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Control of Cell Size and Nutrient Utilization by a Novel Regulator mraZ and Its Impact on Polyhydroxybutyrate (PHB)
Yuni Shin1, Taeyun Kim1, Ayeon Kim2
1Advanced Materials Program, Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea.
Abstract:
Cupriavidus necator is a well-established microbial platform for polyhydroxyalkanoate (PHA) production and numerous metabolic engineering strategies have been applied to improve its performance. However, little is known of the relationship between cell size and the production of the poly-3-hydroxybutyrate (PHB), with studies mainly focusing on cell division markers such as ftsZ, mreB, and minCD. These genes are influenced by multiple regulatory factors, though the specific regulators have not been characterized. This study investigated the role of a novel regulatory gene mraZ, which is located upstream of ftsZ and acts as a transcriptional repressor of cell growth and division, in PHA production by C. necator. We constructed an mraZ-deleted strain to assess the physiological and metabolic changes. qRT-PCR revealed the upregulated expression of ftsZ and PHA biosynthesis genes, along with ∼20% higher cell growth and PHA accumulation in the mutant across various culture conditions. Simultaneously, the cell length was reduced by >5 times that of the wild type, as confirmed by scanning and transmission electron microscopy. Although the smaller cell size increased the surface-to-volume ratio and enhanced the sugar utilization efficiency, it did not alter the molecular weight or mechanical properties of PHB. These findings highlight the value of targeting upstream regulators such as mraZ and provide new insights into the significance of reduced cell size in relation to PHB production. Considering that regulators constitute ∼12% of all genes in C. necator, our results highlight the untapped potential of regulator-based molecular engineering for optimizing PHA production.
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