Engineering Fungal Nonreducing Polyketide Synthase by Heterologous Expression and Domain Swapping

• Published in: Organic letters Vol. 15; no. 4; pp. 756 - 759
• Main Authors: Yeh, Hsu-Hua, Chang, Shu-Lin, Chiang, Yi-Ming, Bruno, Kenneth S, Oakley, Berl R, Wu, Tung-Kung, Wang, Clay C. C
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 15.02.2013; Amer Chemical Soc
• Subjects: aldehydes; Aldehydes - chemistry; Aspergillus nidulans - enzymology; Aspergillus, Polyketide Synthase, Domain Swapping, Thioesterase Domain; BASIC BIOLOGICAL SCIENCES; biosynthesis; carboxylic acids; Carboxylic Acids - chemistry; chemical reactions; chemical structure; Chemistry; Chemistry, Organic; chimerism; engineering; fungi; genes; heterologous gene expression; metabolites; Molecular Structure; Physical Sciences; polyketide synthases; Polyketide Synthases - genetics; Polyketide Synthases - metabolism; polyketides; Protein Engineering - methods; Science & Technology; SYSTEM; ASPERGILLUS-NIDULANS; BIOSYNTHESIS; GENES; RELEASE; IDENTIFICATION; CYCLIZATION
• ISSN: 1523-7060; 1523-7052; 1523-7052
• DOI: 10.1021/ol303328t

We reannotated the A. niger NR-PKS gene, e_gw1_19.204, and its downstream R domain gene, est_GWPlus_C_190476, as a single gene which we named dtbA. Heterologous expression of dtbA in A. nidulans demonstrated that DtbA protein produces two polyketides, 2,4-dihydroxy-3,5,6-trimethylbenzaldehyde (1) and 6-ethyl-2,4-dihydroxy-3,5-dimethylbenzaldehyde (2). Generation of DtbAΔR+TE chimeric PKSs by swapping the DtbA R domain with the AusA (austinol biosynthesis) or ANID_06448 TE domain enabled the production of two metabolites with carboxylic acids replacing the corresponding aldehydes.