Screening of broad-host expression promoters for shuttle expression vectors in non-conventional yeasts and bacteria.
Microb Cell Fact
; 23(1): 230, 2024 Aug 16.
Article
in En
| MEDLINE
| ID: mdl-39152436
ABSTRACT
BACKGROUND:
Non-conventional yeasts and bacteria gain significance in synthetic biology for their unique metabolic capabilities in converting low-cost renewable feedstocks into valuable products. Improving metabolic pathways and increasing bioproduct yields remain dependent on the strategically use of various promoters in these microbes. The development of broad-spectrum promoter libraries with varying strengths for different hosts is attractive for biosynthetic engineers.RESULTS:
In this study, five Yarrowia lipolytica constitutive promoters (yl.hp4d, yl.FBA1in, yl.TEF1, yl.TDH1, yl.EXP1) and five Kluyveromyces marxianus constitutive promoters (km.PDC1, km.FBA1, km.TEF1, km.TDH3, km.ENO1) were selected to construct promoter-reporter vectors, utilizing α-amylase and red fluorescent protein (RFP) as reporter genes. The promoters' strengths were systematically characterized across Y. lipolytica, K. marxianus, Pichia pastoris, Escherichia coli, and Corynebacterium glutamicum. We discovered that five K. marxianus promoters can all express genes in Y. lipolytica and that five Y. lipolytica promoters can all express genes in K. marxianus with variable expression strengths. Significantly, the yl.TEF1 and km.TEF1 yeast promoters exhibited their adaptability in P. pastoris, E. coli, and C. glutamicum. In yeast P. pastoris, the yl.TEF1 promoter exhibited substantial expression of both amylase and RFP. In bacteria E. coli and C. glutamicum, the eukaryotic km.TEF1 promoter demonstrated robust expression of RFP. Significantly, in E. coli, The RFP expression strength of the km.TEF1 promoter reached â¼20% of the T7 promoter.CONCLUSION:
Non-conventional yeast promoters with diverse and cross-domain applicability have great potential for developing innovative and dynamic regulated systems that can effectively manage carbon flux and enhance target bioproduct synthesis across diverse microbial hosts.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Kluyveromyces
/
Promoter Regions, Genetic
/
Yarrowia
/
Escherichia coli
/
Genetic Vectors
Language:
En
Journal:
Microb Cell Fact
Journal subject:
BIOTECNOLOGIA
/
MICROBIOLOGIA
Year:
2024
Document type:
Article
Affiliation country: