Rapid Discovery and Engineering of Natural Product Biosynthetic Pathways Via Synthetic Biology

The majority of existing antibacterial and anticancer drugs are natural products or their derivatives.  However, the discovery and sustainable production of these compounds are often hampered owing to limited ability to manipulate the corresponding biosynthetic pathways.  In this talk, I will discuss our recent efforts in developing synthetic biology approaches to address these overwhelming challenges in the pharmaceutical industry.  Specifically, I will present a genomics-driven, synthetic biology-based method, PANDA (Pathway chAracterization and eNgineering via DNA Assembler), for discovery, characterization, and engineering of new natural product biosynthetic pathways.  By taking advantage of the highly efficient yeast in vivo homologous recombination mechanism, this method synthesizes the entire expression vector containing the target biosynthetic pathway and the genetic elements needed for DNA maintenance and replication in Saccharomyces cerevisiae, Escherichia coli, and a target heterologous expression host in a single-step manner.  By using three distinct biosynthetic pathways, we demonstrate that PANDA offers unprecedented flexibility and versatility in pathway manipulations such as heterologous expression, introduction of single or multiple point mutations, scar-less gene deletion and replacement, and generation of new products.  This method should be generally applicable to any biochemical pathways.  In addition, I will show how a similar approach can be used to decipher cryptic biosynthetic pathways from sequenced microbial genomes and metagenomes.  Such pathways are widely considered as hidden treasures for discovery of novel antibacterial and anticancer drugs. 

Related references:

1. Z. Shao, H. Zhao, and H. Zhao. “DNA Assembler, an in vivo Genetic Method for Rapid Construction of Biochemical Pathways.” Nucleic Acids Research, 37, e16 (2009).

2. J. Liang, Y. Luo, and H. Zhao. “Synthetic Biology: Put Synthesis in Biology.” Wiley Interdisciplinary Reviews: Systems Biology and Medicine, in press.

3. Z. Shao, Y. Luo, and H. Zhao. “Rapid Characterization and Engineering of Natural Product Biosynthetic Pathways via DNA Assembler.” submitted.