Overexpression of acetyl-CoA synthetase increased the biomass and fatty acid proportion in microalga Schizochytrium.
High acetate accumulation was produced during glucose fermentation in high cell density cultures, which is harmful to cell growth. In order to reduce the negative impact of acetate accumulation on the fermentation products, we introduced the Escherichia coli acetyl-CoA synthetase (ACS) gene into the marine microalga Schizochytrium sp. TIO1101, generating genetically modified ACS transformants. The results of PCR and blotting analyses showed that the exogenous ACS gene was incorporated into the genome and successfully expressed. The engineered Schizochytrium increased the pH value and reduced the acetate concentration in the final fermentation medium significantly. Furthermore, the ACS transformants exhibited faster growth and glucose consumption rates than the wild-type strain. The biomass and fatty acid proportion of ACS transformants increased by 29.9 and 11.3 %, respectively. Taken together, the data suggest that ACS overexpression in Schizochytrium might improve the utilization of carbon resource and decrease the production of acetate byproduct. These results demonstrate that application of ACS in metabolic genetic engineering could improve the properties of Schizochytrium significantly.
School of Life Science and Biopharmaceutics, Shenyang Pharaceutical University, 103 Wenhua Road, Shenyang 110016, People's Republic of China.
Jinfei
J
Yan
Rubin
R
Cheng
Xiangzhi
X
Lin
Song
S
You
Ke
K
Li
Hui
H
Rong
Yong
Y
Ma
D000106:Acetate-CoA Ligase / Q000096:biosynthesis* / Q000235:genetics; D018533:Biomass; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005227:Fatty Acids / Q000378:metabolism*; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D058009:Stramenopiles / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D003470:Culture Media; D005227:Fatty Acids; D011994:Recombinant Proteins; D000106:Acetate-CoA Ligase; D005947:Glucose