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  • <div>In the present study, nanoscale organic-iron complex (NO-Fe) was used as an enhancement factor by two different <em>Rhodopseudomonas</em> species of purple non sulphur bacteria (PNSB) to produce hydrogen (H2). The NO-Fe complex was synthesised using <em>Eucalyptus viminalis</em>-a native Australian plant leaf extract, and FeSO4.7H2O salt. This NO-Fe complex was used as an iron source for newly isolated <em>Rhodopseudomonas palustris</em> MP3 and <em>Rhodopseudomonas harwoodiae</em> SP6 strains of photo-fermentative bacteria to produce H2.&nbsp;FeSO4.7H2O&nbsp;was also used as a source of iron for comparison with the NO-Fe complex. The photofermentative bacterial cultures were isolated from a fishpond, and only two strains, MP3 and SP6, were found viable after several attempts of quadrate streaking. After phylogenetic analysis, these strains were designated as <em>R. palustris</em> MP3 and <em>R. harwoodiae </em>SP6. The results showed that the <em>R. palustris</em> MP3 strain manifested approximately 70 % higher performance to the NO-Fe complex (FeEx1:2 and FeEx2:1), with an increase in H2 production compared to ferrous salt. The best performance was achieved by both strains when NO-Fe complex FeEx1:2 was supplemented in the fermentation broth at 10 mg/L concentration. The highest production of H2 was observed by <em>R. palustris</em> MP3 (10.7 ml/L) compared to <em>R. harwoodiae</em> SP6 (10 ml/L) when NO-Fe complex FeEx1:2 was used as an iron source. The study revealed that <em>R. palustris</em> MP3 and <em>R. harwoodiae</em> SP6 exhibited higher response to NO-Fe complex FeEx1:2 compared to the control. NO-Fe complex FeEx1:2 was considered highly conductive&nbsp;for efficient H2 production for further research.</div> <b>Citation:</b> Kanwal, F.; Tahir, A.; Tsuzuki, T.; Nisbet, D.; Chen, J.; Torriero, A.A.J. Comparison of Hydrogen Production Efficiency by <i>Rhodopseudomonas palustris MP3</i> and <i>Rhodopseudomonas harwoodiae SP6</i> Using an Iron Complex as an Enhancement Factor. <i>Energies</i> <b>2023</b>, <i>16</i>, 5018. https://doi.org/10.3390/en16135018