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Genome mining of 2-phenylethanol biosynthetic genes from Enterobacter sp. CGMCC 5087 and heterologous overproduction in Escherichia coli
- Source :
- Biotechnology for Biofuels, Vol 11, Iss 1, Pp 1-15 (2018)
- Publication Year :
- 2018
- Publisher :
- BMC, 2018.
-
Abstract
- Background 2-Phenylethanol (2-PE) is a higher aromatic alcohol that is widely used in the perfumery, cosmetics, and food industries and is also a potentially valuable next-generation biofuel. In our previous study, a new strain Enterobacter sp. CGMCC 5087 was isolated to produce 2-PE from glucose through the phenylpyruvate pathway. Results In this study, candidate genes for 2-PE biosynthesis were identified from Enterobacter sp. CGMCC 5087 by draft whole-genome sequence, metabolic engineering, and shake flask fermentation. Subsequently, the identified genes encoding the 2-keto acid decarboxylase (Kdc) and alcohol dehydrogenase (Adh) enzymes from Enterobacter sp. CGMCC 5087 were introduced into E. coli BL21(DE3) to construct a high-efficiency microbial cell factory for 2-PE production using the prokaryotic phenylpyruvate pathway. The enzymes Kdc4427 and Adh4428 from Enterobacter sp. CGMCC 5087 showed higher performances than did the corresponding enzymes ARO10 and ADH2 from Saccharomyces cerevisiae, respectively. The E. coli cell factory was further improved by overexpressing two upstream shikimate pathway genes, aroF/aroG/aroH and pheA, to enhance the metabolic flux of the phenylpyruvate pathway, which resulted in 2-PE production of 260 mg/L. The combined overexpression of tktA and ppsA increased the precursor supply of erythrose-4-phosphate and phosphoenolpyruvate, which resulted in 2-PE production of 320 mg/L, with a productivity of 13.3 mg/L/h. Conclusions The present study achieved the highest titer of de novo 2-PE production of in a recombinant E. coli system. This study describes a new, efficient 2-PE producer that lays foundation for the industrial-scale production of 2-PE and its derivatives in the future.
- Subjects :
- 0301 basic medicine
2-Keto acid decarboxylase
lcsh:Biotechnology
Enterobacter sp. CGMCC 5087
Management, Monitoring, Policy and Law
medicine.disease_cause
Applied Microbiology and Biotechnology
lcsh:Fuel
Metabolic engineering
03 medical and health sciences
chemistry.chemical_compound
Biosynthesis
lcsh:TP315-360
lcsh:TP248.13-248.65
medicine
Shikimate pathway
Overproduction
Escherichia coli
Alcohol dehydrogenase
chemistry.chemical_classification
biology
Renewable Energy, Sustainability and the Environment
Phenylpyruvate pathway
030104 developmental biology
General Energy
Enzyme
chemistry
Biochemistry
biology.protein
2-Phenylethanol
Fermentation
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 17546834
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biotechnology for Biofuels
- Accession number :
- edsair.doi.dedup.....872c7569a26e68be1ec37c75f691acf1