DNA contg. caffeine de-methylase gene - used in prodn. of 3-methyl-7-alkyl-xanthine cpds. by microbial demethylation of 1,3-di:methyl derivs.

摘要

A new isolated and purified DNA fragment (I) contains a caffeine demethylase (CDM) gene derived from a Pseudomonas sp. Also claimed are: (1) an isolated and purified DNA fragment contg. a base sequence coding for a defined sequence of 351 amino acids given in the specification; (2) recombinant DNA comprising a DNA fragment contg. a Pseudomonas CDM gene integrated into a vector; (3) a Pseudomonas transformant comprising a host transformed with recombinant DNA (2); (4) a process for producing a 3-methyl-7-alkyl-xanthino of formula (II) by culturing a transformant (3) in a nutrient medium contg. a 1,3-dimethyl-7-alkyl-xanthine of formula (III): R = alkyl; (5) a process for producing (11) by contacting (III) with a culture of a transformant (3), 'with microbial cells or a treatment product thereof', to form (II); (6) a process for producing 3-methyl-7-propyl-xanthine (IIa) by culturing a Pseudomonas sp. or mutant capable of converting 1,3-dimethyl-7-propyl-xanthine (IIIa) to (IIa), in a nutrient medium contg. (IIIa); (7) a process for producing (IIa) by contacting (IIIa) with a culture of a Pseudomonas sp. or mutant capable of converting (IIIa) to (IIa), 'with microbial cells or a treatment product thereof', to form (IIa). USE - (II) are useful as pharmaceutical intermediates: theobromine is an intermediate for 1-(5-oxohelyl)-3,7-dimethyl-xanthine (pentoxiphylline) and (IIa) is an intermediate for 1-(5-oxohexyl)-3-methyl-7-propyl-xanthine(propentophylline), which is useful for treating cerebrovascular disorders.