PT  - JOURNAL ARTICLE
AU  - Alexander Hammel
AU  - Frederik Sommer
AU  - David Zimmer
AU  - Mark Stitt
AU  - Timo Mühlhaus
AU  - Michael Schroda
TI  - Overexpression of sedoheptulose-1,7-bisphosphatase enhances photosynthesis in <em>Chlamydomonas reinhardtii</em> and has no effect on the abundance of other Calvin-Benson cycle enzymes
AID  - 10.1101/2020.02.14.949040
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.02.14.949040
4099  - http://biorxiv.org/content/early/2020/02/14/2020.02.14.949040.short
4100  - http://biorxiv.org/content/early/2020/02/14/2020.02.14.949040.full
AB  - The productivity of plants and microalgae needs to be increased to feed the growing world population and to promote the development of a low-carbon economy. This goal can be achieved by improving photosynthesis via genetic engineering. In this study, we have employed the Modular Cloning strategy to overexpress the Calvin-Benson cycle (CBC) enzyme sedoheptulose-1,7 bisphosphatase (SBP1) up to 3-fold in the unicellular green alga Chlamydomonas reinhardtii. The protein derived from the nuclear transgene represented ∼0.3% of total cell protein. Photosynthetic rate and growth were significantly increased in SBP1-overexpressing lines under high-light and high-CO2 conditions. Absolute quantification of the abundance of all other CBC enzymes by the QconCAT approach revealed no consistent differences between SBP1-overexpressing lines and the recipient strain. This analysis also revealed that the eleven CBC enzymes represent 11.9% of total cell protein in Chlamydomonas. Here the range of concentrations of CBC enzymes turned out to be much larger than estimated earlier, with a 128-fold difference between the most abundant CBC protein (rbcL) and the least abundant (triose phosphate isomerase). Accordingly, the concentrations of the CBC intermediates are often but not always higher than the binding site concentrations of the enzymes for which they act as substrates. The enzymes with highest substrate to binding site ratios might represent good candidates for overexpression in subsequent engineering steps.