maxi mising the yield much more than which is produced by the plant sources or by any other natural source being used in hither to known processes.
Today, there is a lot of awareness especially for food additives like the use of natural flavours and colorants as, more and more people are going in for natural flavours. To produce flavours through microbial source is also natural process.
The invention is related to a use of novel strain of organism being used to produce a product,by a bio-technological process. The culture has been deposited in CFTRI culture collection centre, and designated as CFR 201203 dated 10-4-96.
For the bioconversion, the culture was isolated from the soil. This was subsequently serially diluted and plated on potato dextrose agar medium. The plates were incubated at 25° to 35°C for a period of 3-10 days. The spores were taken for the experiment.
The isolated strain of penicillium sp has the following characteristics the spores are greyish in colour, grows well on potato dextrose agar at 3 0°C the
spores have circular margins. The pigment produced slightly diffuses into the agar medium.
Hence the present invention provides a biotechno-logical process mediated by a selected strain of Peni-cillium for the production of 4,6,6, trimethyl bicy-clo- [3.1.1]hept-3-en-2- one, generally described as below:
A process for the production of 4,6,6, trimethyl bicyclo- [3.1.1]hept-3-en-2one was obtained by modifying the medium and supplementing it with additional six carbon sugar, phosphate and yeast extract.
The process comprises of the following steps.
1. The culture is maintained on Potato dextrose agar
at 30°C
2. Greyish spores were taken in potato dextrose broth
at 3 0°C and grown for 16.5 h.
3. The grown cells were incubated with substarate alpha pinene for 6 h at 120 rpm at 30°C in phosphate buffer at pH 7.0.
4 . The product was extracted in dichloromethane and
analyzed on GC.
The invention is further illustrated by the following examples, which should not be construed to limit the scope of the invention.
Example-1
The spore suspension was taken in potato dextrose broth with an initial cell density 3.0 O.D on spectrophotometer. This was incubated at 30°C, 120 rpm and taken in buffer at pH 7.0 With addition of substrate alpha pinene. The percent yield conversion was 3.2% or 640 ug/ 100ml to the desired product. The product was confirmed on gas chromatography along with the standard and also by gas chromatography mass spectrophotometer fragmentation pattern. GC analysis was done on a Shimadzu Gas chromatograph equipped with a stainless steel SE-30 column and FID detector. For GCMS (gas chromatography-mass spectrophotometry) data, gas Chromatograph mass spectrophotometer Hewlett-Packard 5995 was used with an HP-20 M (carbowax 20M) column (25m x0.32mm) temperature programme at 60°C (6)-4°C/min to 250°C, C15°C, the gas flow rate: lml/min). 4,6,6, trimethyl bicyclo [3.1.1]hept 3-en-2one content was calculated on the area obtained on gas chromatograph of the samples against the area of the standard compound and characterised by GCMS fragmentation pattern.
Example-2
The spore suspension with initial cell density of 3.0 O.D was taken in potato dextrose medium supplemented with glucose 7% and after incubation at 30°C was taken up in buffer at pH 7.0 with the substrate. The percent yield conversion increased with such cells to 3.36% or 672 ug/100 ml. The increase in glucose concentration supports the product formation.
Example-3
The above spore suspension with additional glucose
in the broth was also supplemented with additional
inorganic phosphate KH2P04(500mg/l) and yeast extract
250mg/l.The cell pellet was taken up in phosphate
buffer with pH 7.0 incubated with substrate alpha
pinene for 6 h. This resulted in increasing the
yield conversion from 15 to 20% (3000 -4000ug/ 100ml) . Additional glucose, KH2P04 and also yeast extract enhanced the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en- 2-one .
The present invention provides a process for the production of 4,6,6, trimethyl bicyclo - [3.1.1] hept-3-en-2-one with number of advantages.
1. A fungal culture that utilises a monoterpene hydro
carbon isolated from the waste product of essential oil
industry has been isolated.
2. This organism is capable of converting low cost
essential oil constituent of citrons oil in to a high
valued flavouring compound that has demand in the food
processing industry.
3. There is no fungal culture known to our knowledge
till date producing bicyclo - {3.1.1} hept - 3 - en - 2
- one.

We Claim:
1. An improved process for the production of 4,6,6, trimethyl bicyclo (3.1.1) hept 3-en-2-one using novel Penicillium sp. which comprises growing a novel strain of Penicillium sp. having characteristics as here in described in a potato dextrose broth medium supplemented with six carbon sugar,- assimilable phosphate yeast extract and alpha pinene at temperature ranging 25-30°C for 6h and recovering the 4,6,6, trimethyl bicyclo (3.1.1) hept 3- en-2-one by known methods such as here in described.
2. An improved process for the production of 4,6,6, trimethyl bicyclo [3.1.1] hept -3-en-2-one using novel penicillium sp. as herein described with reference to the examples.

Documents:

381-del-1998-abstract.pdf

381-del-1998-claims.pdf

381-del-1998-complete specification [granted].pdf

381-del-1998-correspondence-others.pdf

381-del-1998-correspondence-po.pdf

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

381-del-1998-drawings.pdf

381-del-1998-form-1.pdf

381-del-1998-form-2.pdf

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Patent Number

186980

Indian Patent Application Number

381/DEL/1998

PG Journal Number

51/2003

Publication Date

22-Dec-2001

Grant Date

02-Aug-2002

Date of Filing

13-Feb-1998

Name of Patentee

DEPARTMENT OF BIOTECHNOLOGY, GOVERNMENT OF INDIA.

Applicant Address

BLOCK2, (7TH & 8TH FLOOR),CGO COMPLEX, LODI ROAD, NEW DELHI-110003, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	RENU AGARWAL	DEPARTMENT OF FOOD MICROBIOLOGY, CFTRI, MYSORE, INDIA
2	NAZHAT-UL-AINN	DEPARTMENT OF FOOD MICROBIOLOGY, CFTRI, MYSORE, INDIA
3	RICHARD JOSEPH	DEPARTMENT OF FOOD MICROBIOLOGY, CFTRI, MYSORE, INDIA

PCT International Classification Number

N/A

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA