Research article Special Issues

Novel thermostable clostridial strains through protoplast fusion for enhanced biobutanol production at higher temperature—preliminary study

  • Received: 08 October 2015 Accepted: 24 December 2015 Published: 12 January 2016
  • The objective of this study is to improve the thermal stability of clostridium strains for enhanced biobutanol production. Thermostable clostridia species were developed through protoplast fusion between mesophilic clostridial species (i.e., Clostridium beijerinckii and Clostridium acetobutylicum) and thermophilic clostridial species (i.e., Clostridium thermocellum). Production of biobutanol was examined in the present preliminary study using the clostridium strains and their protoplast fusants using sugar mixture with composition identical to that of wheat straw acid hydrolysate. Maximum biobutanol production of 9.4 g/L was achieved by a fused strain at 45 °C with total sugar consumption of 66% compared to that at 35 °C (i.e., 8.4 g/L production and 64% total sugar consumption). Glucose and xylose uptake rates were generally higher compared to all other individual sugars in the feedstock. In general, average cell concentrations were in close proximity for all parenting and fused strains at 35 °C; i.e., in the range of 5.12 × 107 to 5.49 × 107 cells/mL. Average cell concentration of fusants between the mesophilic clostridial species and the thermophilic clostridial species slightly increased to ~ 5.62 × 107 cells/mL at a higher temperature of 45 °C. These results, in addition to the ones obtained for the butanol production, demonstrate enhanced thermal stability of both fusants at a higher temperature (45 °C).

    Citation: Muhammad Ferhan, Yaser Dahman. Novel thermostable clostridial strains through protoplast fusion for enhanced biobutanol production at higher temperature—preliminary study[J]. AIMS Energy, 2016, 4(1): 22-36. doi: 10.3934/energy.2016.1.22

    Related Papers:

  • The objective of this study is to improve the thermal stability of clostridium strains for enhanced biobutanol production. Thermostable clostridia species were developed through protoplast fusion between mesophilic clostridial species (i.e., Clostridium beijerinckii and Clostridium acetobutylicum) and thermophilic clostridial species (i.e., Clostridium thermocellum). Production of biobutanol was examined in the present preliminary study using the clostridium strains and their protoplast fusants using sugar mixture with composition identical to that of wheat straw acid hydrolysate. Maximum biobutanol production of 9.4 g/L was achieved by a fused strain at 45 °C with total sugar consumption of 66% compared to that at 35 °C (i.e., 8.4 g/L production and 64% total sugar consumption). Glucose and xylose uptake rates were generally higher compared to all other individual sugars in the feedstock. In general, average cell concentrations were in close proximity for all parenting and fused strains at 35 °C; i.e., in the range of 5.12 × 107 to 5.49 × 107 cells/mL. Average cell concentration of fusants between the mesophilic clostridial species and the thermophilic clostridial species slightly increased to ~ 5.62 × 107 cells/mL at a higher temperature of 45 °C. These results, in addition to the ones obtained for the butanol production, demonstrate enhanced thermal stability of both fusants at a higher temperature (45 °C).


    加载中
    [1] Formanek J, Mackie R, Blaschek HP (1997) Enhanced butanol production by Clostridium beijerinckii BA101 grown in semi-defined P2 medium containing 6 percent maltodextrin or glucose. Appl Environ Microbiol 63: 2306-2310.
    [2] Dahman Y (2012) Sustainable Biobutanol and Working towards the Green Gasoline of the Future. Ferment Technol 1: e111.
    [3] Durre P (1998) New insights and novel developments in clostridial acetone/butanol/isopropanol fermentation. Appl Microbiol Biotechnol 49: 639-648. doi: 10.1007/s002530051226
    [4] Yoo YG, Chao DY (1993) In: Genetics and breeding of edible mushrooms, Chang ST,  Buswell JA, and Miles PG (Eds.), Gordon and Breach Science Publishers, pp. 157-192.
    [5] Cocconcelli PS, Morelli L, Vescovo M, et al. (1986) Intergeneric protoplast fusion in lactic acid bacteria. FEMS Microbiol Lett 35: 211-214. doi: 10.1111/j.1574-6968.1986.tb01529.x
    [6] Lin FL, Sperle K, Sternberg N (1984) Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process. Mol Cell Biol 4: 1020-1034.
    [7] Zhao J, Chang ST (1995) Intraspecific hybridization between Coprinus cinereus and Schizophyllum commune by PEG—induced protoplast fusion and electro-fusion. World J Microbiol Biotechnol 11: 585-590. doi: 10.1007/BF00286378
    [8] Lynd LR, van Zyl WH, McBride, et al. (2005) Consolidated bioprocessing of cellulosic biomass: an update. Curr Opin Biotechnol 16: 577-583. doi: 10.1016/j.copbio.2005.08.009
    [9] Paredes CJ, Alsaker KV, Papoutsakis, et al. (2005) A comparative genomic view of clostridial differentiation and physiology. Nature Rev Microbiol 3: 969-978. doi: 10.1038/nrmicro1288
    [10] Johnson JL, Toth J, Santiwatanakul, et al. (1997) Cultures of Clostridium acetobutylicum from various collections comprise Clostridium acetobutylicum. Clostridium beijerinckii, and two other distinct types based on DNA-DNA reassociation. Int J Syst Bacteriol 47: 420-424.
    [11] Jones DT, Woods, et al. (1986) Acetone-butanol fermentation revisited. Microbiol Rev 50: 484-524.
    [12] Ahmed I, Ross RA, Mathur VK, et al. (1988) Growth rate dependence of solventogenesis and solvents produced by Clostridium beijerinckii. Appl Microbiol Biotechnol 28: 182-187.
    [13] Jones JDG, Dunsmuir P, Bedbrook J (1985) High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J 4: 2411-2418.
    [14] Dahman Y, Jayasuriya KE, Kallis M (2010) Potential of Biocellulose Nanofibers Production from Agricultural Renewable Resources: Preliminary Study. Appl Biochem Biotechnol 162: 1647-1659.
    [15] Thirmal C, Dahman Y (2012) Comparisons of existing pretreatment, saccharification, and fermentation processes for butanol production from agricultural residues. Can J Chem Eng 90: 745-761. doi: 10.1002/cjce.20601
    [16] Ezeji T, Qureshi N, Blaschek HP (2007) Butanol production from agricultural residues: impact of degradation products on Clostridium beijerinckii growth and butanol fermentation. Biotechnol Bioeng 97: 1460-1469. doi: 10.1002/bit.21373
    [17] Demain AL (2009) Biosolutions to the energy problem. J Ind Microbiol Biotechnol 36: 319-332. doi: 10.1007/s10295-008-0521-8
    [18] Ljungdahl LG (2009) A life with acetogens, thermophiles, and cellulolytic anaerobes. Annu Rev Microbiol 63: 1-25. doi: 10.1146/annurev.micro.091208.073617
    [19] Qureshi N, Saha BC, Cotta MA (2007) Butanol production from wheat straw hydrolysate using Clostridium beijerinckii. Bioprocess Biosyst Eng 30: 419-427.
    [20] Qureshi N, Saha BC, Hector RE, et al. (2008) Removal of fermentation inhibitors from alkaline peroxide pretreated and enzymatically hydrolyzed wheat straw: production of butanol from hydrolysate using Clostridium beijerinckii in batch reactors. Biomass Bioenerg 32: 1353-1358. doi: 10.1016/j.biombioe.2008.04.009
    [21] Qureshi N, Li XL, Hughes S, et al. (2006) Butanol production from corn fiber xylan using Clostridium acetobutylicum. Biotechnol Prog 22: 673-680. doi: 10.1021/bp050360w
    [22] Syed K, Dahman Y (2015) Novel clostridial fusants in comparison with co-cultured counterpart species for enhanced production of biobutanol using green renewable and sustainable feedstock. Bioprocess Biosyst Eng 11: 2249-2262.
    [23] Begum S, Dahman Y (2015) Enhanced biobutanol production using novel clostridial fusants in simultaneous saccharification and fermentation of green renewable agriculture residues. Biofuels, Bioprod Biorefin 5: 529-544.
    [24] MacFaddin JF (1985) Media for isolation-cultivation-identification-maintenance of medical bacteria, Vol. 1. William & Wilkins, Baltimore, Md.
    [25] Murray PR, Baron EJ, Pfaller MA, et al. (1995) Manual of Clinical Microbiology 6th Ed. American Society for Microbiology, Washington, DC.
    [26] Erick A, Madia JA, Demain AL (1981) Chemically defined Minimal Medium for growth of the anaerobic cellulolytic thermophile Clostridium thermocellum. Appl Environ Microbiol 41: 1060-1062.
    [27] O’Brien RW, Morris JG (1971) Oxygen and the growth and metabolism of Clostridium acetobutylicum. J Gen Microbiol 68: 307-318.
    [28] Allock ER, Reid SJ, Jones DT, et al. (1982) Clostridium acetobutylicum Protoplast Formation and Regeneration. Appl Environ Microbiol 43: 719-721.
    [29] Kong Q, He GQ, Chen F, et al. (2006) Studies on a kinetic model for butyric acid bioproduction by Clostridium butyricum. Lett Appl Microbiol 43: 71-77.
    [30] Hewitt CJ, Nebe Von-Caron G (2001) An industrial application of multiparameter flow cytometry: assessment of cell physiological state and its application to the study of microbial fermentations. Cytometry 44: 179-187.
    [31] Al-Abdallah W, Dahman Y (2013) Production of green biocellulose nanofibers by Gluconacetobacter xylinus through utilizing the renewable resources of agriculture residues. Bioprocess Biosyst Eng 36:1735-1743.
    [32] Heefner DL, Squires CH, Evans RJ, et al. (1984) Transformation of Clostridium perfringens. J. Bacteriol 159: 460-464.
    [33] Suto M, Tomita F (2001) Induction and catabolite repression mechanisms of cellulase in fungi. J Biosci Bioeng 92: 305-311.
    [34] van den Burg B, Vriend G, Veltman OR, et al. (1998) Engineering an enzyme to resist boiling. Proc Natl Acad Sci USA 95: 2056-2060. doi: 10.1073/pnas.95.5.2056
    [35] Romaniec M, Fauth U, Kobayashi T, et al. (1992) Purification and characterization of a new endoglucanase from Clostridium thermocellum. Biochem J 283: 69-73. doi: 10.1042/bj2830069
  • Reader Comments
  • © 2016 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Metrics

Article views(2287) PDF downloads(1036) Cited by(1)

Article outline

Figures and Tables

Figures(5)  /  Tables(4)

Other Articles By Authors

/

DownLoad:  Full-Size Img  PowerPoint
Return
Return

Catalog