期刊
BIOTECHNOLOGY LETTERS
卷 36, 期 4, 页码 821-827出版社
SPRINGER
DOI: 10.1007/s10529-013-1418-9
关键词
Algal lipids; Chlamydomonas reinhardtii; Escherichia coli; Gene expression; Lipid production; Phosphoenolpyruvate carboxylase; Reverse genetics
资金
- National High Technology Research and Development Program of China (863 Program) [2009AA064401]
- Shanghai Universities First-Class Disciplines Project [0707]
To investigate the function of a bacterial-type phosphoenolpyruvate carboxylase (PEPC2) derived from photosynthetically-grown Chlamydomonas reinhardtii, a fragment of the pepc2 gene was cloned and expressed in Escherichia coli. After optimal induction for 6 h, PEPC activity in the reverse mutant was lower than wild type (0.9 vs. 1.7 U/mg protein), and soluble protein was also lower than wild type (119 vs. 186 mg/g dry wt). In contrast, the total lipid content was increased from 56 (in wild type) to 71 mg/g dry wt, despite the growth rate being slightly diminished. The changes in PEPC activity, soluble protein and total lipid in the forward mutant were the opposite (2.4 U/mg, 230 mg/g, and 44 mg/g dry wt, respectively). Together, these data indicate that PEPC may function as a metabolic pivot in the regulation of protein and lipid accumulation in this alga.
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