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Author Lu, Wenyu ♦ Liu, Ting ♦ Zhang, Chuanbo
Source Directory of Open Access Journals (DOAJ)
Content type Text
Publisher BioMed Central
File Format HTM / HTML
Date Created 2018-08-20
Copyright Year ©2018
Language English
Subject Domain (in LCC) QR1-502
Subject Keyword Synthetic biology ♦ Levopimaric acid ♦ Levopimaradiene ♦ Science ♦ Microbiology ♦ Saccharomyces cerevisiae
Abstract Abstract Background 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. Results 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 400.31 mg/L via fed-batch fermentation in a 5-L bioreactor. Conclusions This is the first report to produce LA in a yeast cell factory and the highest titer of LA is achieved.
ISSN 14752859
Age Range 18 to 22 years ♦ above 22 year
Educational Use Research
Education Level UG and PG ♦ Career/Technical Study
Learning Resource Type Article
Publisher Date 2018-07-01
e-ISSN 14752859
Journal Microbial Cell Factories
Volume Number 17
Issue Number 1
Page Count 10
Starting Page 1
Ending Page 10

Source: Directory of Open Access Journals (DOAJ)