Process for obtaining protein-rich nutrient supplements from bacterial fermentation process
Protein-rich nutrient supplements and animal feed supplements derived from an anaerobic bacterial process are generated through a myriad of cell rupturing and protein fractionation/purification processes. Bacterial fermentation systems and methods of obtaining one or more protein-containing portions from a fermentation process using carbon monoxide-containing gaseous substrates are provided. The invention further provides compositions of protein-rich nutrient supplements with useful applications for intake by a variety of different animals and humans.
1 . A method of producing a protein-rich nutrient supplement having a protein content of 40% to 95% from a bacterial fermentation process, comprising:
fermenting a gaseous substrate with an anaerobic acetogenic bacteria in a fermentation vessel containing a culture medium;
obtaining from the fermentation vessel an amount of first and second fermentation liquid broths;
separating the first fermentation liquid broth in a first cell separator to provide a first cell-free permeate solution and a first cell-containing suspension, wherein the first cell-containing suspension is returned to the fermentation vessel;
separating the second fermentation liquid broth in a second cell separator to provide a second cell-free permeate solution and a second cell-containing suspension, wherein the second cell-containing suspension has a cell concentration of 20 g/L or more;
providing the first cell-free permeate solution and the second cell-free permeate solution to a processing chamber to produce an oxygenated hydrocarbonaceous compound;
adjusting a pH of the second cell-containing suspension to a pH of 7.5 to 10;
rupturing cell membranes of the second cell-containing suspension in one or more rupturing devices to generate a homogenate;
fractionating the homogenate into a first protein-containing portion and a protein-containing cell debris portion using a first fractionator; and
obtaining the first protein-containing portion, wherein the protein-rich nutrient supplement is processed and produced from a portion selected from the group consisting of the first protein-containing portion, the protein-containing debris portion, and combinations thereof.
2 . The method of claim 1 , wherein the first protein-containing portion has a protein content that is between 10% and 95%.
3 . The method of claim 1 , wherein the first and second fermentation liquid broths containing the anaerobic bacterial cells are separated into the first and second cell-free permeate solutions and the first and second cell-containing suspensions by ultrafiltration.
4 . The method of claim 1 , wherein the first cell-free permeate solution comprises one or more fermentation product selected from the group consisting of ethanol, butanol, acetic acid, butyric acid, and combinations thereof.
5 . The method of claim 1 , further comprising:
treating the second cell-containing suspension with one or more additives prior to the rupturing of cell membranes of the second cell-containing, wherein the one or more additives are selected from the group consisting of surfactants, detergents, EDTA, sodium dodecyl sulfate, CHAPS, enzyme, proteases, lysozymes, benzonases, nucleases, ribonucleases (RNases), deoxyribonucleases (DNases), hydrolysis-inducing agents, pH-adjusting agents, and combinations thereof.
6 . The method of claim 1 , further comprising:
treating the second cell-containing suspension with one or more additives prior to the fractionating the first protein-containing portion from the cell debris portion, wherein the one or more additives are selected from the group consisting of surfactants, detergents, EDTA, sodium dodecyl sulfate, CHAPS, enzyme, proteases, lysozymes, benzonases, nucleases, ribonucleases (RNases), deoxyribonucleases (DNases), hydrolysis-inducing agents, pH-adjusting agents, and combinations thereof.
7 . The method of claim 1 , wherein the first fractionator is selected from the group consisting of a solid-liquid fractionator, a centrifugation device, a continuous centrifuge, a decanter centrifuge, a disc-stack centrifuge, a filtration device, a hollow fiber filtration device, a spiral wound filtration device, a ceramic filter device, a cross-flow filtration device, a size exclusion device, one or series of size exclusion columns, one or series of ion exchange columns, one or series of carbon polymer columns, a flow-through magnetic fractionator, an ultrafiltration device, one or series of affinity chromatography columns, one or series of gel filtration columns, and combinations thereof.
8 . The method of claim 1 , where the one or more rupturing devices are selected from the group consisting of a microfluidic device, a sonication device, an ultrasonic device, a French press, and combinations thereof.
9 . A method of producing a protein-rich nutrient supplement having a protein content of 40% to 95% from a bacterial fermentation process, comprising:
fermenting a gaseous substrate with anaerobic acetogenic bacteria in a fermentation vessel;
obtaining from the fermentation vessel an amount of a first and second fermentation liquid broths;
separating the first fermentation liquid broth in a first cell separator to provide a first cell-free permeate solution and a first cell-containing suspension, wherein the first cell-containing suspension is returned to the fermentation vessel;
separating the second fermentation liquid broth in a second cell separator to provide a second cell-free permeate solution and a second cell-containing suspension, wherein the second cell-containing suspension has a cell concentration of 20 g/L or more;
providing the first cell-free permeate solution and the second cell-free permeate solution to a processing chamber to produce an oxygenated hydrocarbonaceous compound;
holding the second cell-containing suspension in a cell-containing holding tank;
delivering the second cell-containing suspension from the cell-containing holding tank at a delivery rate to one or more rupturing devices, wherein a pH of the second cell-containing suspension from the cell-containing holding tank is adjusted to a pH of 7.5 to 10 prior to rupturing;
rupturing cell membranes of the second cell containing suspension using the in one or more rupturing devices to generate a homogenate; and
fractionating the homogenate into a first protein-containing portion and a protein-containing cell debris portion using one or more fractionators, wherein the protein-rich nutrient supplement is processed and produced from a portion selected from the group consisting of the first protein-containing portion, the protein-containing debris portion, and combinations thereof.
10 . The method of claim 9 , wherein the cell-containing holding tank is a pretreatment chamber to house the second cell-containing suspension such that the second cell-containing suspension undergoes a pretreatment process, wherein the pretreatment process comprises treating the second cell-containing suspension with one or more additives, wherein the one or more additives are selected from the group consisting of surfactants, detergents, EDTA, sodium dodecyl sulfate, CHAPS, enzyme, proteases, lysozymes, benzonases, nucleases, ribonucleases (RNases), deoxyribonucleases (DNases), hydrolysis-inducing agents, pH-adjusting agents, and combinations thereof.
11 . The method of claim 9 , where the one or more rupturing devices are selected from the group consisting of a microfluidic device, a sonication device, an ultrasonic device, a French press, and combinations thereof.
12 . A method of producing a protein-rich nutrient supplement having a protein content of 40% to 95% from a bacterial fermentation process, comprising:
fermenting a gaseous substrate with an anaerobic acetogenic bacteria in a fermentation vessel containing a culture medium;
obtaining from the fermentation vessel an amount of a first and second fermentation liquid broths;
separating the first fermentation liquid broth in a first cell separator to provide a first cell-free permeate solution and a first cell-containing suspension, wherein the first cell-containing suspension is returned to the fermentation vessel;
separating the second fermentation liquid broth in a second cell separator to provide a second cell-free permeate solution and a second cell-containing suspension, wherein the second cell-containing suspension has a cell concentration of 20 g/L or more;
providing the first cell-free permeate solution and the second cell-free permeate solution to a processing chamber to produce an oxygenated hydrocarbonaceous compound;
adjusting a pH of the second cell-containing suspension to a pH of 7.5 to 10;
rupturing cell membranes of the second cell-containing suspension in one or more rupturing devices to generate a homogenate;
fractionating the homogenate into a first protein-containing portion and a protein-containing cell debris portion using a first fractionator;
obtaining the first protein-containing portion;
delivering the first protein-containing portion to a second fractionator;
fractionating the first protein-containing portion into a second protein-containing portion using the second fractionator; and
collecting the second protein-containing portion,
wherein the second protein-containing portion is produced as the protein-rich nutrient supplement.
13 . The method of claim 12 , wherein the second fractionator is selected from the group consisting of a solid-liquid fractionator, a centrifugation device, a continuous centrifuge, a decanter centrifuge, a disc-stack centrifuge, a filtration device, a hollow fiber filtration device, a spiral wound filtration device, a ceramic filter device, a cross-flow filtration device, a size exclusion device, one or series of size exclusion columns, one or series of ion exchange columns, one or series of carbon polymer columns, a flow-through magnetic fractionator, an ultrafiltration device, one or series of affinity chromatography columns, one or series of gel filtration columns, and combinations thereof.
14 . The method of claim 12 , where the one or more rupturing devices are selected from the group consisting of a microfluidic device, a sonication device, an ultrasonic device, a French press, and combinations thereof.
15 . A method of producing a protein-rich nutrient supplement having a protein content of 40% to 95% from a bacterial fermentation process, comprising:
fermenting a gaseous substrate with an anaerobic acetogenic bacteria in a fermentation vessel containing a culture medium;
obtaining from the fermentation vessel an amount of a first and second fermentation liquid broths;
separating the first fermentation liquid broth in a first cell separator to provide a first cell-free permeate solution and a first cell-containing suspension, wherein the first cell-containing suspension is returned to the fermentation vessel;
separating the second fermentation liquid broth in a second cell separator to provide a second cell-free permeate solution and a second cell-containing suspension, wherein the second cell-containing suspension has a cell concentration of 20 g/L or more;
providing the first cell-free permeate solution and the second cell-free permeate solution to a processing chamber to produce an oxygenated hydrocarbonaceous compound;
adjusting a pH of the second cell-containing suspension to a pH of 7.5 to 10;
rupturing cell membranes of the second cell-containing suspension in one or more rupturing devices to generate a homogenate;
fractionating the homogenate into a first protein-containing portion and a protein-containing cell debris portion using a first fractionator;
obtaining the first protein-containing portion;
delivering the first protein-containing portion to a second fractionator;
fractionating the first protein-containing portion into a second protein-containing portion using the second fractionator;
collecting the second protein-containing portion;
delivering the second protein-containing portion to a third fractionator;
fractionating the second protein-containing portion into a third protein-containing portion using the third fractionator; and
collecting the third protein-containing portion,
wherein the third protein-containing portion is produced as the protein-rich nutrient supplement.
16 . The method of claim 15 , wherein the third fractionator is selected from the group consisting of a solid-liquid fractionator, a centrifugation device, a continuous centrifuge, a decanter centrifuge, a disc-stack centrifuge, a filtration device, a hollow fiber filtration device, a spiral wound filtration device, a ceramic filter device, a cross-flow filtration device, a size exclusion device, one or series of size exclusion columns, one or series of ion exchange columns, one or series of carbon polymer columns, a flow-through magnetic fractionator, an ultrafiltration device, one or series of affinity chromatography columns, one or series of gel filtration columns, and combinations thereof.
17 . The method of claim 15 , where the one or more rupturing devices are selected from the group consisting of a microfluidic device, a sonication device, an ultrasonic device, a French press, and combinations thereof.