<P><B>Abstract</B></P> <P>Biohydrogen production from sugar beet molasses was investigated by using agar immobilized <I>R. capsulatus</I> YO3. A panel photobioreactor (1.4 L) was employed for a long-term hydrogen production in both indoor and outdoor conditions. The impact of several initial molasses concentrations on hydrogen production, yield and productivity were assessed. Indoor studies revealed that initial sucrose concentration in molasses should be kept below 20 mM to prevent inhibition of hydrogen production. The highest hydrogen productivity of 0.64 ± 0.06 mmol H<SUB>2</SUB> L<SUP>−1</SUP> h<SUP>−1</SUP> and yield of 12.2 ± 1.5 mol H<SUB>2</SUB>/mol sucrose were obtained in indoors throughout 20 days of operation. For outdoors, hydrogen production continued for 40 days including consecutive 10 rounds under natural outdoor conditions. In outdoor conditions, the maximum hydrogen productivity and yield were 0.79 ± 0.04 mmol H<SUB>2</SUB> L<SUP>−1</SUP> h<SUP>−1</SUP> and 5.2 ± 0.4 mol H<SUB>2</SUB>/mol sucrose respectively. These results indicate that the proposed system is promising for biohydrogen production from molasses at large-scale natural conditions.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Hydrogen production from molasses by agar immobilized <I>Rhodobacter capsulatus</I> YO3. </LI> <LI> Long-term hydrogen production was carried out in indoors and outdoors. </LI> <LI> High sucrose concentration in molasses inhibits H<SUB>2</SUB> production. </LI> <LI> Maximum hydrogen yield of 12.2 mol H<SUB>2</SUB>/mol sucrose was achieved in indoors. </LI> <LI> Maximum productivity of 0.79 mmol H<SUB>2</SUB> L<SUP>−1</SUP>h<SUP>−1</SUP> was achieved in outdoor conditions. </LI> </UL> </P>