Influence of fermentation reactions on continuous hydrogen photoproduction by microalga Chlamydomonas reinhardtii under sulfur deficiency
Abstract
The study investigated the effect of the major fermentation enzyme – pyruvate formate lyase – on hydrogen photoproduction by green microalga Chlamydomonas reinhardtii under mineral deprivation. Cultures of the wild type (CC-125) and mutant pfl1 without pyruvate formate lyase activity were incubated on a medium without sulfur in the sealed photoreactors under constant illumination for 96 h. During experiment, accumulation of hydrogen in gas phase of photoreactors and factors affecting hydrogen production, such as photosynthetic activity, respiration, and starch content in cells were assessed. Both strains showed similar photosynthetic activity and starch content when cultivated in the complete medium. Under sulfur depletion, the cultures of the pfl1 mutant began to evolve hydrogen later than the wild type; however, this process lasted much longer in the mutant. The pfl1 cultures showed higher rates of starch accumulation and breakdown at the initial and late stages of sulfur deprivation, respectively. Moreover, the mutant was characterized by higher photosynthetic activity at the later stage of starvation. The obtained results indicate close relationships between photosynthetic electron transport, hydrogenase reaction, carbohydrate metabolism and fermentation process, pointing to the prospect of using genetic engineering to modify fermentation reactions in order to improve photosynthetic hydrogen production in biotechnology.
Keywords
About the Authors
A. A. VolgushevaRussian Federation
Faculty of Biology
1–12 Leninskie Gory, Moscow, 119234
E. V. Petrova
Russian Federation
Faculty of Biology
1–12 Leninskie Gory, Moscow, 119234
G. P. Kukarskikh
Russian Federation
Faculty of Biology
1–12 Leninskie Gory, Moscow, 119234
A. Dubini
Spain
Faculty of Natural Sciences
Edif. Severo Ochoa, Campus de Rabanales, 14071, Córdoba
T. K. Antal
Russian Federation
21 Sovetskaya Street, Pskov, 180000
References
1. Antal T.K. The metabolic acclimation of Chlamydomonas reinhardtii to depletion of essential nutrients: application for hydrogen production // Microalgal Hydrogen Production: Achievements and Perspectives / Eds. M. Seibert, G. Torzillo. RSC Publishing, 2018. P. 235–264.
2. Kosourov S., Böhm M., Senger M., Berggren G., Stensjö K., Mamedov F., Lindblad P., Allahverdiyeva Y. Photosynthetic hydrogen production: novel protocols, promising engineering approaches and application of semi-synthetic hydrogenases // Physiol. Plant. 2021. Vol. 173. N 2. P. 555–567.
3. Antal T., Petrova E., Slepnyova V., Kukarskikh G., Volgusheva A., Dubini A., Baizhumanov A., Tyystjärvi T., Gorelova O., Baulina O., Chivkunova O., Solovchenko A., Rubin A. Photosynthetic hydrogen production as acclimation mechanism in nutrient-deprived Chlamydomonas // Algal Res. 2020. Vol. 49: 101951.
4. Antal T.K., Krendeleva T.E., Laurinavichene T.V., Makarova V.V., Ghirardi M.L., Rubin A.B., Tsygankov A.A, Seibert M. The dependence of algal H2 production on Photosystem II and O2 consumption activities in sulfur-deprived Chlamydomonas reinhardtii cells // Biochim. Biophys. Acta. 2003. Vol. 1607. N 2–3. P. 153–160.
5. Kosourov S., Seibert M., Ghirardi M.L. Effects of extracellular pH on the metabolic pathways in sulfur-deprived, H2-producing Chlamydomonas reinhardtii cultures // Plant Cell Physiol. 2003. Vol. 44. N 2. P. 146–155.
6. Gfeller R.P., Gibbs M. Fermentative metabolism of Chlamydomonas reinhardtii: I. analysis of fermentative products from starch in dark and light // Plant Physiol. 1984. Vol. 75. N 1. P. 212–218.
7. Atteia A., van Lis R., Gelius-Dietrich G., Adrait A., Garin J., Joyard J., Rolland N., Martin W. Pyruvate formatelyase and a novel route of eukaryotic ATP synthesis in Chlamydomonas mitochondria // J. Biol. Chem. 2006. Vol. 281. N 15. Р. 9909–9918.
8. Terashima M., Specht M., Naumann B., Hippler M. Characterizing the anaerobic response of Chlamydomonasreinhardtii by quantitative proteomics // Mol. Cell Proteomics. 2010. Vol. 9. N 7. P. 1514–1532.
9. Philipps G., Krawietz D., Hemschemeier A., Happe T. A pyruvate formate lyase (PFL1) deficient Chlamydomonas reinhardtii strain provides evidence for a link between fermentation and hydrogen evolution in green algae // Plant J. 2011. Vol. 66. N 2. P. 330–340.
10. Catalanotti C., Dubini A., Subramanian V., Yang W., Magneschi L., Mus F., Seibert M., Posewitz M.C., Grossman A.R. Altered fermentative metabolism in Chlamydomonas reinhardtii mutants lacking pyruvate formate lyase and both pyruvate formate lyase and alcohol dehydrogenase // Plant Cell. 2012. Vol. 24. N 2. P. 692–707.
11. Burgess S.J., Tredwell G., Molnàr A., Bundy J.G., Nixon P.J. Artificial microRNA-mediated knockdown of pyruvate formate lyase (PFL1) provides evidence for an active 3-hydroxybutyrate production pathway in the green alga Chlamydomonas reinhardtii // J. Biotechnol. 2012. Vol. 162. N 1. P. 57–66.
12. Laurinavichene T., Tolstygina I., Tsygankov A. The effect of light intensity on hydrogen production by sulfur-deprived Chlamydomonas reinhardtii // J. Biotechnol. 2004. Vol. 114. N 1–2. P. 143–151.
13. Lichtenthaler H.K. Chlorophylls and carotenoids: pigments of photosynthetic biomembranes // Methods Enzymol, vol. 148 / Eds. R. Douce and L. Packer. N.Y.: Press Inc., 1987. P. 350–382.
14. Shreiber U., Hormann H., Neubauer C., Klughammer C. Assessment of photosystem II photochemical quantum yield by chlorophyll fluorescence quenching analysis // Plant Physiol. 1995. Vol. 22. N 2. P. 209-220.
15. Hemschemeier A., Fouchard S., Cournac L., Peltier G., Happe T. Hydrogen production by Chlamydomonas reinhardtii: an elaborate interplay of electron sources and sinks // Planta. 2008. Vol. 227. N 2. P. 397-407.
16. Volgusheva A., Styring S., Mamedov F. Increased photosystem II stability promotes H2 production in sulfurdeprived Chlamydomonas reinhardtii // Proc. Natl. Acad. Sci. U.S.A. 2013. Vol. 110. N 18. P. 7223–7228.
Review
For citations:
Volgusheva A.A., Petrova E.V., Kukarskikh G.P., Dubini A., Antal T.K. Influence of fermentation reactions on continuous hydrogen photoproduction by microalga Chlamydomonas reinhardtii under sulfur deficiency. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2022;77(1):29-36. (In Russ.)