Heterologous production of levopimaric acid in Saccharomyces cerevisiae

Microbial Cell Factories
Ting LiuWenyu Lu

Abstract

Levopimaric acid (LA), a type of diterpene resin acid produced by plants, is a significant industrial intermediate that is mainly produced via phytoextraction. This work aimed to apply synthetic biology to produce LA in yeast strains from a simple carbon source. Levopimaradiene (LP), the precursor of LA, was produced via LP synthase (LPS) expression in yeast. LPS was then modified by N-terminal truncating and site-directed mutagenesis. The strain containing t79LPSMM (79 N-terminal amino acid truncating and M593I/Y700F mutation) produced 6.92 mg/L of LP, which were 23-fold higher than the strain containing LPS. Next, t79LPSMM was expressed in a new metabolically engineered chassis, and the final LP production increased 164-folds to 49.21 mg/L. Three cytochrome P450 reductases (CPRs) were co-expressed with CYP720B1 (the enzyme responsible for LA production from LP) in yeast to evaluate their LA producing abilities, and the CPR from Taxus cuspidata (TcCPR) was found to be the best (achieved 23.13 mg/L of LA production). CYP720B1 and TcCPR genes overexpression in the multi-copy site of the S.cerevisiae genome led to a 1.9-fold increase in LA production to 45.24 mg/L in a shake-flask culture. Finally, LA production was improved to 4...Continue Reading

References

May 25, 2005·Proceedings of the National Academy of Sciences of the United States of America·Dae-Kyun RoJörg Bohlmann
Jan 17, 2008·Proceedings of the National Academy of Sciences of the United States of America·Christopher I KeelingJörg Bohlmann
Dec 17, 2008·Nucleic Acids Research·Zengyi ShaoHuimin Zhao
Jul 7, 2009·Annual Review of Plant Biology·James Kirby, Jay D Keasling
Nov 26, 2009·Phytochemistry·Kenneth Jensen, Birger Lindberg Møller
Apr 16, 2010·Applied Microbiology and Biotechnology·Chikara OhtoSakayu Shimizu
Jul 21, 2010·Proceedings of the National Academy of Sciences of the United States of America·Effendi LeonardKristala L J Prather
Oct 5, 2010·Natural Product Reports·Reuben J Peters
Sep 29, 2011·Nature Communications·Pamela P Peralta-YahyaTaek Soon Lee
Jan 6, 2012·The Journal of Biological Chemistry·Ke ZhouReuben J Peters
Jan 17, 2012·Proceedings of the National Academy of Sciences of the United States of America·Patrick J WestfallChris J Paddon
May 9, 2012·Biotechnology and Bioengineering·Zhubo DaiXueli Zhang
Aug 22, 2012·Natural Product Reports·Yang GaoReuben J Peters
Jun 6, 2013·Applied Biochemistry and Biotechnology·Ina NeunzigMatthias Bureik
Apr 9, 2014·Current Opinion in Plant Biology·Hugues RenaultDanièle Werck-Reichhart
Jul 6, 2014·The Plant Journal : for Cell and Molecular Biology·Philipp ZerbeJörg Bohlmann
Feb 1, 2015·Advances in Biochemical Engineering/biotechnology·Irini PaterakiBjörn Hamberger
Jun 18, 2016·Current Opinion in Chemical Biology·Ziga ZebecRainer Breitling
Apr 22, 2017·Critical Reviews in Biotechnology·Kalaivani Paramasivan, Sarma Mutturi
Jun 18, 2017·Current Opinion in Chemical Biology·Claudia E VickersJoel Cherry

❮ Previous
Next ❯

Citations

Sep 25, 2019·Microbial Cell Factories·Chuanbo ZhangWenyu Lu
Nov 19, 2020·Biotechnology and Bioengineering·Lihong JiangJiazhang Lian

❮ Previous
Next ❯

Datasets Mentioned

BETA
NP194183.1
CAA46815.1
AAT76449.1

Methods Mentioned

BETA
PCR

Software Mentioned

TASSER

Related Concepts

Related Feeds

Biomaterial Engineering

Advances in biomaterial engineering have permitted the development of sophisticated drug-releasing materials with a biomimetic 3D support that allow a better control of the microenvironment of transplanted cells. Here is the latest research.