Mikrobiol. Z. 2021; 83(5):19-29.
doi: https://doi.org/10.15407/microbiolj83.05.019

Production of Hydrogen by Purple Non-Sulfur Bacteria Rhodopseudomonas yavorovii IMV B-7620

O.V. Tarabas1, S.O. Hnatush1, O.B. Tashyrev2, V.M. Hovorukha2,
O.A. Havryliuk2, O.M. Moroz1, A.A. Halushka1

1Ivan Franko National University of Lviv
4 Hrushevsky Str., Lviv, 79005, Ukraine

2Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

Hydrogen production by microorganisms is studied by using different sources of carbon for their cultivation. Purple non-sulfur bacteria are capable of producing molecular hydrogen phototrophically with the simultaneous accumulation of biomass on organic substrates that may be waste from various industries. That fact makes the study of this group of microorganisms very much promising. The aim. The determination of the ability of purple non-sulfur bacteria Rhodopseudomonas yavorovii IMV B-7620 to produce hydrogen consuming different organic substrates and their effects on the main metabolic indicators of culture growth. Methods. Bacteria were grown in 100 mL glass jars in liquid modified ATCC No. 1449 medium for 14 days at temperature +27...+30ºC and at constant light (200 lux). Biomass accumulation and hydrogen production in the cultivation medium were determined using sodium acetate (12 and 36 mM), malate (12 and 36 mM), succinate (36 mM), glucose (36 mM), starch (36 mM), sodium citrate (36, 60, 90 mM). Biomass was determined turbidimetrically, the composition of the gas phase was determined using a gas chromatograph LHM-8-MD, redox potential and pH were estimated potentiometrically. The volume of gas synthesized was measured on a syringe scale. Determination of the content of organic acids in the culture liquid was analyzed by high performance liquid chromatography. Results. The utilization of organic compounds (malate, glucose, starch, sodium citrate) by R. yavorovii IMV B-7620 is accompanied by hydrogen synthesis. Under the growth with sodium acetate, bacteria produce small amounts of succinate. The malate metabolism results in the production of small amounts of fumarate on the 7th day of cultivation and isocitrate on the 10th day of cultivation. On the 14th day of cultivation, the cultural liquid contains a small amount of succinate. On the 14th day of cultivation, R. yavorovii IMV B-7620 produces 7.64±0.04% of hydrogen in the medium with malate (36 mM). However, the maximum concentration of hydrogen in the gas phase (21.26±0.08%) was gained on the 14th day of cultivation in the medium with sodium citrate. The maximum concentration of H2 in the gas phase during the growth in the medium with sodium citrate (60 mM) and NH4+ was 27.83±5.46% on the 7th day of cultivation and 35.69±0.40% with increasing concentration of sodium citrate up to 90 mM on the 10th day of cultivation. The total volume of hydrogen was 25.54±0.49 mL of H2 during the growth of R. yavorovii IMV B-7620 in the medium with 90 mM sodium citrate and NH4+. That is 1.5 times more than the amount of H2 produced during the growth of bacteria in the medium with 60 mM sodium citrate with the addition of NH4+. Conclusion. Purple non-sulfur bacteria R. yavorovii IMV B-7620 synthesize hydrogen during photofermentation of organic compounds. Bacteria were isolated from the water of Yavoriv Lake (Lviv region, Ukraine) formed as a result of flooding of the sulfur quarry. Bacteria consume sodium citrate, malate, glucose, starch and emit hydrogen. The total volume of hydrogen during the growth of R. yavorovii IMV B-7620 in the medium with 90 mM sodium citrate and NH4+ is 25.54±0.49 mL H2.

Keywords: hydrogen, purple non-sulfur bacteria, photofermentation.

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