(3-carboxy-2-oxopropyl)-trimethylazanium;chloride

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine was synthesized as in example 2, except that the NADH regeneration system comprised, in addition to alcohol dehydrogenase, aldehyde dehydrogenase (E.C. 1.2.1.5) which oxidizes the aldehyde (formed by action of alcohol dehydrogenase on ethanol) to acetic acid. Two molecules of NADH were formed from one molecule of ethanol (instead of one in the preceeding example) and the reaction balance was strongly displaced in favour of NADH and acetic acid formation. It was thus no longer necessary to eliminate acetaldehyde as in the preceeding example. The conditions were thus those of example 2, except that 125 units of purified aldehyde dehydrogenase (Boehringer ref. 171.832) were added to the reaction medium and nitrogen bubbling was omitted. After 24 hours, 0.764 g of L-carnitine was formed with a yield of 77.5% with respect to dehydrocarnitine hydrochloride.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine dehydrogenase obtained as in example 1 was used in that synthesis. Synthesis of L-carnitine was carried out in the same conditions as in example 1, except as concerns the NADH regeneration system which was the yeast alcohol dehydrogenase and ethanol system instead of the glucose dehydrogenase and glucose system. Glucose was thus substituted with ethanol (concentration in the reactor: 4% by volume) and glucose dehydrogenase with 4 mg (700 units) of purified yeast alcohol dehydrogenase (Sigma ref. A 3263). Nitrogen was bubbled through the reactor (80 ml/mn) to eliminate the acetaldehyde formed by alcohol oxidation and thus improve the NADH regeneration. After 30 h, 0.325 g of L-carnitine was produced from dehydrocarnitine hydrochloride with a yield of 27%.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: The reaction product was passed through a column of cation exchange resin Dowex AG 50W-X8 of 20×200 mm. This column had been previously brought to the H+ form by washing with 350 ml of 10% HCl and then with 500 ml of water. After introduction of the sample, the column was washed with 400 ml of water and then eluted with a 1-normal ammonia solution. The fraction discharged from the column between 150 ml and 225 ml, after the beginning of the elution with ammonia, was collected. By carnitine acetyltransferase titration, it was found to contain 1.48 g of carnitine. The sample was then evaporated in vacuo, re-dissolved into water (15 ml) and evaporated again. The carnitine base was converted to the hydrochloride by addition of 8 ml of a 1-normal HCl solution and evaporated again. The deliquescent product was dissolved into 75 ml of an anhydrous mixture of 3 parts of acetone with 2 parts of ethanol. 210 ml of acetone at 0° C. was added and the solution was stored at -9° C. for 12 hours. The crystals were collected, washed with ether and dried under vacuum. 0.91 g of carnitine was obtained. The final product was titrated with carnitine acetyltransferase and found to contain 89.5% of L-carnitine hydrochloride. It was obtained with an aggregate yield of 57% with respect to dehydrocarnitine hydrochloride.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine was synthesized as in example 2, except that the NADH regeneration system comprised, in addition to alcohol dehydrogenase, aldehyde dehydrogenase (E.C. 1.2.1.5) which oxidizes the aldehyde (formed by action of alcohol dehydrogenase on ethanol) to acetic acid. Two molecules of NADH were formed from one molecule of ethanol (instead of one in the preceeding example) and the reaction balance was strongly displaced in favour of NADH and acetic acid formation. It was thus no longer necessary to eliminate acetaldehyde as in the preceeding example. The conditions were thus those of example 2, except that 125 units of purified aldehyde dehydrogenase (Boehringer ref. 171.832) were added to the reaction medium and nitrogen bubbling was omitted. After 24 hours, 0.764 g of L-carnitine was formed with a yield of 77.5% with respect to dehydrocarnitine hydrochloride.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine dehydrogenase obtained as in example 1 was used in that synthesis. Synthesis of L-carnitine was carried out in the same conditions as in example 1, except as concerns the NADH regeneration system which was the yeast alcohol dehydrogenase and ethanol system instead of the glucose dehydrogenase and glucose system. Glucose was thus substituted with ethanol (concentration in the reactor: 4% by volume) and glucose dehydrogenase with 4 mg (700 units) of purified yeast alcohol dehydrogenase (Sigma ref. A 3263). Nitrogen was bubbled through the reactor (80 ml/mn) to eliminate the acetaldehyde formed by alcohol oxidation and thus improve the NADH regeneration. After 30 h, 0.325 g of L-carnitine was produced from dehydrocarnitine hydrochloride with a yield of 27%.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: The reaction product was passed through a column of cation exchange resin Dowex AG 50W-X8 of 20×200 mm. This column had been previously brought to the H+ form by washing with 350 ml of 10% HCl and then with 500 ml of water. After introduction of the sample, the column was washed with 400 ml of water and then eluted with a 1-normal ammonia solution. The fraction discharged from the column between 150 ml and 225 ml, after the beginning of the elution with ammonia, was collected. By carnitine acetyltransferase titration, it was found to contain 1.48 g of carnitine. The sample was then evaporated in vacuo, re-dissolved into water (15 ml) and evaporated again. The carnitine base was converted to the hydrochloride by addition of 8 ml of a 1-normal HCl solution and evaporated again. The deliquescent product was dissolved into 75 ml of an anhydrous mixture of 3 parts of acetone with 2 parts of ethanol. 210 ml of acetone at 0° C. was added and the solution was stored at -9° C. for 12 hours. The crystals were collected, washed with ether and dried under vacuum. 0.91 g of carnitine was obtained. The final product was titrated with carnitine acetyltransferase and found to contain 89.5% of L-carnitine hydrochloride. It was obtained with an aggregate yield of 57% with respect to dehydrocarnitine hydrochloride.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine was synthesized as in example 2, except that the NADH regeneration system comprised, in addition to alcohol dehydrogenase, aldehyde dehydrogenase (E.C. 1.2.1.5) which oxidizes the aldehyde (formed by action of alcohol dehydrogenase on ethanol) to acetic acid. Two molecules of NADH were formed from one molecule of ethanol (instead of one in the preceeding example) and the reaction balance was strongly displaced in favour of NADH and acetic acid formation. It was thus no longer necessary to eliminate acetaldehyde as in the preceeding example. The conditions were thus those of example 2, except that 125 units of purified aldehyde dehydrogenase (Boehringer ref. 171.832) were added to the reaction medium and nitrogen bubbling was omitted. After 24 hours, 0.764 g of L-carnitine was formed with a yield of 77.5% with respect to dehydrocarnitine hydrochloride.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Carnitine dehydrogenase obtained as in example 1 was used in that synthesis. Synthesis of L-carnitine was carried out in the same conditions as in example 1, except as concerns the NADH regeneration system which was the yeast alcohol dehydrogenase and ethanol system instead of the glucose dehydrogenase and glucose system. Glucose was thus substituted with ethanol (concentration in the reactor: 4% by volume) and glucose dehydrogenase with 4 mg (700 units) of purified yeast alcohol dehydrogenase (Sigma ref. A 3263). Nitrogen was bubbled through the reactor (80 ml/mn) to eliminate the acetaldehyde formed by alcohol oxidation and thus improve the NADH regeneration. After 30 h, 0.325 g of L-carnitine was produced from dehydrocarnitine hydrochloride with a yield of 27%.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: The reaction product was passed through a column of cation exchange resin Dowex AG 50W-X8 of 20×200 mm. This column had been previously brought to the H+ form by washing with 350 ml of 10% HCl and then with 500 ml of water. After introduction of the sample, the column was washed with 400 ml of water and then eluted with a 1-normal ammonia solution. The fraction discharged from the column between 150 ml and 225 ml, after the beginning of the elution with ammonia, was collected. By carnitine acetyltransferase titration, it was found to contain 1.48 g of carnitine. The sample was then evaporated in vacuo, re-dissolved into water (15 ml) and evaporated again. The carnitine base was converted to the hydrochloride by addition of 8 ml of a 1-normal HCl solution and evaporated again. The deliquescent product was dissolved into 75 ml of an anhydrous mixture of 3 parts of acetone with 2 parts of ethanol. 210 ml of acetone at 0° C. was added and the solution was stored at -9° C. for 12 hours. The crystals were collected, washed with ether and dried under vacuum. 0.91 g of carnitine was obtained. The final product was titrated with carnitine acetyltransferase and found to contain 89.5% of L-carnitine hydrochloride. It was obtained with an aggregate yield of 57% with respect to dehydrocarnitine hydrochloride.