| Peer-Reviewed

Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01

Received: 7 May 2022    Accepted: 6 June 2022    Published: 9 June 2022
Views:       Downloads:
Abstract

Hydrogen production yield of photosynthetic bacteria is low, which limits the development of the photo-fermentation. In order to solve this problem, We're trying to over-expression [FeFe]-hydrogenase in photosynthetic bacteria to improve its hydrogen production yield.[FeFe]-hydrogenase, from Rhodopseudomonas palustris CGA009 (RpHydC), Rhodospirillum rubrum ATCC 11170 (RrhydA), Clostridium acetobutylicum ATCC 824(CahydA) are selected as the research object. We construct the over-expression plasmid of pCG2,pAD2 and pCA2,which containing RphydC, RrhydA and CahydA DNA fragment, respectively. The hydG, hydE and hydF are amplified and cloned in pBBR1mcs-2 to form pEFG2. The strains A036 containing pCA2 and pEFG2, A037 containing pCG2 and pEFG2, A038 containing pAD2 and pEFG2.The hydrogen yield of A036, A037 and A038 are 1.38 mol H2/mol-glucose,1.34 mol H2/mol-glucose and 1.23 mol H2/mol-glucose,which compared with wild type strain increased by 29.71%, 25.52% and 15.07%, respectively.In this study, we successful implementation of the [FeFe]-hydrogenase heterologous expression in Rhodobacter sphaeroides HY01 by the test analysis of hydrogen production and RT-PCR. Otherwise, [FeFe]-hydrogenase,encoded by RphydC, RrhydA and CahydA are confirmed that their maturation was strictly dependent on co-expression of hydG, hydE, and hydF.

Published in Science Discovery (Volume 10, Issue 3)
DOI 10.11648/j.sd.20221003.25
Page(s) 173-180
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Heterologous Expression, [FeFe]-hydrogenase, Hydrogen Yield

References
[1] Balat H, Kirtay E. Hydrogen from biomass - Present scenario and future prospects [J]. Int J Hydrogen Energ, 2010, 35 (14): 7416-26.
[2] Shimura K, Yoshida H. Heterogeneous photocatalytic hydrogen production from water and biomass derivatives [J]. Energ Environ Sci, 2011, 4 (7): 2467-81.
[3] Zhao X, Xing DF, Zhang L, Ren NQ. Characterization and overexpression of a [FeFe]-hydrogenase gene of a novel hydrogen-producing bacterium Ethanoligenens harbinense [J]. Int J Hydrogen Energ, 2010, 35 (18): 9598-602.
[4] Demirbas A. Progress and recent trends in biofuels [J]. Prog Energ Combust, 2007, 33 (1): 1-18.
[5] John RP, Anisha GS, Nampoothiri KM, Pandey A. Micro and macroalgal biomass: A renewable source for bioethanol [J]. Bioresource Technol, 2011, 102 (1): 186-93.
[6] Holladay JD, Hu J, King DL, Wang Y. An overview of hydrogen production technologies [J]. Catal Today, 2009, 139 (4): 244-60.
[7] Xia A, Cheng J, Ding LK, Lin RC, Huang R, Zhou JH, et al. Improvement of the energy conversion efficiency of Chlorella pyrenoidosa biomass by a three-stage process comprising dark fermentation, photofermentation, and methanogenesis [J]. Bioresource Technol, 2013, 146: 436-43.
[8] Akutsu Y, Li YY, Harada H, Yu HQ. Effects of temperature and substrate concentration on biological hydrogen production from starch [J]. Int J Hydrogen Energ, 2009, 34 (6): 2558-66.
[9] van Veen JAR, Sie ST. Fuel processing technology special issue: Deep hydrodesulfurization of diesel fuel [J]. Fuel Process Technol, 1999, 61 (1-2): 1-4.
[10] Chandrasekhar K, Lee YJ, Lee DW. Biohydrogen Production: Strategies to Improve Process Efficiency through Microbial Routes [J]. Int J Mol Sci, 2015, 16 (4): 8266-93.
[11] Ren HY, Liu BF, Xie GJ, Zhao L, Ren NQ. Carrier modification and its application in continuous photo-hydrogen production using anaerobic fluidized bed photo-reactor [J]. Gcb Bioenergy, 2014, 6 (5): 599-605.
[12] Burrows EH, Wong WK, Fern X, Chaplen FWR, Ely RL. Optimization of pH and Nitrogen for Enhanced Hydrogen Production by Synechocystis sp PCC 6803 via Statistical and Machine Learning Methods [J]. Biotechnol Progr, 2009, 25 (4): 1009-17.
[13] Fan YT, Li CL, Lay JJ, Hou HW, Zhang GS. Optimization of initial substrate and pH levels for germination of sporing hydrogen-producing anaerobes in cow dung compost [J]. Bioresource Technol, 2004, 91 (2): 189-93.
[14] Hiligsmann S, Beckers L, Masset J, Hamilton C, Thonart P. Improvement of fermentative biohydrogen production by Clostridium butyricum CWBI1009 in sequenced-batch, horizontal fixed bed and biodisc-like anaerobic reactors with biomass retention [J]. Int J Hydrogen Energ, 2014, 39 (13): 6899-911.
[15] Chang JS, Lee KS, Lin PJ. Biohydrogen production with fixed-bed bioreactors [J]. Int J Hydrogen Energ, 2002, 27 (11-12): 1167-74.
[16] Gomez X, Fernandez C, Fierro J, Sanchez ME, Escapa A, Moran A. Hydrogen production: Two stage processes for waste degradation [J]. Bioresource Technol, 2011, 102 (18): 8621-27.
[17] Eroglu E, Eroglu I, Gunduz U, Turker L, Yucel M. Biological hydrogen production from olive mill wastewater with two-stage processes [J]. Int J Hydrogen Energ, 2006, 31 (11): 1527-35.
[18] Rajhi H, Conthe M, Puyol D, Diaz E, Sanz JL. Dark fermentation: isolation and characterization of hydrogen-producing strains from sludges [J]. Int Microbiol, 2013, 16 (1): 53-62.
[19] Laocharoen S, Reungsang A. Isolation, characterization and optimization of photo-hydrogen production conditions by newly isolated Rhodobacter sphaeroides KKU-PS5 [J]. Int J Hydrogen Energ, 2014, 39 (21): 10870-82.
[20] Das D, Veziroglu TN. Advances in biological hydrogen production processes [J]. Int J Hydrogen Energ, 2008, 33 (21): 6046-57.
[21] Ren NQ, Wang AJ, Cao GL, Xu JF, Gao LF. Bioconversion of lignocellulosic biomass to hydrogen: Potential and challenges [J]. Biotechnol Adv, 2009, 27 (6): 1051-60.
[22] Abo-Hashesh M, Sabourin-Prouost G, Hallenbeck PC. RrHydA is inactive when overexpressed in Rhodospirillum rubrum but can be matured in Escherichia coli [J]. Int J Hydrogen Energ, 2013, 38 (26): 11233-40.
[23] Liu T, Li XF, Zhou ZH. Improvement of hydrogen yield by hupR gene knock-out and nifA gene overexpression in Rhodobacter sphaeroides 6016 [J]. Int J Hydrogen Energ, 2010, 35 (18): 9603-10.
[24] Kars G, Gunduz U, Rakhely G, Yucel M, Eroglu I, Kovacs KL. Improved hydrogen production by uptake hydrogenase deficient mutant strain of Rhodobacter sphaeroides OU001 [J]. Int J Hydrogen Energ, 2008, 33 (12): 3056-60.
[25] Wang JL, Gray KA, Daldal F, Rousseau DL. The Cbb3-Type Cytochrome-C-Oxidase from Rhodobacter-Capsulatus Contains a Unique Active-Site [J]. J Am Chem Soc, 1995, 117 (36): 9363-64.
[26] Brotosudarmo THP, Collins AM, Gall A, Roszak AW, Gardiner AT, Blankenship RE, et al. The light intensity under which cells are grown controls the type of peripheral light-harvesting complexes that are assembled in a purple photosynthetic bacterium [J]. Biochem J, 2011, 440: 51-61.
[27] Paoli GC, Morgan NS, Tabita FR, Shively JM. Expression of the cbbLcbbS and cbbM genes and distinct organization of the cbb Calvin cycle structural genes of Rhodobacter capsulatus [J]. Arch Microbiol, 1995, 164 (6): 396-405.
[28] Akhtar MK, Jones PR. Engineering of a synthetic hydF-hydE-hydG-hydA operon for biohydrogen production [J]. Anal Biochem, 2008, 373 (1): 170-72.
[29] Kim EJ, Lee MK, Kim MS, Lee JK. Molecular hydrogen production by nitrogenase of Rhodobacter sphaeroides and by Fe-only hydrogenase of Rhodospirillum rubrum [J]. Int J Hydrogen Energ, 2008, 33 (5): 1516-21.
[30] Yang HH, Zhang J, Wang XQ, Feng JT, Yan W, Guo LJ. A newly isolated Rhodobacter sphaeroides HY01 with high hydrogen production performance [J]. Int J Hydrogen Energ, 2014, 39 (19): 10051-60.
[31] An D, Li Q, Wang XQ, Yang HH, Guo LJ. Characterization on hydrogen production performance of a newly isolated Clostridium beijerinckii YA001 using xylose [J]. Int J Hydrogen Energ, 2014, 39 (35): 19928-36.
[32] Ma C, Guo L, Yang H. Improved photo – Hydrogen production by transposon mutant of Rhodobacter capsulatus with reduced pigment [J]. Int J Hydrogen Energ, 2012, 37 (17): 12229-33.
[33] Ma C, Yang H, Zhang Y, Guo L. Disruption of multidrug resistance protein gene of Rhodobacter capsulatus results in improved photoheterotrophic hydrogen production [J]. Int J Hydrogen Energ, 2013, 38 (29): 13031-37.
[34] Posewitz MC, King PW, Smolinski SL, Zhang LP, Seibert M, Ghirardi ML. Discovery of two novel radical S-adenosylmethionine proteins required for the assembly of an active [Fe] hydrogenase[J]. J Biol Chem, 2004, 279 (24): 25711-20.
Cite This Article
  • APA Style

    An Dan, Song Zi-lun, Su Zhen-hua, Wei Hao-wen, Xiang Ming-rui, et al. (2022). Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01. Science Discovery, 10(3), 173-180. https://doi.org/10.11648/j.sd.20221003.25

    Copy | Download

    ACS Style

    An Dan; Song Zi-lun; Su Zhen-hua; Wei Hao-wen; Xiang Ming-rui, et al. Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01. Sci. Discov. 2022, 10(3), 173-180. doi: 10.11648/j.sd.20221003.25

    Copy | Download

    AMA Style

    An Dan, Song Zi-lun, Su Zhen-hua, Wei Hao-wen, Xiang Ming-rui, et al. Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01. Sci Discov. 2022;10(3):173-180. doi: 10.11648/j.sd.20221003.25

    Copy | Download

  • @article{10.11648/j.sd.20221003.25,
      author = {An Dan and Song Zi-lun and Su Zhen-hua and Wei Hao-wen and Xiang Ming-rui and Yin Yue},
      title = {Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01},
      journal = {Science Discovery},
      volume = {10},
      number = {3},
      pages = {173-180},
      doi = {10.11648/j.sd.20221003.25},
      url = {https://doi.org/10.11648/j.sd.20221003.25},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.sd.20221003.25},
      abstract = {Hydrogen production yield of photosynthetic bacteria is low, which limits the development of the photo-fermentation. In order to solve this problem, We're trying to over-expression [FeFe]-hydrogenase in photosynthetic bacteria to improve its hydrogen production yield.[FeFe]-hydrogenase, from Rhodopseudomonas palustris CGA009 (RpHydC), Rhodospirillum rubrum ATCC 11170 (RrhydA), Clostridium acetobutylicum ATCC 824(CahydA) are selected as the research object. We construct the over-expression plasmid of pCG2,pAD2 and pCA2,which containing RphydC, RrhydA and CahydA DNA fragment, respectively. The hydG, hydE and hydF are amplified and cloned in pBBR1mcs-2 to form pEFG2. The strains A036 containing pCA2 and pEFG2, A037 containing pCG2 and pEFG2, A038 containing pAD2 and pEFG2.The hydrogen yield of A036, A037 and A038 are 1.38 mol H2/mol-glucose,1.34 mol H2/mol-glucose and 1.23 mol H2/mol-glucose,which compared with wild type strain increased by 29.71%, 25.52% and 15.07%, respectively.In this study, we successful implementation of the [FeFe]-hydrogenase heterologous expression in Rhodobacter sphaeroides HY01 by the test analysis of hydrogen production and RT-PCR. Otherwise, [FeFe]-hydrogenase,encoded by RphydC, RrhydA and CahydA are confirmed that their maturation was strictly dependent on co-expression of hydG, hydE, and hydF.},
     year = {2022}
    }
    

    Copy | Download

  • TY  - JOUR
    T1  - Improved Hydrogen Production by Overexpression of RphydC, RrhydA, CahydA in Rhodobacter sphaeroides HY01
    AU  - An Dan
    AU  - Song Zi-lun
    AU  - Su Zhen-hua
    AU  - Wei Hao-wen
    AU  - Xiang Ming-rui
    AU  - Yin Yue
    Y1  - 2022/06/09
    PY  - 2022
    N1  - https://doi.org/10.11648/j.sd.20221003.25
    DO  - 10.11648/j.sd.20221003.25
    T2  - Science Discovery
    JF  - Science Discovery
    JO  - Science Discovery
    SP  - 173
    EP  - 180
    PB  - Science Publishing Group
    SN  - 2331-0650
    UR  - https://doi.org/10.11648/j.sd.20221003.25
    AB  - Hydrogen production yield of photosynthetic bacteria is low, which limits the development of the photo-fermentation. In order to solve this problem, We're trying to over-expression [FeFe]-hydrogenase in photosynthetic bacteria to improve its hydrogen production yield.[FeFe]-hydrogenase, from Rhodopseudomonas palustris CGA009 (RpHydC), Rhodospirillum rubrum ATCC 11170 (RrhydA), Clostridium acetobutylicum ATCC 824(CahydA) are selected as the research object. We construct the over-expression plasmid of pCG2,pAD2 and pCA2,which containing RphydC, RrhydA and CahydA DNA fragment, respectively. The hydG, hydE and hydF are amplified and cloned in pBBR1mcs-2 to form pEFG2. The strains A036 containing pCA2 and pEFG2, A037 containing pCG2 and pEFG2, A038 containing pAD2 and pEFG2.The hydrogen yield of A036, A037 and A038 are 1.38 mol H2/mol-glucose,1.34 mol H2/mol-glucose and 1.23 mol H2/mol-glucose,which compared with wild type strain increased by 29.71%, 25.52% and 15.07%, respectively.In this study, we successful implementation of the [FeFe]-hydrogenase heterologous expression in Rhodobacter sphaeroides HY01 by the test analysis of hydrogen production and RT-PCR. Otherwise, [FeFe]-hydrogenase,encoded by RphydC, RrhydA and CahydA are confirmed that their maturation was strictly dependent on co-expression of hydG, hydE, and hydF.
    VL  - 10
    IS  - 3
    ER  - 

    Copy | Download

Author Information
  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • School of Enviromental Science and Engineering, Shaanxi University of Science & Technology, Xi’an, China

  • Sections