Heterologous expression is an efficient approach to activate silent gene clusters of bioactive natural products.The rare actinomycetes strain Amycolatopsis sp.TNS106 is a producer of ristomycin A,which exhibits fast growth, short fermentation time and convenient genetic manipulation system.In order to develop this strain into a heterologous expression host for biosynthetic gene clusters (BGCs),the non-ribosomal peptide synthetase gene (rpsA) essential for the ristomycin A biosynthesis was replaced with a cassette containing the bacterial attachment sites attBΦC31 and attBΦBT1 via homologous recombination to construct a host with a clean background of secondary metabolism and two integrative sites.To test the obtained strain HXR1,integrative plasmids containing actinorhodin BGC from Streptomycescoelicolor or spinosad BGC from Saccharopolysporaspinosa were conjugated into HXR1.The results of fermentation and product analyses showed that actinorhodin and spinosad were successfully produced in HXR1.Compared with the Sac. erythraea?derived host LJ161 belonging to the non-Streptomyces actinomycete host,the actinorhodin production from HXR1 was approximately 1 day earlier and 1.3-fold higher.The Amycolatopsis sp.TNS106-derived host HXR1 will provide a useful platform for accelerating the discovery of novel secondary metabolites from Streptomyces and rare actinomycetes.
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胡欣瑞,贺卫军,吕金,王业民,陶美凤. Construction of an Amycolatopsis sp. TNS106 derived host and heterologous expression of secondary metabolic gene clusters[J]. Jorunal of Huazhong Agricultural University,2022,41(3):164-172.