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Title: Heterologous Gene Expression of N-Terminally Truncated Variants of LipPks1 Suggests a Functionally Critical Structural Motif in the N-terminus of Modular Polyketide Synthase

Journal Article · · ACS Chemical Biology
 [1];  [2];  [3];  [4];  [5];  [1];  [1];  [6];  [1];  [7];  [8]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Joint BioEnegy Institute, Emeryville, CA (United States); Agile BioFoundary, Emeryville, CA (United States)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. National Institute of Advanced Industrial Science and Technology, Higashi-hiroshima, Hiroshima (Japan)
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Agile BioFoundary, Emeryville, CA (United States)
  5. Agile BioFoundary, Emeryville, CA (United States)
  6. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Joint BioEnegy Institute, Emeryville, CA (United States)
  7. Univ. of California, Berkeley, CA (United States)
  8. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Joint BioEnegy Institute, Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Technical University of Denmark, Kogle, Allé (Denmark)
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Streptomyces genomes have a high G + C content and typically use an ATG or GTG codon to initiate protein synthesis. Although gene-finding tools perform well in low GC genomes, it is known that the accuracy in predicting a translational start site (TSS) is much less for high GC genomes. LipPks1 is a Streptomyces-derived, well-characterized modular polyketide synthase (PKS). Here, using this enzyme as a model, we experimentally investigated the effects of alternative TSSs using a heterologous host, Streptomyces venezuelae. One of the TSSs employed boosted the protein level by 59-fold and the product yield by 23-fold compared to the originally annotated start codon. Interestingly, a structural model of the PKS indicated the presence of a structural motif in the N-terminus, which may explain the observed different protein levels together with a proline and arginine-rich sequence that may inhibit translational initiation. Lastly, this structure was also found in six other modular PKSs that utilize noncarboxylated starter substrates, which may guide the selection of optimal TSSs in conjunction with start-codon prediction software.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1532274
Journal Information:
ACS Chemical Biology, Vol. 12, Issue 11; ISSN 1554-8929
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 6 works
Citation information provided by
Web of Science

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Cited By (2)

Technical Advances to Accelerate Modular Type I Polyketide Synthase Engineering towards a Retro-biosynthetic Platform journal June 2019
Short-chain ketone production by engineered polyketide synthases in Streptomyces albus journal November 2018

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