Metabolic engineering of Escherichia coli for high-level astaxanthin production with high productivity

Cited 36 time in webofscience Cited 0 time in scopus
  • Hit : 99
  • Download : 0
Astaxanthin is a reddish keto-carotenoid classified as a xanthophyll found in various microbes and marine organisms. As a powerful antioxidant having up to 100 times more potency than other carotenoids such as beta-carotene, lutein, and lycopene, astaxanthin is a versatile compound utilized in animal feed, food pigment, health promotion and cosmetic industry. Here, we report development of metabolically engineered Escherichia coli capable of producing astaxanthin to a high concentration with high productivity. First, the heterologous crt genes (crtE, crtY, crtI, crtB, and crtZ) from Pantoea ananatis and the truncated BKT gene (trCrBKT) from Chlamydomonas reinhardtii were introduced to construct the astaxanthin biosynthetic pathway. Then, eight different fusion tags were examined by attaching them to the N-or C-terminus of the trCrBKT membrane protein to allow stable expression and to efficiently guide trCrBKT to the E. coli membrane. When the signal peptide of OmpF and TrxA were tagged to the N-terminus and C-terminus of trCrBKT, respectively, astaxanthin production reached 12.90 mg/L (equivalent to 3.84 mg/gDCW), which was 2.08-fold higher than that obtained without tagging. Upon optimization of culture conditions, this engineered strain WLGB-RPP harboring pAX15 produced 332.23 mg/L (5.38 mg/gDCW) of astaxanthin with the productivity of 3.79 mg/L/h by fed-batch fermentation. In order to further increase astaxanthin production, in silico flux variability scanning based on enforced objective flux (FVSEOF) was performed to identify gene overexpression targets. The engineered strain WLGB-RPP (pAX15, pTrc-ispDF) which simultaneously overexpressing the ispD and ispF genes identified by FVSEOF produced astaxanthin to a higher concentration of 377.10 mg/L (6.26 mg/gDCW) with a productivity of 9.20 mg/L/h upon induction with 1mM IPTG. When cells were induced with 0.5 mM IPTG to reduce the metabolic burden, astaxanthin concentration further increased to 432.82 mg/L (7.12 mg/gDCW) with a productivity of 9.62 mg/L/h. To more stably maintain plasmid during the fed-batch fermentation of WLGB-RPP (pAX15, pTrc-ispDF), the posts-egregational killing hok/sok system was introduced. This strain produced 385.04 mg/L (6.98 mg/gDCW) of astaxanthin with a productivity of 7.86 mg/L/h upon induction with 0.5mM IPTG. The strategies reported here will be useful for the enhanced production of astaxanthin and related carotenoid products by engineered E. coli strains.
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Issue Date
2018-09
Language
English
Article Type
Article
Citation

METABOLIC ENGINEERING, v.49, pp.105 - 115

ISSN
1096-7176
DOI
10.1016/j.ymben.2018.08.002
URI
http://hdl.handle.net/10203/246581
Appears in Collection
CBE-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡ Click to see webofscience_button
⊙ Cited 36 items in WoS Click to see citing articles in records_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0