WO2018136017A1 - Method for increasing methane yield in anaerobic digesters with a cellulolytic bacteria cocktail (cebac-s) prepared with rumen fluid obtained from sheep - Google Patents
Method for increasing methane yield in anaerobic digesters with a cellulolytic bacteria cocktail (cebac-s) prepared with rumen fluid obtained from sheep Download PDFInfo
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- WO2018136017A1 WO2018136017A1 PCT/TR2017/050034 TR2017050034W WO2018136017A1 WO 2018136017 A1 WO2018136017 A1 WO 2018136017A1 TR 2017050034 W TR2017050034 W TR 2017050034W WO 2018136017 A1 WO2018136017 A1 WO 2018136017A1
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- cebac
- digesters
- cocktail
- cellulolytic bacteria
- bacteria cocktail
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Classifications
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
- C02F11/04—Anaerobic treatment; Production of methane by such processes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
- C12P5/023—Methane
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Definitions
- This invention is related to a method of increasing methane yield in anaerobic digesters treating lignocellulosic biomass with a cellulolytic bacteria cocktail (cebac-s).
- this invention is related to a method for increasing methane yield in anaerobic digesters treating lignocellulosic biomass such as animal manure, agricultural waste, energy plants and food residue, sewage sludge, household wastewater and industrial wastewater, by adding a cellulolytic bacteria cocktail (cebac-s) by increasing the hydrolysis efficiency at a maximum level which also stimulates methane production.
- lignocellulosic biomass such as animal manure, agricultural waste, energy plants and food residue, sewage sludge, household wastewater and industrial wastewater
- a cellulolytic bacteria cocktail cebac-s
- Anaerobic digesters enable to obtain energy from many biodegradable waste such as animal manure, agricultural waste, and sewage sludge, industrial and household waste.
- hydrolysis phase limits speed during the anaerobic biotransformation of lignocellulose-rich wastes. The recalcitrant characteristic of the biomass makes the hydrolysis of these biomasses more difficult.
- the aim of the invention is to carry out the method for increasing methane yield in anaerobic digesters by a cellulolytic bacteria cocktail (cebac-s) in order to increase the hydrolysis rate and efficiency in anaerobic digesters during the treatment of lignocellulose-rich wastes.
- a cellulolytic bacteria cocktail cebac-s
- Another aim of the invention is to carry out a method to increase methane yield in anaerobic digesters, comprising Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes and Synergistales.
- anaerobic digesters comprising Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes and Synergistales.
- methane production can be increased significantly, by the addition of a small amount such as 4% of the specific cellulolytic bacteria cocktail (Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, Synergistales) that has been developed.
- the bacteria cocktail that has been added has improved the hydrolysis phase of wheat straw which is a lignocellulosic waste, and has increased volatile acid production, and also increased methane production.
- specific cellulolytic bacteria cocktail accelerates the hydrolysis phase.
- the invention is a method to increase methane yield in anaerobic digesters comprising cellulolytic bacteria cocktail (cebac-s) which comprises the steps of: obtaining a rumen fluid from a healthy sheep which has not taken any antibiotics, - obtaining a cellulolytic bacteria cocktail by inoculation of the obtained
- Rumen fluid consisting of Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, and Synergistalesbacteriaspecies in a medium adding the obtained specific cellulolytic bacteria cocktail into digesters.
- the cellulolytic activities of rumen bacteria provides an advantage during the treatment of lignocellulolytic waste in anaerobic digesters such as animal manure, agricultural waste, energy plants, food residue, sewage sludge, household wastewater and industrial wastewater.
- Environmental parameters such as pH, temperature, redox potential, show similarities between anaerobic digesters and rumen environment. For this reason, the microorganisms found within the rumen eco-system can stay alive in anaerobic digesters which is an engineering system.
- Cellulolytic bacteria have been enriched and a specific cellulolytic bacteria cocktail (cebac-s) has been prepared, according to the method subject of the invention, with serial culturing methods using a special medium inoculated with the rumen fluid obtained from a 10 year old healthy sheep, weighing 85kg' s which has not taken any antibiotics.
- cebac-s specific cellulolytic bacteria cocktail
- the rumen fluid bacteria composition obtained from the sheep comprises bacterial species of 74% Bacteroidetes, 15%Firmicutes, 3% Anaeroplasma, 3% Fibrobacter, l%Cyanobacteria, and 4% other bacteria.
- the rumen fluid has been inoculated into the DSMZ 1036 medium that has been modified for this method and serial enriching studies have been carried out.
- the content of the specific cellulolytic bacteria cocktail (cebac-s) that has been obtained is as follows: at least 24% Bacteroidales, at least 17% Lachnospiraceae, at least 15% Lentisphaeria, at least 14%) Ruminococcaceae, and at least 14% Firmicutes and at least 9% Synergistales.
- the ratios that have been provided are the abundance of the bacteria species inside biomass. According to an application of the invention, in the case that the bacteria species used have the lowest possible percentage ratio, the mixture may comprise different types of bacteria besides said bacteria.
- methane production can be increased significantly, by the addition of a small amount such as 4% of the total microbial community of the digesters, of the specific cellulolytic bacteria cocktail into anaerobic digesters.
- the bacteria cocktail that has been added has improved the hydrolysis phase of wheat waste which are lignocellulosic biomass, and has increased volatile acid production, and therefore improvement in methane production has also been obtained.
- the findings that have been obtained have shown that specific cellulolytic bacteria cocktail accelerates the hydrolysis phase.
Abstract
This invention is related to a method for increasing methane yield in anaerobic digesters by adding a cellulolytic bacteria cocktail (cebac-s) in order to increase methane yield in a system by increasing the hydrolysis efficiency at a maximum level, in anaerobic digesters that are processed with lignocellulosic waste, such as animal manure, agricultural waste, energy plants, food residue, sewage sludge, household wastewater and industrial wastewater. The invention is related to a method for increasing methane yield in anaerobic digesters containing a cellulolytic bacteria cocktail (cebac-s), characterized in that said method comprises the steps of obtaining rumen fluid from a healthy sheep that have not been given antibiotics, said fluid comprising Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, Synergistales and other bacteria in order to obtain a cellulolytic bacteria cocktail, and adding said cellulolytic bacteria cocktail to digesters.
Description
DESCRIPTION
METHOD FOR INCREASING METHANE YIELD IN ANAEROBIC DIGESTERS WITH A CELLULOLYTIC BACTERIA COCKTAIL
(CEBAC-S). Technical Field
This invention is related to a method of increasing methane yield in anaerobic digesters treating lignocellulosic biomass with a cellulolytic bacteria cocktail (cebac-s).
More particularly, this invention is related to a method for increasing methane yield in anaerobic digesters treating lignocellulosic biomass such as animal manure, agricultural waste, energy plants and food residue, sewage sludge, household wastewater and industrial wastewater, by adding a cellulolytic bacteria cocktail (cebac-s) by increasing the hydrolysis efficiency at a maximum level which also stimulates methane production. Prior Art
Nowadays, the tendency for renewable energy sources has started rapidly in worldwide. Due to the fuel sources are not environmentally friendly, have become scare and more expensive, this progress has accelerated. One of these renewable energy sources is a-biomass energy. The scientific studies that have been carried out on anaerobic digesters treating lignocellulosic waste (such as animal manure, agricultural waste, energy plants, food residue, and sewage sludge, industrial and household waste) are generally focused on the optimization of operation conditions and increasing of methane yield. Nearly all of the limited number of facilities in our country cannot be operated with a hundred percent efficiency. In the studies that have been conducted, anaerobic digesters have generally been inoculated with animal
manure. Besides this, in laboratory scale studies where the rumen fluid is used as an inoculum, it has been determined that the rumen fluid had a positive effect on hydrolysis yield and therefore it increased methane yield. In these studies a distinctive lignocellulosic microbial community mixture was not obtained. Anaerobic digesters enable to obtain energy from many biodegradable waste such as animal manure, agricultural waste, and sewage sludge, industrial and household waste. However the most significant obstacle in energy production from anaerobic digesters is the hydrolysis phase which limits speed during the anaerobic biotransformation of lignocellulose-rich wastes. The recalcitrant characteristic of the biomass makes the hydrolysis of these biomasses more difficult.
The International patent document that has been filed on 02.04.2009 numbered WO2010114481 Al of the known state of the art, describes a method which accelerates methane production during an anaerobic digestion process. According to this method, glycerol-digestive microbes or pentose is added to anaerobic digesters and they substantially convert biogas into methane.
The Chinese patent document that has been filed on 18.09.2012 numbered CN102876725 A of the known state of the art, describes a method which comprises an active agent that is added to anaerobic processes in order to develop methane fermentation. This active agent which comprises magnesium, copper, nickel and iron, accelerates the anaerobic fermentation process of methane even more.
The German patent document that has been filed on 12.02.2002 numbered DEI 0300082 Al of the known state of the art, describes a simpler construction of recycling from biological wastewater by the addition of a mixture into an anaerobic digestion procedure, wherein said mixture is formed of sulphate, manganese, chloride, and chromium.
However the applications mentioned in these documents are not sufficient enough to increase methane production and hydrolysis yield at the desired amounts inside anaerobic digesters that are being processed with lignocellulolytic waste. For this reason the need to develop the method of increasing methane yield in anaerobic digesters witha cellulolytic bacteria cocktail (cebac-s) subject to the invention has risen.
Objects and Brief Description of the Invention
The aim of the invention is to carry out the method for increasing methane yield in anaerobic digesters by a cellulolytic bacteria cocktail (cebac-s) in order to increase the hydrolysis rate and efficiency in anaerobic digesters during the treatment of lignocellulose-rich wastes.
Another aim of the invention is to carry out a method to increase methane yield in anaerobic digesters, comprising Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes and Synergistales. By means of the method subject to the invention, the hydrolysis rate of lignocellulose-rich wastes has been increased in anaerobic digesters.
It has been proved that methane production can be increased significantly, by the addition of a small amount such as 4% of the specific cellulolytic bacteria cocktail (Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, Synergistales) that has been developed. The bacteria cocktail that has been added, has improved the hydrolysis phase of wheat straw which is a lignocellulosic waste, and has increased volatile acid production, and also increased methane production. Moreover, the findings have shown that specific cellulolytic bacteria cocktail accelerates the hydrolysis phase.
Detailed Description of the Invention
The invention is a method to increase methane yield in anaerobic digesters comprising cellulolytic bacteria cocktail (cebac-s) which comprises the steps of: obtaining a rumen fluid from a healthy sheep which has not taken any antibiotics, - obtaining a cellulolytic bacteria cocktail by inoculation of the obtained
Rumen fluid consisting of Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, and Synergistalesbacteriaspecies in a medium adding the obtained specific cellulolytic bacteria cocktail into digesters. The cellulolytic activities of rumen bacteria provides an advantage during the treatment of lignocellulolytic waste in anaerobic digesters such as animal manure, agricultural waste, energy plants, food residue, sewage sludge, household wastewater and industrial wastewater. Environmental parameters such as pH, temperature, redox potential, show similarities between anaerobic digesters and rumen environment. For this reason, the microorganisms found within the rumen eco-system can stay alive in anaerobic digesters which is an engineering system.
Cellulolytic bacteria have been enriched and a specific cellulolytic bacteria cocktail (cebac-s) has been prepared, according to the method subject of the invention, with serial culturing methods using a special medium inoculated with the rumen fluid obtained from a 10 year old healthy sheep, weighing 85kg' s which has not taken any antibiotics.
It is possible to obtain rumen fluid from animals such as cows, goats, llamas etc. which are referred to as ruminant animals, during alternative applications of the invention. The rumen fluid bacteria composition obtained from the sheep comprises bacterial species of 74% Bacteroidetes, 15%Firmicutes, 3% Anaeroplasma, 3% Fibrobacter, l%Cyanobacteria, and 4% other bacteria. The rumen fluid has been inoculated into the DSMZ 1036 medium that has been modified for this method
and serial enriching studies have been carried out. The content of the specific cellulolytic bacteria cocktail (cebac-s) that has been obtained is as follows: at least 24% Bacteroidales, at least 17% Lachnospiraceae, at least 15% Lentisphaeria, at least 14%) Ruminococcaceae, and at least 14% Firmicutes and at least 9% Synergistales. The ratios that have been provided are the abundance of the bacteria species inside biomass. According to an application of the invention, in the case that the bacteria species used have the lowest possible percentage ratio, the mixture may comprise different types of bacteria besides said bacteria.
In order to obtain maximum performance from anaerobic digesters microorganisms that play a role during the hydrolysis acidogenesis, acetogenesis and methanogenesis steps need to present in the system in certain amounts and they need to function in a symbiotic relationship. The method has been planned within this scope. Besides this, when the ratios to be used were being designed, the applicability thereof in large scale systems, and the competitive relationship thereof with the present microbial community has been taken into consideration. For this reason, the specific cellulolytic bacteria cocktail has been added to digesters at a rate of 2% or 4% of the total microbial community of the digesters. It has been determined that both the rates have a positive effect on methane yield and methane production has increased by %28±5 within the system where 4% of specific cellulolytic bacteria cocktail.
It has been proved that methane production can be increased significantly, by the addition of a small amount such as 4% of the total microbial community of the digesters, of the specific cellulolytic bacteria cocktail into anaerobic digesters. The bacteria cocktail that has been added, has improved the hydrolysis phase of wheat waste which are lignocellulosic biomass, and has increased volatile acid production, and therefore improvement in methane production has also been obtained. Moreover, the findings that have been obtained have shown that specific cellulolytic bacteria cocktail accelerates the hydrolysis phase.
Claims
The invention is a method to increase methane yield in anaerobic digesters withcellulolytic bacteria cocktail (cebac-s) which comprises the steps of: obtaining a rumen fluid from sheep, obtaining cellulolytic bacteria cocktail by inoculating the obtained Rumen fluid comprisingnBacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes, and Synergistalesbacteriaspecies in a medium, adding the obtained cellulolytic bacteria cocktail into digesters.
A method for increasing methane yield in anaerobic digesters with a celluloytic bacteria cocktail (cebac-s) according to claim 1, characterized in that the bacterial composition of the rumen fluid obtained from the sheep comprises 74% Bacteroidetes, 15% Firmicutes, 3% Anaeroplasma, 3%) Fibrobacter, 1% Cyanobacteria and 4% other bacterial types by volume and that the cellulolytic bacteria cocktail comprises at least 24% Bacteroidales, at least 17% Lachnospiraceae, at least 15% Lentisphaeria, at least 14% Ruminococcaceae, at least 14% Firmicutes and at least 9% Synergistalesbacteriatypes by abundance.
A method for increasing methane yield in anaerobic digesters with cellulolytic bacteria cocktail (cebac-s), according to claim 2, characterized in that; the cellulolytic bacteria cocktail comprises 8% different bacteria species by abundance, other than Bacteroidales, Lachnospiraceae, Lentisphaeria, Ruminococcaceae, Firmicutes and Synergistales.
A method for increasing methane yield in anaerobic digesters with a cellulolytic bacteria cocktail (cebac-s), according to claim 1,2 or 3, characterized in that; the cellulolytic bacteria cocktail is added to the digesters at a ratio of 2% or 4% of the total microbial community of the digesters.
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| PCT/TR2017/050034 WO2018136017A1 (en) | 2017-01-20 | 2017-01-20 | Method for increasing methane yield in anaerobic digesters with a cellulolytic bacteria cocktail (cebac-s) prepared with rumen fluid obtained from sheep |
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| PCT/TR2017/050034 WO2018136017A1 (en) | 2017-01-20 | 2017-01-20 | Method for increasing methane yield in anaerobic digesters with a cellulolytic bacteria cocktail (cebac-s) prepared with rumen fluid obtained from sheep |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110699389A (en) * | 2019-11-22 | 2020-01-17 | 大连大学 | Method for producing volatile fatty acid by using rumen microorganisms |
| EP3808850A1 (en) | 2019-07-23 | 2021-04-21 | Recolo, SIA | A method of anaerobic digestion of food waste by novel composition of catalyst |
Citations (6)
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|---|---|---|---|---|
| EP0159054A1 (en) * | 1984-03-09 | 1985-10-23 | Stichting Katholieke Universiteit | A process for producing methane from solid vegetable material |
| DE10300082A1 (en) | 2002-02-12 | 2003-08-21 | Ufl Umweltanalytik Und Forschu | Trace element mixture for increasing the digester gas yield while reducing the amount of organic dry sludge in anaerobic degradation processes involving methane production comprises predetermined number of elements |
| WO2010114481A1 (en) | 2009-04-02 | 2010-10-07 | Agency For Science, Technology And Research | Methods for improving biogas production in the presence of hard substrates |
| US20110111475A1 (en) * | 2009-04-17 | 2011-05-12 | Kuhry Anthony B | Biological/Electrolytic Conversion of Biomass to Hydrocarbons |
| WO2012170989A2 (en) * | 2011-06-09 | 2012-12-13 | Integrated Biochem, Llc | Process of managed ecosystem fermentation |
| CN102876725A (en) | 2012-09-18 | 2013-01-16 | 安徽农业大学 | Active compound additive for promoting fermentation of methane and use method thereof |
-
2017
- 2017-01-20 WO PCT/TR2017/050034 patent/WO2018136017A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0159054A1 (en) * | 1984-03-09 | 1985-10-23 | Stichting Katholieke Universiteit | A process for producing methane from solid vegetable material |
| DE10300082A1 (en) | 2002-02-12 | 2003-08-21 | Ufl Umweltanalytik Und Forschu | Trace element mixture for increasing the digester gas yield while reducing the amount of organic dry sludge in anaerobic degradation processes involving methane production comprises predetermined number of elements |
| WO2010114481A1 (en) | 2009-04-02 | 2010-10-07 | Agency For Science, Technology And Research | Methods for improving biogas production in the presence of hard substrates |
| US20110111475A1 (en) * | 2009-04-17 | 2011-05-12 | Kuhry Anthony B | Biological/Electrolytic Conversion of Biomass to Hydrocarbons |
| WO2012170989A2 (en) * | 2011-06-09 | 2012-12-13 | Integrated Biochem, Llc | Process of managed ecosystem fermentation |
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| Title |
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| LUCAS DANTAS LOPES ET AL: "Exploring the sheep rumen microbiome for carbohydrate-active enzymes", ANTONIE VAN LEEUWENHOEK, vol. 108, no. 1, 1 July 2015 (2015-07-01), DORDRECHT; NL, pages 15 - 30, XP055361678, ISSN: 0003-6072, DOI: 10.1007/s10482-015-0459-6 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3808850A1 (en) | 2019-07-23 | 2021-04-21 | Recolo, SIA | A method of anaerobic digestion of food waste by novel composition of catalyst |
| CN110699389A (en) * | 2019-11-22 | 2020-01-17 | 大连大学 | Method for producing volatile fatty acid by using rumen microorganisms |
| CN110699389B (en) * | 2019-11-22 | 2023-06-27 | 大连大学 | Method for producing volatile fatty acid by utilizing rumen microorganism |
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