Accellular synthesis of cephalosporins

Claim 2

A process for producing a cephalosporin antibiotic, said process comprising the steps of: providing a cell-free extract of Cephalosporium acremonium; providing a source of starting material comprising a substance having the formula: ##STR7## wherein R is a hydrocarbon containing 1-4 carbon atoms terminally substituted with a radical selected from the group consisting of carboxy, amino, and combinations thereof; contacting the extract and the starting material in a reaction zone while promoting oxygen transfer; providing ATP as an energy source to said reaction zone; and allowing a component of the extract to react with said precursor for a sufficient amount of time to produce a cephalosporin antibiotic.

Claim 3

The process as set forth in claim 1 wherein the ATP utilized in the cephalosporin antibiotic production is regenerated by an ATP regenerating system comprising a phosphate donor and a phosphotransferease enzyme.

Claim 4

The process as set forth in claim 2 wherein the phosphate donor is phosphoenolpyruvate and the phosphotransferase enzyme is pyruvate kinase.

Claim 5

The process as set forth in claim 1 wherein said cell-free extract is made by treating Cephalosporium acremonium cells with a lysing enzyme.

Claim 6

The process as set forth in claim 4 wherein said cell-free extract is made by treating Cephalosporium acremonium cells with endoβ(1→3) glucanase and endoβ(1→4) glucanase and zymolyase.

Claim 7

The process as set forth in claim 1 wherein high oxygen transfer is promoted by vigorously shaking the reaction components in the reaction zone.

Claim 8

The process as set forth in claim 1 wherein mannitol and trace concentrations of KCL and MgSO 4 are included in the reaction and the system is buffered to about pH 7.2.

Claim 9

The process as set forth in claim 1 wherein the starting material is selected from the group consisting of substances having the formula: ##STR8## wherein R is selected from the group consisting of: --(CH.sub.2).sub.3 --COOH ##STR9##

Claim 10

The process as set forth in claim 8 wherein R is ##STR10##

Claim 11

A process for producing a cephalosporin antibiotic, said process comprising the steps of: providing a source of Cephalosporium acremonium mycelia; treating said mycelia with lytic enzyme endoβ(1→3) glucanse, endoβ(1→4) glucanse and, from Arthrobacter, zymolyase; incubating the treated mycelia to produce a protoplast suspension and homogenizing the suspension to produce a cell free extract; providing a source of starting material selected from the group consisting of: ##STR11## wherein R is: --(CH.sub.2).sub.3 --COOH ##STR12## contacting the extract and the starting material in the presence of ATP in a reaction zone while promoting oxygen transfer; and allowing a component of the extract to react with the starting material for a sufficient amount of time to produce a cephalosporin antibiotic.

Claim 12

A process for producing a cephalosporin antibiotic, said process comprising the steps of: providing a source of Cephalosporium acremonium mycelia; treating said mycelia with lytic enzyme endoβ(1→3) glucanse, endoβ(1→4) glucanse and, from Arthrobacter, zymolyase; incubating the treated mycelia to produce a protoplast suspension and homogenizing the suspension to produce a cell free extract; providing a source of starting material selected from the group consisting of: ##STR11## wherein R is: --(CH.sub.2).sub.3 --COOH ##STR12## contacting the extract and the starting material in the presence of ATP in a reaction zone while promoting oxygen transfer; and allowing a component of the extract to react with the starting material for a sufficient amount of time to produce a cephalosporin antibiotic.