Abstract
This study investigates the enhancement of bioethanol production using a genetic engineering approach. The bioethanol-producing strain, E. aerogenes ATCC 29007, was engineered by deleting the D-lactate dehydrogenase (ldhA) gene to block the production of lactic acid. The Open-reading frame coding region of ldhA gene was replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method to inactivate chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. The colony PCR was used to confirm the deleted gene. Glycerol, a useful byproduct in the biodiesel industry, was employed to convert into bioethanol, using engineered E. aerogenes SUMI014. Under optimal conditions of fermentation (34 °C, pH 7.5, 78 h), bioethanol production by the mutant strain was 34.54 g/L, 1.5 times greater than that produced by its wild type (13.09 g/L). Subsequent overexpression of alcohol dehydrogenase (adhE) gene in the mutant strain; increased the production of bioethanol up to 38.32 g/L. By the combination of gene deletion and overexpression, the bioethanol yield was 0.48 g/g when employing 80 g/L glycerol. Hence, a significant enhancement in ethanol production was observed. These results may provide valuable guidelines for further engineering bioethanol producers.
| Original language | English |
|---|---|
| Pages (from-to) | 2051-2060 |
| Number of pages | 10 |
| Journal | Process Biochemistry |
| Volume | 50 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 2015 Dec 1 |
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Keywords
- Alcohol dehydrogenase
- d-lactate dehydrogenase
- Enterobacter aerogenes ATCC29007
- Enterobacter aerogenes SUMI014
- Enterobacter aerogenes SUMI2008
ASJC Scopus subject areas
- Biochemistry
- Applied Microbiology and Biotechnology
- Bioengineering
Cite this
Improved bioethanol production from metabolic engineering of Enterobacter aerogenes ATCC 29007. / Thapa, Laxmi Prasad; Lee, Sang Jun; Yang, Xiaoguang; Lee, Ju Hun; Choi, Han Suk; Park, Chulhwan; Kim, Seung Wook.
In: Process Biochemistry, Vol. 50, No. 12, 01.12.2015, p. 2051-2060.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Improved bioethanol production from metabolic engineering of Enterobacter aerogenes ATCC 29007
AU - Thapa, Laxmi Prasad
AU - Lee, Sang Jun
AU - Yang, Xiaoguang
AU - Lee, Ju Hun
AU - Choi, Han Suk
AU - Park, Chulhwan
AU - Kim, Seung Wook
PY - 2015/12/1
Y1 - 2015/12/1
N2 - This study investigates the enhancement of bioethanol production using a genetic engineering approach. The bioethanol-producing strain, E. aerogenes ATCC 29007, was engineered by deleting the D-lactate dehydrogenase (ldhA) gene to block the production of lactic acid. The Open-reading frame coding region of ldhA gene was replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method to inactivate chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. The colony PCR was used to confirm the deleted gene. Glycerol, a useful byproduct in the biodiesel industry, was employed to convert into bioethanol, using engineered E. aerogenes SUMI014. Under optimal conditions of fermentation (34 °C, pH 7.5, 78 h), bioethanol production by the mutant strain was 34.54 g/L, 1.5 times greater than that produced by its wild type (13.09 g/L). Subsequent overexpression of alcohol dehydrogenase (adhE) gene in the mutant strain; increased the production of bioethanol up to 38.32 g/L. By the combination of gene deletion and overexpression, the bioethanol yield was 0.48 g/g when employing 80 g/L glycerol. Hence, a significant enhancement in ethanol production was observed. These results may provide valuable guidelines for further engineering bioethanol producers.
AB - This study investigates the enhancement of bioethanol production using a genetic engineering approach. The bioethanol-producing strain, E. aerogenes ATCC 29007, was engineered by deleting the D-lactate dehydrogenase (ldhA) gene to block the production of lactic acid. The Open-reading frame coding region of ldhA gene was replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method to inactivate chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. The colony PCR was used to confirm the deleted gene. Glycerol, a useful byproduct in the biodiesel industry, was employed to convert into bioethanol, using engineered E. aerogenes SUMI014. Under optimal conditions of fermentation (34 °C, pH 7.5, 78 h), bioethanol production by the mutant strain was 34.54 g/L, 1.5 times greater than that produced by its wild type (13.09 g/L). Subsequent overexpression of alcohol dehydrogenase (adhE) gene in the mutant strain; increased the production of bioethanol up to 38.32 g/L. By the combination of gene deletion and overexpression, the bioethanol yield was 0.48 g/g when employing 80 g/L glycerol. Hence, a significant enhancement in ethanol production was observed. These results may provide valuable guidelines for further engineering bioethanol producers.
KW - Alcohol dehydrogenase
KW - d-lactate dehydrogenase
KW - Enterobacter aerogenes ATCC29007
KW - Enterobacter aerogenes SUMI014
KW - Enterobacter aerogenes SUMI2008
UR - http://www.scopus.com/inward/record.url?scp=84949110411&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84949110411&partnerID=8YFLogxK
U2 - 10.1016/j.procbio.2015.09.007
DO - 10.1016/j.procbio.2015.09.007
M3 - Article
AN - SCOPUS:84949110411
VL - 50
SP - 2051
EP - 2060
JO - Process Biochemistry
JF - Process Biochemistry
SN - 1359-5113
IS - 12
ER -