Methods and apparatus for enzymatically producing ethanol

Claim 1

A method of producing ethanol comprising providing a first reaction zone containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, providing a second reaction zone containing the enzymatic proteins aldose, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase, providing a third reaction zone containing the enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase, providing a fourth reaction zone containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, retaining said enzymatic proteins in their respective reaction zones, passing a fermentable sugar solution successively through said reaction zones, adding coenzymes and cofactors to said reaction zones as required for the enzymatic proteins in each zone to perform their catalytic functions, and recovering ethanol from the fourth reaction zone.

Claim 2

A method of producing ethanol comprising providing a first reaction zone containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, providing a second reaction zone containing the enzymatic proteins aldose, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase, providing a third reaction zone containing the enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase, providing a fourth reaction zone containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, retaining said enzymatic proteins in their respective reaction zones, passing a fermentable sugar solution successively through said reaction zones, adding coenzymes and cofactors to said reaction zones as required for the enzymatic proteins in each zone to perform their catalytic functions, and recovering ethanol from the fourth reaction zone.

Claim 7

A method of producing ethanol comprising immobilizing the enzymatic proteins whose combined catalytic activity completes the reaction sequence of the glycolytic pathway for the conversion of fermentable sugars to ethanol, disposing said immobilized enzymatic proteins in four segregated reaction chambers arranged in accordance with the reaction sequence of the glycolytic pathway, adding coenzymes and cofactors to a fermentable sugar substrate solution, introducing said sugar substrate solution plus said additives into said first chamber containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, introducing the solution exiting from said first chamber into said second chamber containing the enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase, introducing the solution exiting from said second chamber into said third chamber containing the enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase, introducing the solution exiting from said third chamber into said fourth chamber containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, and recovering ethanol from said fourth chamber.

Claim 8

A method of producing ethanol comprising immobilizing the enzymatic proteins whose combined catalytic activity completes the reaction sequence of the glycolytic pathway for the conversion of fermentable sugars to ethanol, disposing said immobilized enzymatic proteins in four segregated reaction chambers arranged in accordance with the reaction sequence of the glycolytic pathway, adding coenzymes and cofactors to a fermentable sugar substrate solution, introducing said sugar substrate solution plus said additives into said first chamber containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, introducing the solution exiting from said first chamber into said second chamber containing the enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase, introducing the solution exiting from said second chamber into said third chamber containing the enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase, introducing the solution exiting from said third chamber into said fourth chamber containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, and recovering ethanol from said fourth chamber.

Claim 13

A system for enzymatically convering sugar to ethanol comprising four successive reaction chambers containing enzymatic proteins arranged in sequence for conversion of sugar to ethanol and CO 2 by the glycolytic pathway, said first chamber containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, said second chamber containing the enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase, said third chamber containing the enzymatic proteins phosphoglyceromutase, enolase, pyruvate kinase, said fourth chamber containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, means for retaining said enzymatic proteins in each of said chambers, means for causing a fermentable sugar substrate solution to flow sequentially through said reaction chambers so as to come in contact with said enzymatic proteins, means for introducing necessary coenzymes and cofactors to said reaction chambers along with said flowing sugar substrate solution, and means for recovering ethanol from said fourth reaction chamber.

Claim 14

A system for enzymatically convering sugar to ethanol comprising four successive reaction chambers containing enzymatic proteins arranged in sequence for conversion of sugar to ethanol and CO 2 by the glycolytic pathway, said first chamber containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase and phosphofructokinase, said second chamber containing the enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase, said third chamber containing the enzymatic proteins phosphoglyceromutase, enolase, pyruvate kinase, said fourth chamber containing the enzymatic proteins pyruvate decarboxylase and alcohol dehydrogenase, means for retaining said enzymatic proteins in each of said chambers, means for causing a fermentable sugar substrate solution to flow sequentially through said reaction chambers so as to come in contact with said enzymatic proteins, means for introducing necessary coenzymes and cofactors to said reaction chambers along with said flowing sugar substrate solution, and means for recovering ethanol from said fourth reaction chamber.

Claim 15

A method of producing ethanol comprising immobilizing the enzymatic proteins whose combined catalytic activity completes the reaction sequence of the glycolytic pathway for the conversions of fermentable sugars to ethanol, said enzymes including an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase, and alcohol dehydrogenase, grouping said immobilized enzymatic proteins in sequenced reaction zones, the enzymatic proteins phosphoglucose isomerase and phosphofructokinase being grouped together in one reaction zone, said enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase being grouped together in another reaction zone and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped in yet another reaction zone, adding coenzymes and cofactors to a fermentable sugar substrate solution, flowing said sugar substrate solution plus said additives to said sequenced reaction zones containing said immobilized enzymatic proteins, and recovering ethanol from the last zone.

Claim 16

A method of producing ethanol comprising immobilizing the enzymatic proteins whose combined catalytic activity completes the reaction sequence of the glycolytic pathway for the conversions of fermentable sugars to ethanol, said enzymes including an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase, and alcohol dehydrogenase, grouping said immobilized enzymatic proteins in sequenced reaction zones, the enzymatic proteins phosphoglucose isomerase and phosphofructokinase being grouped together in one reaction zone, said enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase being grouped together in another reaction zone and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped in yet another reaction zone, adding coenzymes and cofactors to a fermentable sugar substrate solution, flowing said sugar substrate solution plus said additives to said sequenced reaction zones containing said immobilized enzymatic proteins, and recovering ethanol from the last zone.

Claim 18

A method of producing ethanol comprising Providing a sequenced series of reaction zones containing in total the enzymatic proteins which catalyze the steps the glycolytic pathway that converts a fermentable sugar into ethanol, the enzymatic proteins including in sequence an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase, and alcohol dehydrogenase, said enzymatic proteins phosphyglucose isomerase and phosphofructokinase being grouped together in one reaction zone, said enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase being grouped together in another reaction zone and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped together in yet another reaction zone, retaining said enzymatic proteins in their respective reaction zones, passing a fermentable sugar solution successively through said reaction zones, adding coenzymes and cofactors to said reaction zones as required for the enzymatic proteins in each reaction zone to perform their catalytic function, and recovering ethanol from the last reaction zone.

Claim 19

A method of producing ethanol comprising Providing a sequenced series of reaction zones containing in total the enzymatic proteins which catalyze the steps the glycolytic pathway that converts a fermentable sugar into ethanol, the enzymatic proteins including in sequence an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase, and alcohol dehydrogenase, said enzymatic proteins phosphyglucose isomerase and phosphofructokinase being grouped together in one reaction zone, said enzymatic proteins aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase being grouped together in another reaction zone and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped together in yet another reaction zone, retaining said enzymatic proteins in their respective reaction zones, passing a fermentable sugar solution successively through said reaction zones, adding coenzymes and cofactors to said reaction zones as required for the enzymatic proteins in each reaction zone to perform their catalytic function, and recovering ethanol from the last reaction zone.

Claim 23

A system for enzymatically converting sugar to ethanol comprising a plurality of reaction chambers containing enzymatic proteins arranged in sequence for conversion of sugar to ethanol by the glycolytic pathway, said reaction chambers in total containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase and alcohol dehydrogenase, means for retaining said enzymatic proteins in each of said chambers, the enzymatic proteins phosphoglucose isomerase and phosphofructokinase being grouped together in one chamber, the enzymatic proteins aldolase, triose phophate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase being grouped together in another chamber and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped together in another chamber.

Claim 24

A system for enzymatically converting sugar to ethanol comprising a plurality of reaction chambers containing enzymatic proteins arranged in sequence for conversion of sugar to ethanol by the glycolytic pathway, said reaction chambers in total containing an enzymatic protein selected from the group consisting of hexokinase and glucokinase, plus the enzymatic proteins phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, pyruvate kinase, pyruvate decarboxylase and alcohol dehydrogenase, means for retaining said enzymatic proteins in each of said chambers, the enzymatic proteins phosphoglucose isomerase and phosphofructokinase being grouped together in one chamber, the enzymatic proteins aldolase, triose phophate isomerase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase being grouped together in another chamber and said enzymatic proteins phosphoglyceromutase, enolase and pyruvate kinase being grouped together in another chamber.