Microalgae are small, single-celled, or simple multicellular organisms that contain Chlorophyll a, allowing them to efficiently convert CO₂ and water into organic matter through photosynthesis. They are valuable in producing a range of products such as biofuels, food, pharmaceuticals, and cosmetics, making them economically and environmentally significant. Currently, CO₂ is delivered to microalgae cultivation systems mainly through aeration with CO₂-enriched gases. However, this method demonstrates limited CO₂ absorption efficiency (13–20%), which reduces carbon utilization effectiveness and significantly increases carbon-source expenditure. To overcome these challenges, innovative CO₂ supplementation technologies have been introduced, raising CO₂ utilization rates to over 50%, accelerating microalgae growth, and reducing cultivation costs. This review first categorizes CO₂ supplementation technologies used in photobioreactor systems, focusing on different mechanisms for enhancing CO₂ mass transfer. It then evaluates the effectiveness of these technologies and explores their potential for scaling up. Among these strategies, membrane-based CO₂ delivery systems and the incorporation of CO₂ absorption enhancers have shown the highest efficiency in boosting CO₂ mass transfer and microalgae productivity. Future efforts should focus on integrating these methods into large-scale photobioreactor systems to optimize cost-effective, sustainable production.

Background: The collagenase encoding gene col was cloned into a pP43NMK vector and amplified in Escherichia coli JM109 cells. The shuttle vector pP43NMK was used to sub-clone the col gene to obtain the vector pP43NMK-col for the expression of collagenase in Bacillus subtilis WB600. The enzyme was characterized and the composition of the expression medium and culture conditions were optimized. Methods: The expressed recombinant enzyme was purified by ammonium sulfate, ultrafiltration, and through a nickel column. The purified collagenase had an activity of 9405.54 U/mg. Results: The recombinant enzyme exhibited optimal activity at pH 9.0 and 50 °C. Catalytic efficiency of the recombinant collagenase was inhibited by Fe³⁺ and Cu²⁺, but stimulated by Co²⁺, Ca²⁺, Zn²⁺, and Mg²⁺. The optimal conditions for its growth were at pH 7.0 and 35 °C, using 15 g/L of fructose and 36 g/L of yeast powder and peptone mixture (2:1) at 260 rpm with 11% inoculation. The maximal extracellular activity of the recombinant collagenase reached 2746.7 U/mL after optimization of culture conditions, which was 2.4-fold higher than that before optimization. Conclusions: This study is a first attempt to recombinantly express collagenase in B. subtilis WB600 and optimize its expression conditions, its production conditions, and possible scale-up.