Synechocystis PCC 6803 overexpressing RuBisCO grow faster with increased photosynthesis

Metabolic Engineering Communications
Feiyan Liang, Peter Lindblad

Abstract

The ribulose-1,5-bisphosphate (RuBP) oxygenation reaction catalyzed by Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is competing with carboxylation, being negative for both energy and carbon balances in photoautotrophic organisms. This makes RuBisCO one of the bottlenecks for oxygenic photosynthesis and carbon fixation. In this study, RuBisCO was overexpressed in the unicellular cyanobacterium Synechocystis PCC 6803. Relative RuBisCO levels in the engineered strains FL50 and FL52 increased 2.1 times and 1.4 times, respectively, and both strains showed increased growth, photosynthesis and in vitro RuBisCO activity. The oxygen evolution rate increased by 54% and 42% on per chlorophyll basis, while the in vitro RuBisCO activity increased by 52% and 8.6%, respectively. The overexpressed RuBisCO were tagged with a FLAG tag, in strain FL50 on the N terminus of the large subunit while in strain FL52 on the C terminus of the small subunit. The presence of a FLAG tag enhanced transcription of the genes encoding RuBisCO, and, with high possibility, also enhanced the initiation of translation or stability of the enzyme. However, when using a streptavidin-binding tag II (strep-tag II), we did not observe a similar effect. Tagg...Continue Reading

Citations

Sep 7, 2017·FEMS Microbiology Letters·Angela ZhangShota Atsumi
Apr 28, 2020·The Journal of General and Applied Microbiology·Katsuhiko OkadaMikio Tsuzuki
Apr 14, 2017·FEMS Microbiology Letters·Han Min Woo, Hyun Jeong Lee
Jan 18, 2020·Plants·Valeria VecchiLuca Dall'Osto
Feb 6, 2021·Trends in Biotechnology·Mihris Ibnu Saleem NaduthodiMaria J Barbosa
Mar 18, 2021·Biotechnology Advances·Fei DuHe Huang

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Methods Mentioned

BETA
PCR
PCRs
protein assay

Software Mentioned

Quantity One

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