Iterative analysis of metabolic modulation in the cyanobacterium Aphanizomenon flos-aquae 2012 KM1/D3 upon nitric oxide synthase derived NO induction

Abstract

Nitric oxide synthase (NOS) in mammals is recognized for its essentialities in several metabolism including blood vascular relaxation and nerves transmission etc. yet their functionalities in prokaryotes are largely unknown. Our study aimed to evaluate the putative role of nitric oxide synthase (NOS) derived nitric oxide (NO) during oxidative burst in the cyanobacterium, Aphanizomenon flos-aquae 2012/KM1/D3. Here, the accumulation of NO was dramatically reduced upon NOS inhibitor L-NG-Nitro arginine methyl ester (L-NAME) supplementation, exhibiting significant NO synthesis by NOS, whereas addition of L-arginine increase NOS derived NO in a dose dependent manner. Moreover, the reduction in the growth and metabolic activities of the cyanobacterium were evident upon L-NAME treatment that possibly pertained to decline in photopigments, PSII efficiency, loss in membrane integrity and DNA damage due to oxidative burst which culminated into cell death. Besides, the increment in carbohydrates and lipid content ensued with a decrease in protein content, indicating gluconeogenesis. Additionally, NOS inhibition disrupted the fatty acid and hydrocarbon profile, suggesting diminished membrane fluidity and cell integrity. However, higher content of flavonoids, phenolics, thiols and proline in L-NAME treated cells was also observed. Furthermore, L-arginine supplementation enhanced pigment content, photosynthetic efficiency, and reduced oxidative stress, thereby enhancing cyanobacterial growth. Further, L-arginine supplementation maintained Asc/DHAsc and GSH/GSSG ratio, conferred redox homeostasis. These results suggest that the NOS activity plays a critical role in protecting cyanobacteria from oxidative burst, maintaining their physiological balance.