Avermectin aglycon synthase genes
An avermectin and aglycone technology, which can be used in genetic engineering, plant genetic improvement, carbohydrate active ingredients, etc., and can solve the problems of unknown nucleotide sequence and difficulty in breeding.
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example 1
[0400] example 1: Determination of the Nucleotide Sequence of the Avermectin Aglycone Synthase Gene of Streptomyces avermitilis
[0401] The nucleotide sequence of a DNA encoding avermectin aglycone synthase derived from Streptomyces avermitilis was determined as follows (US5206155, FERM BP-2773).
[0402] from genes containing the avermectin aglycone synthase genes (AveAI and AveAII) that co-segregated with the gene encoding the avermectin B5-O-methyltransferase (AveD; Genes, 206, 175-180, 1998). Contiguous or overlapping DNA fragments of the avermectin aglycone synthase gene were subcloned in fragmented plasmids. The nucleotide sequence of the inserted DNA fragment in the subclone is then determined.
[0403] More specifically, the complete nucleotide sequences of AveAI and AveAII can be determined by the following method: Subclones exist in figure 1 3.4kbp, 2.0kbp, 0.5kbp, 6.8kbp, 7.0kbp, 7.8kbp, 3.7kbp, 4.8kbp, 1.3kbp, 2.4kbp, 0.7kbp, 1.0kbp, 5.4kbp, BamHI-treated fra...
example 2
[0404] Example 2: Production of avermectin alone by nucleotide modification of the dehydratase domain on module 2 B2a
[0405] Streptomyces avermitilis K2033 (FERM BP-2775) can produce avermectin B1a and avermectin B2a.
[0406] The information on the avermectin aglycone synthase gene (SEQ ID NO: 1 and 2) derived from the avermectin-producing strain obtained and sequenced in Example 1 shows that avermectin is produced by figure 2 Biosynthetic pathways indicated.
[0407] Avermectin B 1a differs from avermectin B2a only in that avermectin B1a has a double bond between the 22- and 33-positions on its aglycone moiety, whereas avermectin B2a There is a single bond between the 22- and 33-positions on the aglycone moiety, and there is a hydroxyl group at the 23-position.
[0408] The avermectin biosynthetic pathways discovered above indicate that the avermectin aglycone synthase domain DH of SU2(DH2) determines between the 22- and 23-positions of the avermectin moiety of aver...
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