Polypeptide having diterpene synthase activity

An isolated polypeptide,

• the polypeptide comprising an amino acid sequence which comprises
• a sequence having at least 50% sequence identity to SEQ ID NO:
• 1,
• a sequence having at least 40% sequence identity to SEQ ID NO:
• 2, and
• at least one sequence selected from the group consisting of
• i) a sequence having at least 15% sequence identity to SEQ ID NO:
• 7;
• ii) a sequence having at least 25% sequence identity to SEQ ID NO:
• 4;
• iii) a sequence having at least 45% sequence identity to SEQ ID NO:
• 5;
• and
• iv) a sequence having at least 45% sequence identity to SEQ ID NO:
• 6,
• wherein SEQ ID NOs:
• 1-2 and 4-7 are of Clitopilus passeckerianus origin and wherein said polypeptide is a diterpene synthase.

An isolated nucleic acid molecule comprising a nucleotide sequence which

• has at least 40% sequence identity to SEQ ID NO:
• 15 or the sequence complementary thereto;
• or
• which has at least 60% sequence identity to SEQ ID NO:
• 8;
• or
• which has at least 60% sequence identity to a partial sequence of SEQ ID NO:
• 15 or the sequence complementary thereto, which partial sequence encodes a diterpene synthase.

An isolated nucleic acid molecule comprising

• (i) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence which comprises:
• a sequence having at least 50% sequence identity to SEQ ID NO:
• 1,
• a sequence having at least 40% sequence identity to SEQ ID NO:
• 2, and
• at least one sequence selected from the group consisting of
• a) a sequence having at least 15% sequence identity to SEQ ID NO:
• 7;
• b) a sequence having at least 25% sequence identity to SEQ ID NO:
• 4;
• c) a sequence having at least 45% sequence identity to SEQ ID NO:
• 5;
• and
• d) a sequence having at least 45% sequence identity to SEQ ID NO:
• 6, wherein SEQ ID NOs:
• 1-2 and 4-7 are of Clitopilus passeckerianus origin and wherein said polypeptide is a diterpene synthase
• or a polypeptide of SEQ ID NO:
• 9;
• or
• (ii) a nucleotide sequence which has at least 40% sequence identity to SEQ ID NO:
• 15 or the sequence complementary thereto;
• or
• (iii) a nucleotide sequence which has at least 60% sequence identity to SEQ ID NO:
• 8;
• or
• (iv) a nucleotide sequence which has at least 60% sequence identity to a partial sequence of SEQ ID NO:
• 15 or the sequence complementary thereto, which partial sequence encodes a diterpene synthase;
• or
• (v) a nucleotide sequence, which is capable of hybridizing to SEQ ID NO:
• 8, or SEQ ID NO:
• 13, or both under stringent conditions.

A method of producing pleuromutilin, the method comprising

• (i) introducing into a host selected from a cell, tissue and non-human organism a nucleic acid molecule having the sequence of SEQ ID NO:
• 15 or the sequence complementary thereto, or a vector comprising a nucleic acid molecule having the sequence of SEQ ID NO:
• 15 or the sequence complementary thereto, and
• (ii) cultivating the host under conditions suitable for the production of pleuromutilin.