Process for selection of Oxygen-tolerant algal mutants that produce H2 under aerobic conditions

摘要

A process for selection of oxygen-tolerant, H2-producing algal mutant cells comprising: (a) growing algal cells photoautotrophically under fluorescent light to mid log phase; (b) inducing algal cells grown photoautrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas; (2) incubating the suspension in the absence of light at ambient temperature; (c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light. (d) washing off metronidazole and sodium azide to obtain final cell suspension; (e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear; (f) counting the number of colonies to determine the percent of mutant survivors; and (g) testing survivors to identify oxygen-tolerant H2-producing mutants.