摘要:
The invention relates to modified cytochrome P450 monooxygenases comprising a modified substrate profile, to nucleic acid sequences that code therefor, to expression constructs and vectors, and to recombinant microorganisms which contain these vectors. The invention also relates to methods for microbiologically producing terminally or subterminally hydroxylated aliphatic carboxylic acids.
摘要:
A process for the mutagenesis of a double-stranded polynucleotide sequence (master sequence) of n base-pairs having a (+)-strand and a complementary (-)-strand comprising the steps (i) creation of a collection of single-stranded fragments of the (+)-strand of the master sequence wherein all members of the collection have the same 5'-terminus and have a deletion in the 3'-terminus such that the collection represents (+)-strands with a length of n-1, n-2, n-3, ..... nucleotides; (ii) introduction of at least one universal or degenerate nucleotide at the 3'-terminus of the (+) strand produced in step (i); (iii) elongation of the (+)-strand produced in step (ii) to the full length of the master sequence using the (-)-strand or fragments thereof as a template strand for the elongation; (iv) synthesis of a (-)-strand by using the (+)-strand produced in step (iii) as a template strand thereby effecting mutations in the (-)-strand at the positions of the previous universal or degenerate nucleotides compared to the master sequence.
摘要:
The invention relates to modified cytochrome P450 monooxygenases comprising a modified substrate profile, to nucleic acid sequences that code therefor, to expression constructs and vectors, and to recombinant microorganisms which contain these vectors. The invention also relates to methods for microbiologically producing terminally or subterminally hydroxylated aliphatic carboxylic acids.
摘要:
A process for the mutagenesis of a double-stranded polynucleotide sequence (master sequence) of n base-pairs having a (+)-strand and a complementary (-)-strand comprising the steps (i) creation of a collection of single-stranded fragments of the (+)-strand of the master sequence wherein all members of the collection have the same 5'-terminus and have a deletion in the 3'-terminus such that the collection represents (+)-strands with a length of n-1, n-2, n-3, ..... nucleotides; (ii) introduction of at least one universal or degenerate nucleotide at the 3'-terminus of the (+) strand produced in step (i); (iii) elongation of the (+)-strand produced in step (ii) to the full length of the master sequence using the (-)-strand or fragments thereof as a template strand for the elongation; (iv) synthesis of a (-)-strand by using the (+)-strand produced in step (iii) as a template strand thereby effecting mutations in the (-)-strand at the positions of the previous universal or degenerate nucleotides compared to the master sequence.
摘要:
The invention relates to a novel electron donor system for enzymes with redox properties and to its use in enzyme-catalyzed oxidation reactions, such as the production of φ hydroxylated fatty acids. The invention further relates to an improved analysis method for fatty acid monoxygenases and to bioreactors and test kits wherein the inventive electron donor system is advantageously used.
摘要:
The invention relates to novel cytochrome P450 monooxygenases comprising a modified substrate specificity, to nucleotide sequences which code therefor, to expression constructs and vectors containing these sequences, and to microorganisms transformed therewith. The invention also relates to methods for microbiologically oxidizing different organic substrates, such as methods for producing indigo and indirubin.
摘要:
The invention relates to a novel electron donor system for enzymes with redox properties and to its use in enzyme-catalyzed oxidation reactions, such as the production of φ hydroxylated fatty acids. The invention further relates to an improved analysis method for fatty acid monoxygenases and to bioreactors and test kits wherein the inventive electron donor system is advantageously used.