Indolyl-3-alkane alpha-hydroxylase compositions from pseudomonas

Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;... – Enzyme or microbial cell is immobilized on or in an...

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435177, 435178, 435180, 435195, 4352533, 435259, 435802, 435814, 435818, 435874, C12N 1114, C12N 914, C12N 120, C12N 106

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057233250

ABSTRACT:
Compositions containing two species of indolyl-3-alkane alpha-hydroxylase (INDH) are isolated from Pseudomonas XA. An INDH1 composition contains protein subunits having molecular weights of 75,000, 34,500 and 32,500 daltons. An INDH2 composition contains protein subunits having molecular weights of 60,000, 44,000 and 42,000 daltons. Molecular weights are determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The compositions have a Specific INDH Activity of at least 10 international Units of INDH activity per milligram of protein, and contain less than 1 nanogram of endotoxin per International Unit of Specific INDH Activity. The INDH compositions may be immobilized on an insoluble matrix such as silica beads to provide at least 2.5 international Units of INDH activity per gram of Immobilized INDH composition. The INDH compositions are isolated by lysing Pseudomonas XA cells at a temperature of no more than 15.degree. C. to form a solution containing INDH, separating INDH from the solution at a temperature of no more than 15.degree. C. to form an isolated solution of INDH, determining protein concentration of the isolated solution, and if the protein concentration is less than 1 mg/ml, concentrating the solution such as by ultrafiltration to increase the protein concentration to within the range of 1 to 10 mg/ml.

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