Title of Invention	THREE STAGE BIOMETHANATION OF PROTEIN RICH FEEDS
Abstract	The invention presented is a three stage biomethanation process for conversion of protein rich feeds into a combustible mixture of methane and carbon dioxide (biogas). The overall retention time for biomethanation is reduced by pre- treatment of protein rich feeds, which could be by physical, chemical and/or enzymatic methods. The process brings about faster conversion of protein rich biomass into biogas in three stages namely hydrolysis, acidogenesis and methanogenesis. The pre- treatment step hastens the hydrolysis process which substantially reduces the hydraulic retention time and increases the cost effectiveness of the plant. The biogas generated has applications as kitchen fuel, electrical power generation or transportation fuel from renewable biomass and in a cost effective manner.

Title of Invention

THREE STAGE BIOMETHANATION OF PROTEIN RICH FEEDS

Abstract

The invention presented is a three stage biomethanation process for conversion of protein rich feeds into a combustible mixture of methane and carbon dioxide (biogas). The overall retention time for biomethanation is reduced by pre- treatment of protein rich feeds, which could be by physical, chemical and/or enzymatic methods. The process brings about faster conversion of protein rich biomass into biogas in three stages namely hydrolysis, acidogenesis and methanogenesis. The pre- treatment step hastens the hydrolysis process which substantially reduces the hydraulic retention time and increases the cost effectiveness of the plant. The biogas generated has applications as kitchen fuel, electrical power generation or transportation fuel from renewable biomass and in a cost effective manner.

Full Text	FORM 2 THE PATENT ACT 1970 & The Patents Rules, 2003 PROVISIONAL/ SPECIFICATION (See section 10 and rule 13) 1. TITLE OF THE INVENTION: "Three Stage Biomethanation of Protein Rich Feeds' 2. APPLICANT (a) NAME: (b) NATIONALITY: (c) ADDRESS: 1. Kirloskar Oil Engines Limited 2. Gangotree Eco Technologies Pvt. Ltd. Indian Companies registered under the provisions of the Companies Act,1956 1. Laxmanrao Kirloskar Path, Khadki, Pune 411003, M. S. India. 2. Flat no.l Susheeldatta Apts., Plot No.4, Padamarekha Society Karvenagar, Pune 411052, M. S., India. 3. PREAMBLE TO THE DESCRIPTION PROVISIONAL The following specification describes the invention. COMPLETE The following specification particularly describes the invention and the manner which it is to be performed. Technical field: The present invention relates to a three stage biomethanation process. More particularly, it relates to a three stage biomethanation process wherein a commercially available biomass that is rich in proteins is converted into a combustible mixture of methane and carbon dioxide using microbial consortia. Background of the invention & the Prior art: With the supply of fossil fuels getting progressively scarce there is an increasing trend to generate energy from renewable sources. One of the most popular routes followed today is conversion of grains into ethanol. This route has two drawbacks. One is that it threatens availability of food for human beings. The other is that the process utilizes only the carbohydrate component of the grains and not the protein component. An alternate route of converting biomass into biogas opens up the possibility of utilizing non-edible biomass for generation of energy. Research work is available in published literature on conversion of a variety of non-edible feed stocks into biogas. This route can make use of protein component of the biomass as well for the generation of biogas. In order to bring about the conversion of a protein into methane, it has to be taken through a number of steps. Proteins may exist in a combined form (such as glycoproteins, lipoproteins and the like) or in a free form. Initially all these have to be converted into a free form. The proteins are not present in a soluble form at their iso-electric pH and their solubilization may need acidity adjustment. These then have to be de-polymerized before the constituent amino acids are able to enter bacterial cells so that the further reactions of acidogenesis and methanogenesis can take place leading to generation of methane. When a microbial cell is starved, it synthesizes a number of agents to bring about sequential conversion of available proteins in the medium into smaller sub units (or even all the way to free amino acids) that can cross the bacterial cell wall. Many times, the synthesis of such agents by the microbial cell ceases when starvation is over as a result of entry of food inside the cell wall. The cycle then keeps repeating. 1) BIAF: Bharatiya Agro Industries Foundation (BIAF) at Pune, India has published a report stating: that they have succeeded in adding progressively increased quantities of castor cake to cattle dung anci observed an increase in the quantity of biogas generated. As the nitrogen of the castor cake protein is recoverable in the biogas slurry, there is realization of energy without sacrificing its fertilizer value and the process becomes attractive. Drawback: The bioconversion process is slow, essentially because of the slow hydrolysis. Because of the limited hydrolyzed protects, the entry of degraded molecules in the microbial cell is limited. 2) WO 2007052306- A THREE-STEP BIOMETHAN TION PROCESS A three step biomethanation process, to convert starch or sugary agricultural feed stock into a methane rich gas mixture for facilitating generation of biogas to be used as kitchen fuels, electrical power or transportation from renewable biomass in a cost-effective manner forms the basis of this patent. The process brings about conversion of starch-rich or sugar-rich biomass into methane through three stages, namely, hydrolysis, acidogenesis and methane formation. The present invention deploys enzymes/ physical/ microbial parameters to hasten the hydrolytic reaction) of Carbohydrates in the first stage. It also deploys microbial consortia that have been enriched for a targeted feed in order to speed up the process of this conversion. The formation of alcohol from starch is prevented, as a result of the consortia deployed, the conversion efficiency is substantially improved and the retention time is also reduced to cut the capital cost of the plant. Drawback: The process described herein is used for converting only starch or sugar rich feeds for the conversion into the biogas. The hydrolysis of the protein component continues to be a bottle neck for the overall reaction. 3) WO 2003066664- COMPLEX TECHNOLOGIES USING ENZYMATIC PROTEIN HYDROLYSIS The present invention comprises various uses of enzymatic protein hydrolysate ('EPH'). The EPH production process involves the profound enzymatic protein hydrolysis of marine animal biomass, whereby cold-water fish wastes are reacted with viscera containing aggressive enzymes that are effective in even mild (slightly alkaline) medium of water. The EPH may be dried to a powder state that is made of approximately 70 % - 90 % free amino acids, 10 % - 20 % highly molecular peptides and 3 % - 5 % vitamins, minerals and oils. The biological value of the EPH stems from the special mix of nutrients and the method by which they are obtained. EPH is particularly. suitable as an enhancer for the acceleration of known biological and industrial processes, and most particularly, processes depending upon bacterial or cellular action. The low cost of this highly nutritious product makes it possible, to utilize mixtures of amino acids and other nutrients in bio-industrial processes to make them cost-competitive with competing processes. Drawback: The patent claims that proteins from fish or other biomass, when hydrolyzed, gives rise to a mixture of amino acids that are nutritionally important in microbial fermentations. No attempt has been made to use protein rich biomass as a feed for biogas production. Other Drawbacks in Prior art: 1. Protein feed may contain some toxins, which are potent. The hazardous effects of such proteins are needed to overcome. Example: Castor cake contains toxin ricin, which is protein in nature. 2. Degradation of proteins to its sub units or amino acids becomes the limiting factor in their conversion where a complex protein containing feeds are converted to biogas in a digester. This results in long retention times for the conversion (above 30 days) and hence the plants need larger digester volumes that increase the capital cost. 3. In case of generation of biogas as a commercial source of energy, the use of mixture of enzymes is not economically feasible. Object of the invention: The main object of the present invention is to provide a method for pretreatment of protein containing feed, which is used for generation of biogas. Another object of the present invention is to accelerate the process of biomethanation so as to reduce the hydraulic retention time of the conventional process of biomethanation of protein rich feeds. Yet another object of the present invention is to provide a fast, economic and efficient process for pretreatment of protein rich biomass. Yet another object of the present invention is to reduce the capital cost of the equipment. Detailed description of present invention: The foregoing objects of the invention are accomplished and the problems and shortcomings associated with prior art techniques and approaches are overcome by the present invention described in the present embodiment. Protein rich feeds, when suitably pre-treated using physical methods such as size reduction, pH adjustment and/or moist heat, result in making them amenable to faster breakdown. Such feeds when subjected to pre-treatment with chemical agents and proteolytic enzymes (or deploying microbial population known to bring about proteolysis by exoenzymes released by them), either effected singly or in conjunction with physical pre-treatment mentioned above, results in making Such feeds more amenable to faster conversion into biogas. Such pre-treattrient results in substantially reduced residence time for the feed in the reactor and a much faster conversion to biogas resulting in a substantial savings in the capital cost of the plant. Advantages of the present invention: • The rate limiting step in the conversion of protein rich feeds during biomethanation is the degradation of proteins into small molecules enough to enter the microbial cell. This is normally brought about by enzymes specifically produced by the microbial cells and exported into the medium. • The process described in the invention aims at speeding up the breakdown of proteins by using externally added enzymes. This accelerates the rate limiting step. As the hydraulic retention time is reduced, the biogas plant size also gets reduced. The pre-treatment using physical method and proteolysis as proposed is well-focused/ fast and economically feasible. It results in major savings in the capital cost of the plant. Dated this 30th September 2008, [M. D. BHATE (Agent for the Applicant)

Full Text

FORM 2
THE PATENT ACT 1970
&
The Patents Rules, 2003
PROVISIONAL/ SPECIFICATION
(See section 10 and rule 13)
1. TITLE OF THE INVENTION:
"Three Stage Biomethanation of Protein Rich Feeds'

2. APPLICANT
(a) NAME:
(b) NATIONALITY:
(c) ADDRESS:

1. Kirloskar Oil Engines Limited
2. Gangotree Eco Technologies Pvt. Ltd.
Indian Companies registered under the provisions of the Companies Act,1956
1. Laxmanrao Kirloskar Path, Khadki, Pune 411003,
M. S. India.
2. Flat no.l Susheeldatta Apts., Plot No.4,
Padamarekha Society Karvenagar, Pune 411052, M. S., India.

3. PREAMBLE TO THE DESCRIPTION

PROVISIONAL
The following specification describes the invention.

COMPLETE
The following specification particularly describes the invention and the manner which it is to be performed.

Technical field:
The present invention relates to a three stage biomethanation process. More particularly, it relates to a three stage biomethanation process wherein a commercially available biomass that is rich in proteins is converted into a combustible mixture of methane and carbon dioxide using microbial consortia.
Background of the invention & the Prior art:
With the supply of fossil fuels getting progressively scarce there is an increasing trend to generate energy from renewable sources. One of the most popular routes followed today is conversion of grains into ethanol.
This route has two drawbacks. One is that it threatens availability of food for human beings. The other is that the process utilizes only the carbohydrate component of the grains and not the protein component.
An alternate route of converting biomass into biogas opens up the possibility of utilizing non-edible biomass for generation of energy. Research work is available in published literature on conversion of a variety of non-edible feed stocks into biogas. This route can make use of protein component of the biomass as well for the generation of biogas.
In order to bring about the conversion of a protein into methane, it has to be taken through a number of steps. Proteins may exist in a combined form (such as glycoproteins, lipoproteins and the like) or in a free form. Initially all these have to be converted into a free form. The proteins are not present in a soluble form at their iso-electric pH and their solubilization may need acidity adjustment. These then have to be de-polymerized before the constituent amino acids are able to

enter bacterial cells so that the further reactions of acidogenesis and methanogenesis can take place leading to generation of methane.
When a microbial cell is starved, it synthesizes a number of agents to bring about sequential conversion of available proteins in the medium into smaller sub units (or even all the way to free amino acids) that can cross the bacterial cell wall. Many times, the synthesis of such agents by the microbial cell ceases when starvation is over as a result of entry of food inside the cell wall. The cycle then keeps repeating.
1) BIAF: Bharatiya Agro Industries Foundation (BIAF) at Pune, India has
published a report stating: that they have succeeded in adding progressively
increased quantities of castor cake to cattle dung anci observed an increase in the
quantity of biogas generated. As the nitrogen of the castor cake protein is
recoverable in the biogas slurry, there is realization of energy without sacrificing
its fertilizer value and the process becomes attractive.
Drawback: The bioconversion process is slow, essentially because of the slow hydrolysis. Because of the limited hydrolyzed protects, the entry of degraded molecules in the microbial cell is limited.
2) WO 2007052306- A THREE-STEP BIOMETHAN TION PROCESS
A three step biomethanation process, to convert starch or sugary agricultural feed stock into a methane rich gas mixture for facilitating generation of biogas to be used as kitchen fuels, electrical power or transportation from renewable biomass in a cost-effective manner forms the basis of this patent. The process brings about conversion of starch-rich or sugar-rich biomass into methane through three stages, namely, hydrolysis, acidogenesis and methane formation. The present invention deploys enzymes/ physical/ microbial parameters to hasten the hydrolytic reaction) of Carbohydrates in the first stage. It also deploys

microbial consortia that have been enriched for a targeted feed in order to speed up the process of this conversion. The formation of alcohol from starch is prevented, as a result of the consortia deployed, the conversion efficiency is substantially improved and the retention time is also reduced to cut the capital cost of the plant.
Drawback: The process described herein is used for converting only starch or sugar rich feeds for the conversion into the biogas. The hydrolysis of the protein component continues to be a bottle neck for the overall reaction.
3) WO 2003066664- COMPLEX TECHNOLOGIES USING ENZYMATIC PROTEIN HYDROLYSIS
The present invention comprises various uses of enzymatic protein hydrolysate ('EPH'). The EPH production process involves the profound enzymatic protein hydrolysis of marine animal biomass, whereby cold-water fish wastes are reacted with viscera containing aggressive enzymes that are effective in even mild (slightly alkaline) medium of water. The EPH may be dried to a powder state that is made of approximately 70 % - 90 % free amino acids, 10 % - 20 % highly molecular peptides and 3 % - 5 % vitamins, minerals and oils. The biological value of the EPH stems from the special mix of nutrients and the method by which they are obtained. EPH is particularly. suitable as an enhancer for the acceleration of known biological and industrial processes, and most particularly, processes depending upon bacterial or cellular action. The low cost of this highly nutritious product makes it possible, to utilize mixtures of amino acids and other nutrients in bio-industrial processes to make them cost-competitive with competing processes.
Drawback: The patent claims that proteins from fish or other biomass, when hydrolyzed, gives rise to a mixture of amino acids that are nutritionally important in microbial fermentations. No attempt has been made to use protein rich biomass as a feed for biogas production.

Other Drawbacks in Prior art:
1. Protein feed may contain some toxins, which are potent. The hazardous effects of such proteins are needed to overcome. Example: Castor cake contains toxin ricin, which is protein in nature.
2. Degradation of proteins to its sub units or amino acids becomes the limiting factor in their conversion where a complex protein containing feeds are converted to biogas in a digester. This results in long retention times for the conversion (above 30 days) and hence the plants need larger digester volumes that increase the capital cost.
3. In case of generation of biogas as a commercial source of energy, the use of mixture of enzymes is not economically feasible.
Object of the invention:
The main object of the present invention is to provide a method for pretreatment of protein containing feed, which is used for generation of biogas.
Another object of the present invention is to accelerate the process of biomethanation so as to reduce the hydraulic retention time of the conventional process of biomethanation of protein rich feeds.
Yet another object of the present invention is to provide a fast, economic and efficient process for pretreatment of protein rich biomass.
Yet another object of the present invention is to reduce the capital cost of the equipment.

Detailed description of present invention:
The foregoing objects of the invention are accomplished and the problems and shortcomings associated with prior art techniques and approaches are overcome by the present invention described in the present embodiment.
Protein rich feeds, when suitably pre-treated using physical methods such as size reduction, pH adjustment and/or moist heat, result in making them amenable to faster breakdown. Such feeds when subjected to pre-treatment with chemical agents and proteolytic enzymes (or deploying microbial population known to bring about proteolysis by exoenzymes released by them), either effected singly or in conjunction with physical pre-treatment mentioned above, results in making Such feeds more amenable to faster conversion into biogas.
Such pre-treattrient results in substantially reduced residence time for the feed in the reactor and a much faster conversion to biogas resulting in a substantial savings in the capital cost of the plant.
Advantages of the present invention:
• The rate limiting step in the conversion of protein rich feeds during biomethanation is the degradation of proteins into small molecules enough to enter the microbial cell. This is normally brought about by enzymes specifically produced by the microbial cells and exported into the medium.
• The process described in the invention aims at speeding up the breakdown of proteins by using externally added enzymes. This accelerates the rate limiting step. As the hydraulic retention time is reduced, the biogas plant size also gets reduced.

The pre-treatment using physical method and proteolysis as proposed is well-focused/ fast and economically feasible. It results in major savings in the capital cost of the plant.
Dated this 30th September 2008,
[M. D. BHATE (Agent for the Applicant)

Documents:

2134-MUM-2008-ABSTRACT(6-10-2009).pdf

2134-MUM-2008-ASSIGNMENT(29-9-2009).pdf

2134-MUM-2008-CLAIMS(6-10-2009).pdf

2134-MUM-2008-CLAIMS(AMENDED)-(17-9-2013).pdf

2134-MUM-2008-CLAIMS(MARKED COPY)-(17-9-2013).pdf

2134-MUM-2008-Claims-110215.pdf

2134-MUM-2008-CORRESPONDENCE(29-9-2009).pdf

2134-MUM-2008-CORRESPONDENCE(6-8-2010).pdf

2134-mum-2008-correspondence.pdf

2134-MUM-2008-DESCRIPTION(COMPLETE)-(6-10-2009).pdf

2134-mum-2008-discription(provisional).doc

2134-mum-2008-discription(provisional).pdf

2134-mum-2008-form 1.pdf

2134-MUM-2008-FORM 13(17-9-2013).pdf

2134-MUM-2008-FORM 18(16-9-2010).pdf

2134-mum-2008-form 2(6-10-2009).pdf

2134-MUM-2008-FORM 2(TITLE PAGE)-(6-10-2009).pdf

2134-MUM-2008-FORM 2(TITLE PAGE)-(PROVISIONAL)-(6-10-2008).pdf

2134-mum-2008-form 2(title page).pdf

2134-mum-2008-form 2.doc

2134-mum-2008-form 2.pdf

2134-MUM-2008-FORM 3(6-10-2009).pdf

2134-mum-2008-form 3.pdf

2134-MUM-2008-FORM 5(6-10-2009).pdf

2134-mum-2008-form 6(29-9-2009).pdf

2134-MUM-2008-GENERAL POWER OF AUTHORITY-110215.pdf

2134-MUM-2008-MARKED COPY-110215.pdf

2134-MUM-2008-OTHERS-110215.pdf

2134-mum-2008-power of attorney.pdf

2134-MUM-2008-POWER OF AUTHORITY(6-10-2008).pdf

2134-MUM-2008-REPLY TO EXAMINATION REPORT(17-9-2013).pdf

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Patent Number

265757

Indian Patent Application Number

2134/MUM/2008

PG Journal Number

12/2015

Publication Date

20-Mar-2015

Grant Date

12-Mar-2015

Date of Filing

06-Oct-2008

Name of Patentee

KIRLOSKAR OIL ENGINES LIMITED

Applicant Address

LAXMANRAO KIRLOSKAR PATH KHADKI, PUNE

Inventors:

#	Inventor's Name	Inventor's Address
1	GONDHALEKAR, SANTOSH RAGHUNATH	SHREE BUNGALOW, PLOT NO. 22, TUKARAM SOCIETY, SANT NAGAR, PUNE 411009,
2	JOSHI, ARVIND PURUSHOTTAM	FLAT NO. 3, RACHANA CLASSIC, GULMOHOR PARK, ITI ROAD, AUNDH, PUNE 411007,
3	MATE, NITANT VISHNU	9, 'SHRIMAN' 793, BHANDARKAR ROAD, PUNE 411004,

PCT International Classification Number

B01J41/04; C02F3/28; B01J41/00; C12P5/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA