Production of biofuel from tissue culture sources

Abstract

The present invention provides a method for the production of at least one fatty acid and/or oil from a plant cell suspension culture, the method comprising (i) maintaining a cell suspension culture of oil-producing plant cells under conditions such that the cultured cells synthesise and secrete at least one fatty acid and/or oil into the cell suspension culture medium; and (ii) extracting the thus secreted at least one fatty acid and/or oil from the cell suspension culture medium.

1 - 118 . (canceled) 
     
     
         119 . A method for the production of at least one fatty acid and/or oil from a plant cell suspension culture, the method comprising:
 (i) maintaining a cell suspension culture of oil-producing plant cells under conditions such that the cultured cells synthesise and secrete at least one fatty acid and/or oil into the cell suspension culture medium; and   (ii) extracting the thus secreted at least one fatty acid and/or oil from the cell suspension culture medium.   
     
     
         120 . The method of  claim 119 , wherein the plant cell suspension culture is maintained at a pH suitable to cause fatty acids and/or oils stored in the vacuole of the cultured plant cells to be released, via the cytosol of the plant cells, into the cell suspension culture medium. 
     
     
         121 . The method of  claim 120 , wherein the cell suspension culture medium comprises a buffer that maintains the cell suspension culture medium at around the selected pH. 
     
     
         122 . The method of  claim 119 , wherein the viability of the plant cell culture is maintained during the step of extracting the thus secreted at least one fatty acid and/or oil from the cell suspension culture medium. 
     
     
         123 . The method of  claim 119 , wherein the method is a method for the continuous harvest of at least one fatty acid and/or oil from the cell suspension culture medium. 
     
     
         124 . The method of  claim 119 , wherein the pH at which the cell suspension culture medium is maintained is selected to promote the release of the secreted fatty acids and/or oils from an emulsion within the cell suspension culture medium. 
     
     
         125 . The method of  claim 124 , wherein the secretion of the at least one fatty acid and/or oil from the cultured cells into the surrounding cell suspension culture medium results in the formation of a biphasic system in which the at least one fatty acid and/or oil collects in a separate layer to the cell suspension culture medium. 
     
     
         126 . The method of  claim 125 , wherein the step of extracting the thus secreted at least one fatty acid and/or oil from the cell suspension culture medium comprises direct extraction of the at least one fatty acid and/or oil from the layer that it forms within the biphasic system. 
     
     
         127 . The method of  claim 119 , wherein the oil-producing plant cell is a differentiated plant cell. 
     
     
         128 . The method of  claim 127 , wherein the differentiated plant cell is a cell that is specialised in the production and storage of oils. 
     
     
         129 . The method of  claim 119 , wherein the oil-producing plant cell is from an oil-producing plant. 
     
     
         130 . The method of  claims 119 , wherein the at least one fatty acid and/or oil that is extracted is then further processed to convert it to a biofuel, or is optionally further purified and/or used in a downstream process such as by incorporation into a food product, cosmetic, or lubricant. 
     
     
         131 . A cell suspension culture of oil-producing plant cells as defined by  claim 119 , wherein the pH of the cell suspension culture is less than about pH 5.5. 
     
     
         132 . A buffered plant cell suspension culture medium having a pH of less than about pH 5.5 for culturing a plant cell suspension culture by a method according to  claim 119 . 
     
     
         133 . A product selected from the group consisting of an extract of at least one fatty acid and/or oil obtainable by the method of  claim 119 , and a product comprising the extract. 
     
     
         134 . A biofuel obtainable by the method of  claim 130 . 
     
     
         135 . A method for the production of a biological product, the method comprising:
 (i) maintaining a first cell suspension culture of photosynthetic plant cells under conditions that allow the cultured cells to photosynthesise and thereby generate and release sugars into the surrounding culture medium; and   (ii) maintaining a second cell culture in the presence of the sugars generated by the first cell suspension culture to allow growth of the second culture and the production of a biological product.   
     
     
         136 . The method of  claim 135 , wherein the biological product is the cells of the second cell culture. 
     
     
         137 . The method of  claim 135 , wherein the biological product is synthesised by the cells of the second cell culture. 
     
     
         138 . The method of  claim 135 , wherein the cells of the first cell suspension culture and the cells of the second cell culture are in fluid communication with each other. 
     
     
         139 . The method of  claim 138 , wherein the cells of the first cell suspension culture and the cells of the second cell culture are mixed together and cultured in the same medium. 
     
     
         140 . The method of  claim 138 , wherein cells of the first cell suspension culture and the cells of the second suspension culture are cultured in separate culture vessels that are connected in fluid communication. 
     
     
         141 . The method of  claim 138 , wherein the fluid communication between the cells of the first cell suspension culture and the cells of the second cell culture allows the sugar released by the cells of the first cell suspension culture to be used as a carbon source by the cells of the second cell culture. 
     
     
         142 . The method of  claim 138 , wherein the cells of the first cell suspension culture and the cells of the second cell culture are each grown in separate culture vessels that are not in fluid communication with each other. 
     
     
         143 . The method of  claim 142 , that comprises the step of extracting sugar from the culture medium of the first cell suspension culture and the further step of feeding the extracted sugar into the second cell culture. 
     
     
         144 . The method of  claim 143 , wherein the step of extracting sugar from the culture medium of the first cell suspension culture comprises separating the sugar-containing medium of the first cell culture from the cells present therein. 
     
     
         145 . The method of  claim 143 , wherein the step of extracting sugar from the culture medium of the first cell suspension culture comprises continuously removing sugar from the cell culture medium of the first cell culture. 
     
     
         146 . The method of  claim 135 , wherein the second cell culture is maintained in the presence of sugar generated by the first cell suspension culture at a sugar concentration in the range of 20 g/L to 400 g/L. 
     
     
         147 . The method of  claim 137 , wherein the biological product is selected from the group consisting of at least one fatty acid and/or oil, a proteinaceous product and a metabolite. 
     
     
         148 . The method of  claim 135 , wherein the cells of the second cell culture are selected from the group consisting of prokaryotic cells, eukaryotic cells, bacterial cells, fungal cells, plant cells, animal cells and human cells. 
     
     
         149 . The method of  claim 135 , wherein the cells of the second cell culture are microorganisms and optionally the biological product is an alcohol. 
     
     
         150 . The method of  claim 135 , which is a method for the production of at least one fatty acid and/or oil from a plant cell culture, the method comprising maintaining a second cell suspension culture of oil-producing plant cells in the presence of the sugar generated by the first cell suspension culture and under conditions such that the cultured oil-producing plant cells produce at least one fatty acid and/or oil. 
     
     
         151 . The method of  claim 135 , further comprising the step of extracting the biological product from the second cell culture. 
     
     
         152 . The method of  claim 150 , further comprising the step of extracting the at least one fatty acid and/or oil from the second cell suspension culture of oil-producing plant cells. 
     
     
         153 . The method of  claim 151 , comprising further purifying and/or processing the thus-extracted biological product. 
     
     
         154 . The method of  claim 152 , comprising further purifying and/or processing the thus-extracted at least one fatty acid and/or oil, such as by further processing it to convert it to a biofuel, and/or further using it in a downstream process such as by incorporation into a food product, cosmetic, or lubricant. 
     
     
         155 . The method of  claim 152 , wherein the steps of maintaining a second cell suspension culture of oil-producing plant cells in the presence of the sugar generated by the first cell suspension culture and under conditions such that the cultured oil-producing plant cells produce at least one fatty acid and/or oil, and extracting the at least one fatty acid and/or oil from the second cell suspension culture of oil-producing plant cells, is performed in accordance with a method comprising:
 (i) maintaining the second cell suspension culture of oil-producing plant cells under conditions such that the cultured cells synthesise and secrete at least one fatty acid and/or oil into the cell suspension culture medium; and   (ii) extracting the thus secreted at least one fatty acid and/or oil from the cell suspension culture medium.   
     
     
         156 . The method of  claim 155 , wherein the at least one fatty acid and/or oil that is extracted is then further processed to convert it to a biofuel, or is optionally further purified and/or used in a downstream process such as by incorporation into a food product, cosmetic, or lubricant. 
     
     
         156 . The method of  claim 135 , wherein the photosynthetic plant cells present in the first cell suspension culture are differentiated photosynthetic plant cells. 
     
     
         157 . The method of  claim 156 , wherein the differentiated plant cell is a cell that is specialised for photosynthesis. 
     
     
         158 . The method of  claim 157 , wherein the photosynthetic plant cells present in the first cell suspension culture are palisade cells. 
     
     
         160 . The method of  claim 135 , wherein the photosynthetic plant cells present in the first cell suspension culture of are isolated from a copper-tolerant plant. 
     
     
         161 . The method of  claim 135 , wherein the first cell suspension culture of photosynthetic plant cells has a cell culture medium copper level of up to 0.1M. 
     
     
         162 . The method of  claim 150 , wherein the oil-producing plant cell present in the second cell suspension culture is a differentiated plant cell. 
     
     
         163 . The method of  claim 162 , wherein the differentiated plant cell is a cell that is specialised in the production and storage of oils.) 
     
     
         164 . The method of  claim 135 , wherein the first cell suspension culture of photosynthetic plant cells is fed carbon dioxide from a carbon dioxide source selected from the group consisting of liquid carbon dioxide and gaseous carbon dioxide. 
     
     
         165 . The method of  claim 164 , wherein the carbon dioxide source is obtained as a by-product of a carbon dioxide-producing process. 
     
     
         166 . The method of  claim 165  wherein at least the first cell suspension culture, and optionally also the second cell culture, is or are maintained at the site of the carbon dioxide-producing process. 
     
     
         167 . A two-culture system for producing a biological product, comprising a first plant cell suspension culture and a second cell culture, each as defined by  claim 135 . 
     
     
         168 . The two-culture system of  claim 167 , that further comprises a carbon dioxide-generating source, and wherein the thus-generated carbon dioxide is fed into the first plant cell suspension culture. 
     
     
         169 . The two-culture system of  claim 168 , for producing at least one fatty acid and/or oil, comprising a first plant cell suspension culture of photosynthetic plant cells that are maintained under conditions that allow the cultured cells to photosynthesise and thereby generate and release sugars into the surrounding culture medium, and a second plant cell suspension culture of oil-producing plant cells that are maintained in the presence of the sugar generated by the first cell suspension culture under conditions that allow growth of the second culture and allow the second culture to produce at least one fatty acid and/or oil. 
     
     
         170 . A carbon dioxide capture system comprising at least a first plant cell suspension culture of photosynthetic plant cells that are maintained under conditions that allow the cultured cells to photosynthesise and thereby generate and release sugars into the surrounding culture medium, and optionally also a second cell culture that is maintained in the presence of the sugar generated by the first cell suspension culture to allow growth of the second culture and the production of a biological product. 
     
     
         171 . The carbon dioxide capture system of  claim 170 , that comprises a carbon dioxide-generating source, and wherein the thus-generated carbon dioxide is fed into the first plant cell suspension culture. 
     
     
         172 . The carbon dioxide capture system of  claim 170 , which comprises a second plant cell suspension culture of oil-producing plant cells that are maintained in the presence of the sugar generated by the first cell suspension culture under conditions that allow growth of the second culture and allow the second culture to produce at least one fatty acid and/or oil. 
     
     
         173 . A carbon dioxide-producing power or biofuel generating facility comprising the two-culture system of  claim 167 . 
     
     
         174 . A carbon dioxide-producing power or biofuel generating facility comprising the carbon dioxide capture system of  claim 170 . 
     
     
         175 . The carbon dioxide-producing power generating facility of  claim 173 , wherein the two-culture system produces at least one fatty acid and/or oil from the captured carbon dioxide and the thus produced at least one fatty acid and/or oil is used to supplement the fuel used by the power generating facility. 
     
     
         176 . The carbon dioxide-producing power generating facility of  claim 175 , wherein the carbon dioxide capture system produces at least one fatty acid and/or oil from the captured carbon dioxide and the thus produced at least one fatty acid and/or oil is used to supplement the fuel used by the power generating facility. 
     
     
         177 . The carbon dioxide-producing biofuel generating facility of  claim 173 , wherein sugars produced by the first cell suspension culture of photosynthetic plant cells present within the two-culture system are used to supplement the growth of microorganisms used in the production of biofuel by the biofuel generating facility. 
     
     
         178 . The carbon dioxide-producing biofuel generating facility of  claim 174 , wherein sugars produced by the first cell suspension culture of photosynthetic plant cells present within the carbon dioxide capture system are used to supplement the growth of microorganisms used in the production of biofuel by the biofuel generating facility. 
     
     
         179 . An extract of a biological product obtainable by the method of  claim 151 . 
     
     
         180 . An extract of at least one fatty acid and/or oil obtainable by the method of  claim 152 . 
     
     
         181 . A product selected from the group consisting of a biofuel, food product, cosmetic or lubricant, wherein the product is obtainable by method of  claim 154 . 
     
     
         182 . A further purified or processed product obtainable by the method of  claim 153 . 
     
     
         183 . An extract of at least one fatty acid and/or oil obtainable by the method of  claim 155 . 
     
     
         184 . A product selected from the group consisting of a biofuel, food product, cosmetic or lubricant, wherein the product is obtainable by the method of  claim 156 .