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Biochemistry & Molecular Biology Journal
ISSN: 2471-8084
Internat i ona l Conference on
Biotechnology, Biomarkers
& Systems Biology
M a r c h 0 4 - 0 5 , 2 0 1 9
Am s t e r d a m , N e t h e r l a n d s
Biotechnology, Biomarkers & Systems Biology 2019
T
erpenoids represent the largest class of natural productswith a diverse array
of structures and functions. Many terpenoids have reported therapeutic
properties such as antimicrobial, anti-inflammatory, immunomodulatory and
chemotherapeutic properties making them of great interest in the medical
field. Terpenoids suffer from low natural yields and complicated chemical
synthesis; hence there is a need for a more sustainable production method.
Metabolic engineering using biosynthetic mevalonate and non-mevalonate
pathways provides an excellent opportunity to construct microbial cell
factories producing terpenoids. The complexity and diversity of terpenoid
structures depends mainly on the action of the terpene synthases responsible
for their synthesis. Amorpha- 4, 11-diene synthase (ADS) cyclizes the substrate
farnesyl pyrophosphate to produce amorpha- 4, 11-diene as major product.
This is considered the first committed and rate-limiting step in the biosynthesis
of the antimalarial artemisinin. Here, we utilize a reported 3D model of ADS to
perform mutability landscape guided enzyme engineering. A mutant library of
258 variants along sixteen active site residues was created and then screened
for catalytic activity and product profile. This allowed for identification of the
role of some of these residues in the mechanism. The mutability landscape
also helped to identify variants with improved catalytic activity. H448A showed
~4 fold increase in catalytic efficiency and the double mutation T399S/H448A
showed that kcat has improved by ~5 times. This variant can be used to
enhance amorphadiene production and in turn artemisinin biosynthesis. Our
findings provide the basis for the first step in improving industrial production
of artemisinin and they open up possibilities for further engineering and
understanding of ADS.
Biography
Wim J Quax was appointed as Professor of Pharmaceutical
Biology at the University of Groningen in 1998. He has
developed an extensive research group focussing on directed
evolution and protein design technology for researching
pharmaceutically relevant proteins. One of his focus areas are
enzymes catalysing the synthesis of natural products. He has
published >300 peer reviewed papers and book chapters and he
is Inventor of >30 patents. He is the former Scientific Director of
the Groningen Research Institute for Pharmacy (GRIP).
w.j.quax@rug.nlEngineering the biosynthesis of artemisinin
Wim J Quax, Ingy I Abdallah, Hegar Pramastya and Dan Xue
University of Groningen, The Netherlands
Wim J Quax et al., Biochem Mol biol J 2019, Volume:5
DOI: 10.21767/2471-8084-C1-023