Heterologous expression of LamA gene encoded endo-β-1,3-glucanase and CO₂ fixation by bioengineered Synechococcus sp. PCC 7002

Abstract

Maximum growth rate of Synechococcus mutant was 0.083 h⁻¹ with 5% CO₂.

Maximum biomass concentration of Synechococcus mutant was 3.697 g·L⁻¹.

Synechococcus mutant can tolerate gas aeration with 15% CO₂.

Maximum specific activity of laminarinase was 4.325 U·mg⁻¹ dry mass.

Optimal pH and temperature of laminarinase activity were 8.0 and 70°C.

The gene for the catalytic domain of thermostable endo-β-1,3-glucanase (laminarinase) LamA was cloned from Thermotoga maritima MSB8 and heterologously expressed in a bioengineered Synechococcus sp. PCC 7002. The mutant strain was cultured in a photobioreactor to assess biomass yield, recombinant laminarinase activity, and CO2 uptake. The maximum enzyme activity was observed at a pH of 8.0 and a temperature of 70°C. At a CO2 concentration of 5%, we obtained a maximum specific growth rate of 0.083 h⁻¹, a biomass productivity of 0.42 g·L⁻¹·d⁻¹, a biomass concentration of 3.697 g·L⁻¹, and a specific enzyme activity of the mutant strain of 4.325 U·mg⁻¹ dry mass. All parameters decreased as CO2 concentration increased from 5% to 10% and further to 15% CO2, except enzyme activity, which increased from 5% to 10% CO2. However, the mutant culture still grew at 15% CO2 concentration, as reflected by the biomass productivity (0.26 g·L⁻¹·d⁻¹), biomass concentration (2.416 g·L⁻¹), and specific enzyme activity (3.247 U·mg⁻¹ dry mass).

Graphical abstract

Keywords

Synechococcus sp. PCC 7002 / Thermotoga maritima / LamA gene / Endo-β-1 / 3-glucanase / CO₂ fixation

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Acknowledgements

The authors are grateful to the Natural Science and Engineering Research Council (NSERC) of Canada for the financial support via a strategic partnership grant (#380768-09). Di Li also thanks the China Scholarship Council for a CSC PhD scholarship(#200001).

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