# Patent

## Title
Method of stimulating ethanol production and growth of aquatic plants

## Abstract

A method of stimulating ethanol production and growth of aquatic plants includes the steps of placing aquatic plants in a cell containing water and creating an anoxic condition within the pool to initiate an anaerobic process by the aquatic plants. The aquatic plants increase in size and release ethanol by metabolism of stored carbohydrates during the anaerobic process. An oxygenated condition is then created within the cell to initiate an aerobic process. The aquatic plants create and store carbohydrates during the aerobic process. The steps of creating anoxic and oxygenated conditions are repeated to stimulate increased aquatic plant size and to increase the release of ethanol.

## Background
BACKGROUND OF THE DISCLOSURE
Field of the Disclosure
The disclosure relates to ethanol production methods and more particularly pertains to a new ethanol production method for promoting plant growth by plants which produce free ethanol during anaerobic metabolism to form a self-sustaining cycle of plant growth and ethanol production.
SUMMARY OF THE DISCLOSURE
An embodiment of the disclosure meets the needs presented above by generally comprising the steps of placing aquatic plants in a cell containing water and creating an anoxic condition within the pool to initiate an anaerobic process by the aquatic plants. The aquatic plants increase in size and release ethanol by metabolism of stored carbohydrates during the anaerobic process. An oxygenated condition is then created within the cell to initiate an aerobic process. The aquatic plants create and store carbohydrates during the aerobic process. The steps of creating anoxic and oxygenated conditions are repeated to stimulate increased aquatic plant size and to increase the release of ethanol.
There has thus been outlined, rather broadly, the more important features of the disclosure in order that the detailed description thereof that follows may be better understood, and in order that the present contribution to the art may be better appreciated. There are additional features of the disclosure that will be described hereinafter and which will form the subject matter of the claims appended hereto.
The objects of the disclosure, along with the various features of novelty which characterize the disclosure, are pointed out with particularity in the claims annexed to and forming a part of this disclosure.



BRIEF DESCRIPTION OF THE DRAWINGS
The disclosure will be better understood and objects other than those set forth above will become apparent when consideration is given to the following detailed description thereof. Such description makes reference to the annexed drawing wherein:
FIG. 1 is a schematic view of a method of stimulating ethanol production and growth of aquatic plants according to an embodiment of the disclosure.



DESCRIPTION OF THE PREFERRED EMBODIMENT
With reference now to the drawing, and in particular to FIG. 1, a new ethanol production method embodying the principles and concepts of an embodiment of the disclosure and generally designated by the reference numeral 10 will be described.
As illustrated in FIG. 1, the method 10 of stimulating ethanol production and growth of aquatic plants generally comprises harvesting aquatic plants from lakes or ponds which are then introduced into cells. As the method 10 is performed, it may be used to grow and provide aquatic plants as they are needed for future cells or for replacement purposes. The cells are constructed to hold water and may or may not be lined to prevent transfer of fluids and gases into a ground surface supporting the cell. A fine particulate is placed in the cells and the aquatic plants introduced into the cells where they can anchor themselves in the particulate. A fine particulate is used as it may promote less energy expenditure on the part of the aquatic plants to root growth into the particulate and retains a higher percentage of the plant matter above the surface of the particulate.
The number of cells and their size is not crucial to the method and each number and size may be dictated by available land area, access to raw materials and cost controls, though it should be understood that the method may be practiced with a single cell. The cells may have any depth required for the chosen aquatic plant to properly grow. However, it has been found that cell may have a depth of between 10 cm and 7 m to prevent restricted plant growth. The cells may also be temperature controlled and in particular the cell should be prevented from freezing which may kill the aquatic plants. Heat for the cells may be sequestered from waste heat emitted by adjacent ethanol processing plants or any other convenient source of waste heat.
The aquatic plants may be selected from any number of aquatic plants which readily live in or on an aquatic environment such as directly in water or in permanently saturated soil. Further, more than one type of aquatic plant may be used within a single cell. The aquatic plants may include, for example, algae, submersed aquatic herbs such as, but not limited to, Potamogeton pectinatus, Potamogeton crispus, Ruppia maitima, Myriophyllum spicatum, Hydrilla verticillata, Elodea densa, Hippuris vulgaris, Aponogeton boivinianus, Aponogeton rigidifolius, Aponogeton longiplumulosus, Didiplis diandra, Vesicularia dubyana, Hygrophilia augustifolia, Micranthemum umbrosum, Eichhornia azurea, Saururus cernuus, Cryptocoryne lingua, Hydrotriche hottoniiflora, Eustralis stellata, Vallisneria rubra, Hygrophila salicifolia, Cyperus helferi, Cryptocoryne petchii, Vallisneria americana, Vallisneria torta, Hydrotriche hottoniiflora, Crassula helmsii, Limnophila sessiliflora, Potamogeton perfoliatus, Rotala wallichii, Cryptocoryne becketii, Blyxa aubertii and Hygrophila difformmis, duckweeds such as, but not limited to, Spirodela polyrrhiza, Wolffia globosa, Lemna trisulca, Lemna gibba, Lemna minor, and Landoltia punctata, water cabbage, such as but not limited to Pistia stratiotes, buttercups such as but not limited to Ranunculus, water caltrop such as but not limited to Trapa natans and Trapa bicornis, water lily such as Nymphaea lotus, Nymphaeaceae and Nelumbonaceae, water hyacinth such as but not limited to Eichhornia crassipes, Bolbitis heudelotii, and Cabomba, and seagrasses such as but not limited to Heteranthera zosterifolia, Posidoniaceae, Zosteraceae, Hydrocharitaceae, and Cymodoceaceae. Moreover, in one of the various embodiments, a host alga is selected from the group consisting of green algae, red algae, brown algae, diatoms, marine algae, freshwater algae, unicellular algae, multicellular algae, seaweeds, cold-tolerant algal strains, heat-tolerant algal strains, ethanol-tolerant algal strains, and combinations thereof.
The aquatic plants in general may also be selected from one of the plant families which include Potamogetonaceae, Ceratophyllaceae, Haloragaceae, and Ruppiaceae. More particularly, the aquatic plants chosen should have a large Pasteur effect which increases the ratio of anaerobic CO2 production to the aerobic CO2 production. Typically this ratio is on the order of 1:3, but aquatic plants such as for example Potamogeton pectinatus, commonly known as Sago Pondweed, may increase this ratio to 2:1 as explained in “Anoxia tolerance in the aquatic monocot Potamogeton pectinatus: absence of oxygen stimulates elongation in association with an usually large Pasteur effect,” Journal of Experimental Botany, Volume 51, Number 349, pp. 1413-1422, August 2000, which is incorporated herein by reference. During an elongation process which takes place in an anoxic environment, the plant elongates to form cellular chambers which will later be used to store carbohydrates formed during aerobic metabolism through an aerobic process of the aquatic plant. These carbohydrates can then be used to release ethanol during anaerobic metabolism through an anaerobic process of the aquatic plant. Generally, the equations are as follows:
Aerobic plant metabolism: 6CO2+6H2O→6O2+C6H12O6 
Anaerobic plant metabolism: C6H12O6→2CO2+2C2H5OH
Once the aquatic plants are in a cell, the water in the cell is placed in an anoxic condition by introducing, originally or at a later time, anoxic water into the cell. Alternatively, corn and/or bacteria may be added to the water to deplete the oxygen in the water. The lack of oxygen in the water initiates the anaerobic process in the aquatic plants causing them to elongate and to produce ethanol. This may be encouraged by the introduction of chemical catalysts and CO2. One chemical catalyst which may be included is 2,4-dichlorophenoxyacetic acid. Additional nutrients and salts such as salts of potassium, nitrogen and phosphorus may further be added to promote growth of the aquatic plants. Further, depending upon the species of aquatic plant being utilized, organic substrates, including but not limited to those such as sucrose, glucose and acetate, may also be added to the cell.
During the anaerobic process, the aquatic plants will increase in size dramatically and may achieve a lengthening of up to 10 times or more of its original length. The term ‘size’ here is to be understood to include a volume increase of plant matter which allows for it to store a larger amount of carbohydrates. This elongation provides cellular chambers for holding carbohydrates to be later formed by the aquatic plants. Additionally during the anaerobic process, ethanol is excreted extracellularly by the aquatic plants. This ethanol is then held within the water of the cell until it is removed by conventional methods. As explained further below, the cell, when first used, may be allowed to achieve a minimum ethanol concentration which will be determined depending on the particular method being practiced. This minimum concentration will be increased through progression of the method. This step may take place from one to several days though in the case of Potamogeton pectinatus a total of six days may suffice. The time required will depend on many factors such as light diffusion and availability of nutrients.
The next step is to stop the anaerobic process by creating an oxygenated condition within the cell which will initiate the aerobic process. This may be accomplished by introducing oxygenated water into the cell and by removing the anoxic water. During the aerobic process, as indicated above, the aquatic plants create carbohydrates through metabolic processes and then retain the carbohydrates within their elongated structures. Waste materials, such as waste biomass from the method 10, industrial waste, municipal waste and the like may be added to the cell to provide nutrients to the aquatic plants. Additionally, maximum sunlight filtration is encouraged as is temperature regulation to promote growth of the aquatic plants. Further, the pH of the cell must be monitored to prevent CO2 acidosis of the cell. This may be counteracted with calcium buffering compounds such as calcium carbonate and calcium chlorate, but will ultimately be dependent upon the tolerances of the particular aquatic plant species in the cell. The duration of the aerobic process is likewise dependent upon a number of factors but will typically end when carbohydrate production begins to slow. With Potamogeton pectinatus, this may be between 2 days and 14 days depending upon environmental conditions within the cell.
The use of anoxic and oxygenated water may also be combined by use of thermal strata within the cell. In particular, the coldest strata, which will be on the bottom of the cell, may remain anoxic to encourage growth while the upper strata of warmer water may include oxygenated water to encourage the aerobic process.
Once maximum carbohydrate formation is approached the oxygenated water is removed and is replaced again with anoxic water to again begin the process of elongation and ethanol formation. The steps of adding anoxic water and oxygenated water are then repeated to continually promote elongation and ethanol production followed by carbohydrate production. This creates a self-sustaining cycle as the plant growth replenishes both plant matter lost to plant senescence and those plants which no longer meet established tolerances of ethanol production. Additional plant growth which cannot be used for replenishing purposes or for furnishing plant material for another cell may be removed and fermented using conventional methods to also produce ethanol. Carbon dioxide released during the fermentation process may be captured and returned to the cell to promote carbohydrate production. Plant waste, both before or after the fermentation process, may further be used for replenishing nutrients to the cell and/or may be processed for biochemical industrial usage such as in ethanol and diesel biofuels, pharmaceuticals, cosmetics, colorants, paints and the like.
As stated above, the anoxic water may be retained and used again, at least until its ethanol content approaches a lethal concentration to the aquatic plant. This concentration is dependent upon the aquatic plant being used as well the number of cells being utilized which can affect the number of times the anaerobic process can occur. Typically the method will be practiced with multiple cells wherein the anoxic water and the oxygenated water are rotated between the cells as needed to alternate between the anoxic condition and the oxygenated condition. For example, the process of utilizing multiple cells may include a first cell having anoxic water containing 2% ethanol which is moved into a second cell having previously been oxygenated. The anoxic water replaces the removed oxygenated water in the second cell to create an anoxic condition in the second cell. Within the second cell plant growth and ethanol production are then stimulated. It is noted that having ethanol originally in the second cell (since the anoxic water contains ethanol from the anaerobic process of the first cell) may further spur ethanol production when the aquatic plants detect ethanol in the water. The ethanol concentration may be allowed to increase, for example, up to 4% in the second cell. Each time the anoxic water is moved into a new cell, the elongation and ethanol production of those plants is stimulated. Once the ethanol concentration of the anoxic water reaches a predetermined level, such as for example 10% by volume, the anoxic water is removed from the cell it is now positioned in and the ethanol extracted from the water using conventional means.
The use of multiple cells allows the cycle to be used within a closed loop which again is self sustaining and will sequester carbon dioxide during the formation of carbohydrates. The method 10 grows new aquatic plants faster than they are depleted by senescing to allow for new cells to be seeded by the newly grown aquatic plants. More importantly, all plant waste may be utilized through fermentation into ethanol and processed for biochemical industries or returned to the cells as feed material.
With respect to the above description then, it is to be realized that the optimum dimensional relationships for the parts of an embodiment enabled by the disclosure, to include variations in size, materials, shape, form, function and manner of operation, assembly and use, are deemed readily apparent and obvious to one skilled in the art, and all equivalent relationships to those illustrated in the drawings and described in the specification are intended to be encompassed by an embodiment of the disclosure.
Therefore, the foregoing is considered as illustrative only of the principles of the disclosure. Further, since numerous modifications and changes will readily occur to those skilled in the art, it is not desired to limit the disclosure to the exact construction and operation shown and described, and accordingly, all suitable modifications and equivalents may be resorted to, falling within the scope of the disclosure.

## Claims

1. A method of recovering ethanol from an aquatic plant, said method comprising the steps of:
placing at least on aquatic plant selected from the family Potamogetonaceae in a container containing water,
creating and maintaining an anoxic condition within said water to initiate and maintain anaerobic plant metabolism while inhibiting aerobic plant metabolism therein to produce ethanol, said aquatic plants releasing ethanol into the water during said anoxic condition,
creating and maintaining and oxygenated condition within said water to initiate and maintain aerobic plant metabolism to produce and store carbohydrates while inhibiting anaerobic plant metabolism during said oxygenated condition,
repeating the steps of creating and maintaining anoxic and oxygenated conditions one or more times to repeatedly release ethanol into the water,
recovering ethanol from said water.

2. The method according to claim 1, further including the step of introducing catalysts to said container prior to the step of creating and maintaining an anoxic condition.

3. The method of claim 1, wherein the step of introducing catalysts includes the step of adding 2,4-dichlorophenoxyacetic acid.

4. The method of claim 1, further including the step of adding CO2 to said container prior to the step of creating and maintaining an oxygenated condition.

5. The method of claim 1, further including the steps of:
placing the at least on aquatic plant in at least one additional container; and
transferring said anoxic water between the containers to increase a concentration of ethanol in said anoxic water.

6. The method of claim 1, further including the step of introducing plant nutrients into said container to increase creation of carbohydrates during the step of creating and maintaining an oxygenated condition.