Skip to main content
SpringerLink
Log in

Thermostable Enzymes in Lignocellulose Hydrolysis

  • Chapter
  • First Online:
Biofuels

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 108))

Abstract

Thermostable enzymes offer potential benefits in the hydrolysis of lignocellulosic substrates; higher specific activity decreasing the amount of enzymes, enhanced stability allowing improved hydrolysis performance and increased flexibility with respect to process configurations, all leading to improvement of the overall economy of the process. New thermostable cellulase mixtures were composed of cloned fungal enzymes for hydrolysis experiments. Three thermostable cellulases, identified as the most promising enzymes in their categories (cellobiohydrolase, endoglucanase and β-glucosidase), were cloned and produced in Trichoderma reesei and mixed to compose a novel mixture of thermostable cellulases. Thermostable xylanase was added to enzyme preparations used on substrates containing residual hemicellulose. The new optimised thermostable enzyme mixtures were evaluated in high temperature hydrolysis experiments on technical steam pretreated raw materials: spruce and corn stover. The hydrolysis temperature could be increased by about 10–15 °C, as compared with present commercial Trichoderma enzymes. The same degree of hydrolysis, about 90% of theoretical, measured as individual sugars, could be obtained with the thermostable enzymes at 60 °C as with the commercial enzymes at 45 °C. Clearly more efficient hydrolysis per assayed FPU unit or per amount of cellobiohydrolase I protein used was obtained. The maximum FPU activity of the novel enzyme mixture was about 25% higher at the optimum temperature at 65 °C, as compared with the highest activity of the commercial reference enzyme at 60 °C. The results provide a promising basis to produce and formulate improved enzyme products. These products can have high temperature stability in process conditions in the range of 55–60 °C (with present industrial products at 45–50 °C) and clearly improved specific activity, essentially decreasing the protein dosage required for an efficient hydrolysis of lignocellulosic substrates. New types of process configurations based on thermostable enzymes are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 259.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 329.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 329.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Abrha B, Gashe BA (1992) Cellulase production and activity in a species of Cladosporium. World J Microbiol Biotech 8:164–166

    Article  CAS  Google Scholar 

  2. Barnett C Berka R, Fowler T (1991) Cloning and amplification of the gene encoding an extracellular β-glucose from Trichoderma reesei: evidence for improved rates of saccharification of cellulosic substrates. Biotechnology 9:562–567

    Article  Google Scholar 

  3. Barr B Hsieh, Ganem B, Wilson BD (1996) Identification of two functionally different classes of exocellulases. Biochemistry 35:586–592

    Article  Google Scholar 

  4. Bailey MJ, Biely P, Poutanen K (1992) Interlaboratory testing methods for assay of xylanase activity. J Biotechnol 23:257–270

    Article  CAS  Google Scholar 

  5. Bailey M, Nevalainen H (1981) Induction, isolation and testing of stable Trichoderma reesei mutants with improved production of solubilizing cellulase. Enzyme Microb Technol 3:153–157

    Article  CAS  Google Scholar 

  6. Bailey MJ, Tähtiharju J (2003) Efficient cellulase production by Trichoderma reesei in continuous cultivation on lactose medium with a computer-controlled feeding strategy. Appl Microbiol Biotechnol 62:156–162

    Article  CAS  Google Scholar 

  7. Bergquist P, Gibbs M, Morris D, Te'o V, Jsaul D, Morgan H (1999) Molecular diversity of thermophilic cellulolytic and hemicellulolytic bacteria. FEMS Microbiol Ecol 28:99–110

    Article  CAS  Google Scholar 

  8. Bergquist PL, Te'o VSJ, Gibbs MD, Curah NC, Nevalainen KMH (2004) Recombinant enzymes from thermophilic micro-organisms expressed in fungal hosts. Biochem Soc Trans 32(2):293–297

    Article  CAS  Google Scholar 

  9. Berlin A, Gilkes N, Kilburn D, Bura R, Markov A, Skomarovsky A, Okunev O, Gusakov A, Maximenko V, Gregg D, Sinitsyn A, Saddler J (2005) Evaluation of novel fungal cellulase preparations for ability to hydrolyze softwood substrates – evidence for the role of accessory enzymes. Enzyme Microb Technol 37:175–184

    Article  CAS  Google Scholar 

  10. Bernfeld P (1955) Amylases α and β. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 1. Academic, New York, pp 149–158

    Google Scholar 

  11. Bok JD, Yernool DA, Eveleigh DE (1998) Purification, characterization, and molecular analysis of thermostable cellulases CelA and CelB from Thermotoga neapolitana. Appl Environ Microbiol 64:4774–4781

    CAS  Google Scholar 

  12. Bronnenmeier K, Rücknagel K, Staudenbauer W (1991) Purification and properties of a novel type exo-1,4-β-glucanase (Avicelase II) from the cellulolytic thermophile Clostridium stercorarium. Eur J Biochem 200:379–385

    Article  CAS  Google Scholar 

  13. Bronnenmeier K, Staudenbauer W (1990) Cellulose hydrolysis by a highly thermostable endo-1,4-glucanase (Avicelase I) from Clostridium stercorarium. Enzyme Microbial Technol 12:431–436

    Article  CAS  Google Scholar 

  14. Bronnenmeier K, Kern A, Liebl W, Staudenbauer W (1995) Purification of Thermotoga maritima enzymes for the degradation of cellulosic materials. Appl Environ Microbiol 61:1339–1407

    Google Scholar 

  15. Coughlan M, McHale A (1988) Purification of β-d-glucoside glucohydrolases of Talaromyces emersonii. Methods Enzymol 160:437–443

    Article  CAS  Google Scholar 

  16. Coutinho PM, Henrissat B (1999) Carbohydrate active enzymes. Database available at http://www.cazy.org , last visited: 10 May 2007

  17. Demain A, Newcomb M, Wu JHD (2005) Cellulase, clostridia and ethanol. Microb Molec Biol Rev 69:124–154

    Article  CAS  Google Scholar 

  18. Ding S-Y (2006) Thermotolerant cellulase. Industrial Bioproc 28(7):3–4

    CAS  Google Scholar 

  19. Divne C, Ståhlberg J, Teeri T, Jones T (1998) High-resolution crystal structures reveal how a cellulose chain is bound in the 50 Å long tunnel of cellobiohydrolase I from Trichoderma reesei. J Mol Biol 275:309–325

    Article  CAS  Google Scholar 

  20. Eklund R, Galbe M, Zacchi G (1990) Optimization of temperature and enzyme concentration in the enzymatic saccharification of steam pretreated willow. Enzyme Microb Technol 12:225–228

    Article  CAS  Google Scholar 

  21. Evans B, Gilman A, Cordray K, Woodward J (2000) Mechanism of substrate hydrolysis by a thermophilic endoglucanase from Thermotoga maritima. Biotechnol Lett 22:735–740

    Article  CAS  Google Scholar 

  22. Fauth U, Romaniec M, Kobayashi T, Demain A (1991) Purification and characterization of endoglucanase Ss from Clostridium thermocellum. Biochem J 279:67–73

    CAS  Google Scholar 

  23. Feldman KA, Lovett JS, Tsao GT (1988) Isolation of cellulase enzymes from the thermophilic fungus Thermoascus aurantiacus and regulation of enzyme production. Enzyme Microb Technol 10:262–272

    Article  CAS  Google Scholar 

  24. Foreman PK, Brown D, Dankmeyer L, Dean R, Diener S, Dunn-Coleman NS, Goedegebuur F, Houfek T, England G, Kelley A, Meerman H, Mitchell T, Mitchinson C, Olivares H, Teunissen P, Yao J, Ward M (2003) Transcriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei. J Biol Chem 278(34):31988–31997

    Article  Google Scholar 

  25. Fullbrook PD (1996) Practical limits and prospects (kinetics). In: Godfrey T, West S (eds) Industrial enzymology, 2nd edn. MacMillan, London, pp 508–509

    Google Scholar 

  26. Gomes I, Gomes J, Gomes D, Steiner W (2000) Simultaneous production of high activities of thermostable endoglucanase and β-d-glucosidase by the wild thermophilic fungus Thermoascus aurantiacus. Appl Microbiol Biotechnol 53:461–468

    Article  CAS  Google Scholar 

  27. Grassick A, Murray P, Thompson R, Collins C, Byrnes L, Birrane G, Higgins T, Tuohy M (2004) Three-dimensional structure of a thermostable native cellobiohydrolase, CBH IB, and molecular characterization of the cel7 gene from the filamentous fungus, Talaromyces emersonii. Eur J Biochem 271:4495–4506

    Article  CAS  Google Scholar 

  28. Grishutin S, Gusakov A, Markov A, Ustinov B, Semenova M, Sinitsyn A (2004) Specific xyloglucanases as a new class of polysaccharide-degrading enzymes. Biochim Biophys Acta 1674:268–281

    CAS  Google Scholar 

  29. Hakamada Y, Koike K, Yoshimatsu T, Mori H, Kobayashi T, Ito S (1997) Thermostable alkaline cellulase from an alkaliphilic isolate, Bacillus sp. KSM-S237. Extremophiles 1:151–156

    Article  CAS  Google Scholar 

  30. Harchand R, Singh S (1997) Characterization of cellulase complex of Streptomyces albaduncus; thermostable cellulase, cellobiohydrolase and beta-glucosidase characterization. J Basic Microbiol 37:93–103

    Article  CAS  Google Scholar 

  31. Henrissat B, Driquez H, Viet C, Schülein M (1985) Synergism of cellulases from Trichoderma reesei in degradation of cellulose. Bio/Technology 3:722–726

    Article  CAS  Google Scholar 

  32. Himmel M, Adney W, Tucker M, Grohmann K (1994) Thermostable purified endoglucanase from Acidothermus cellulolyticus ATCC 43068. US Patent 5275944

    Google Scholar 

  33. Hogsett DA, Ahn H-J, Bernardez TD, South CR, Lynd LR (1992) Direct microbial conversion. Prospects, progress and obstacles. Appl Biochem Biotechnol 34:527–541

    Article  Google Scholar 

  34. Hreggvidsson G, Kaiste E, Holst O, Eggertsson G, Palsdottir A, Kristjansson A (1996) An extremely thermostable cellulase from the thermophilic eubacterium Rhodothermus marinus. Appl Environ Microbiol 62:3047–3049

    CAS  Google Scholar 

  35. Ishihara M, Shinkichi T, Toyama S (1999) Disintegration of uncooked rice by carboxymethyl cellulase from Sporotrichum sp. HG-I. J Biosci Bioeng 87:249–251

    Article  CAS  Google Scholar 

  36. International Union of Pure and Applied Chemistry (IUPAC) (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268

    Google Scholar 

  37. Jang HD, Chen KS (2003) Production and characterization of thermostable cellulases from Streptomyces transformant T3-1. World J Microb Biotechnol 19:263–268

    Article  CAS  Google Scholar 

  38. Karlsson J, Momcilovic D, Wittgren B, Schülein M, Tjerneld F, Brinkman G (2002) Enzymatic degradation of carboxymethyl cellulose hydrolysed by the endoglucanases Cel5A, Cel7B and Cel45A from Humicola insolens and Cel7B, Cel12A and Cel45A core from Trichoderma reesei. J Biotechnol 63:32–40

    CAS  Google Scholar 

  39. Klyosov A (1990) Trends in biochemistry and enzymology of cellulose degradation. Biochemistry 29:10577–10585

    Article  CAS  Google Scholar 

  40. Lin J, Pillay B, Singh S (1999) Purification and biochemical characteristics of β-d-glucosidase from a thermophilic fungus, Thermomyces lanuginosus–SSBP. Biotechnol Appl Biochem 30:81–87

    CAS  Google Scholar 

  41. Lin S, Stutzenberger F (1995) Purification and characterization of the major beta-1,4-endoglucanase from Thermomonospora curvata. J Appl Bacteriol 79:447–53

    CAS  Google Scholar 

  42. Li D-C, Lu M, Li Y-A, Lu J (2003) Purification and characterization of an endocellulase from the thermophilic fungus Chaetomium thermophilum CT 2. Enzyme Microb Technol 33:932–937

    Article  CAS  Google Scholar 

  43. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the Folin phenol reagent. J Biol Chem 193:265–75

    CAS  Google Scholar 

  44. Macarron R, Acebal C, Castillon M, Claeyssens M (1996) Mannanase activity of endoglucanase III from Trichoderma reesei QM9414. Biochem Lett 18:599–602

    Article  CAS  Google Scholar 

  45. Maheshwari R, Bharadwaj G, Bhat M (2000) Thermophilic fungi: Their physiology and enzymes. Microbiol Mol Biol Rev 64:461–488

    Article  CAS  Google Scholar 

  46. Medve J, Ståhlherg J, Tjerneld F (1994) Adsorption and synergism of cellobiohydrolase I and II of Trichoderma reesei during hydrolysis of microcrystalline cellulose. Biotechnol Bioeng 44:1064–1073

    Article  CAS  Google Scholar 

  47. Miettinen-Oinonen A, Londesborough J, Vehmaanperä J, Haakana H, Mäntylä A, Lantto R, Elovainio M, Joutsjoki V, Paloheimo M, Suominen P (1996) New cellulases from fungi for use in pulp and textile processing and the genes encoding the enzymes. WO Patent no. 9714804

    Google Scholar 

  48. Millner PD (1977) Radial growth responses to temperature by 58 Chaetomium species and some taxonomic relationships. Mycologia 69:492–502

    Article  Google Scholar 

  49. Mooney C, Mansfield S, Tuohy M, Saddler J (1998) The effect of the initial pore volume and lignin content on the enzymatic hydrolysis of softwoods. Biores Technol 64:113–119

    Article  CAS  Google Scholar 

  50. Murray P, Grassick A, Laffey C, Cuffe M, Higgins T, Savage A, Planas A, Tuohy M (2001) Isolation and characterization of a thermostable endo-β-glucanase active on 1,3-1,4-β-d-glucans from the aerobic fungus Talaromyces emersonii CBS 814.70. Enzyme Microb Technol 29:90–98

    Article  CAS  Google Scholar 

  51. Nidetzky B, Claeyssens M (1994) Specific quantification of Trichoderma reesei cellulases in reconstituted mixtures and its application to cellulose-cellulose binding studies. Biotechnol Bioeng 44:961–966

    Article  CAS  Google Scholar 

  52. Nidetzky B, Hayn M, Macarron R, Steiner W (1993) Synergism of Trichoderma reesei cellulases while degrading different celluloses. Biotechnol Lett 151:71–76

    Article  Google Scholar 

  53. Nidetzky B, Steiner W, Hayn M, Claeyssens M (1994) Cellulose hydrolysis by the cellulases from Trichoderma reesei: A new model for synergistic interaction. Biochem J 298:705–710

    CAS  Google Scholar 

  54. Nigam P, Prabhu KA (1991) Influence of sugars on the activity of cellulose systems from two basidomycetes cultures. J Basic Microbiol 31:279–283

    Article  CAS  Google Scholar 

  55. Nutt A, Sild V, Petterson G, Johansson G (1998) Progress curves a mean for functional classification of cellulases. Eur J Biochem 258:200–206

    Article  CAS  Google Scholar 

  56. Palonen H, Viikari L (2004) Role of oxidative enzymatic treatments on enzymatic hydrolysis of softwood. Biotechnol Bioeng 86:550–557

    Article  CAS  Google Scholar 

  57. Park C, Kawaguchi T, Sumitani J, Arai M (2001) Purification and characterization of cellulases (CBH I and EGL 1) produced by thermophilic microorganism Streptomyces sp. M23. Appl Biol Sci 7(1):27–35

    CAS  Google Scholar 

  58. Parry N, Beever D, Owen E, Vandenbergh I, van Beeumen J, Bhat M (2001) Biochemical characterization and mechanism of action of a thermostable β-glucosidase purified from Thermoascus aurantiacus. Biochem J 353:117–127

    Article  CAS  Google Scholar 

  59. Parry N, Beever D, Owen E, Nerinckx W, Claeyssens M, Van Beeumen J, Bhat M (2002) Biochemical characterization and mode of action of a thermostable endoglucanase purified from Thermoascus aurantiacus. Arch Biochem Biophys 404:243–253

    Article  CAS  Google Scholar 

  60. Phillippidis GP, Smith TK, Wyman CE (1993) Study of the enzymatic hydrolysis of cellulose for production of fuel ethanol by the simultaneous saccharification and fermentation process. Biotechnol Bioeng 41:846–853

    Article  Google Scholar 

  61. Romaniec M, Fauth U, Kobayashi T, Huskisson N, Barker P, Demain A (1992) Purification and characterization of a new endoglucanase from Clostridium thermocellum. Biochem J 283:69–73

    CAS  Google Scholar 

  62. Rosgaard L, Pedersen S, Cherry JR, Harris P, Meyer AS (2006) Efficiency of new fungal cellulose systems in boosting enzymatic degradation of barley straw lignocellulose. Biotechnol Progr 22:493–498

    Article  CAS  Google Scholar 

  63. Roy S, Raha S, Dey S, Chakrabarty S (1990) Effect of temperature on the production and location of cellulose components in Myceliopthora thermophila D-14 (ATCC 48104). Enzyme Microb Technol 12:710–713

    Article  CAS  Google Scholar 

  64. Ruttersmith L, Daniel R (1991) Thermostable cellobiohydrolase from the thermophilic eubacterium Thermotoga sp. strain FjSS3-B.1: purification and properties. Biochem J 277:887–890

    CAS  Google Scholar 

  65. Saboto D, Nucci R, Rossi M, Gryczynski I, Gryczynski Z, Lakowicz J (1999) The β-glycosidase from the hyperthermophilic archaeon Sulfolobus solfataricus: enzyme activity and conformational dynamics at temperatures above 100 °C. Biophys Chem 81:23–31

    Article  Google Scholar 

  66. Saha B, Freer S, Bothast R (1994) Production, purification and properties of a thermostable β-glucosidase from a color variant strain of Aureobasidium pullulans. Appl Env Microbiol 60:3774–3780

    CAS  Google Scholar 

  67. Sakon J, Adney W, Himmel M, Thomas S, Karplus P (1996) Crystal structure of thermostable family 5 endoglucanase EI from Acidothermus cellulolyticus in complex with cellotetraose. Biochemistry 35:10648–10660

    Article  CAS  Google Scholar 

  68. Sarker M, Ilias M, Mozammel H (1998) Charaterization of xylanase and CMCase from Rhizomucor pusillus. Bangladesh J Microbiol 15(2):41–47

    Google Scholar 

  69. Sassner P, Galbe M, Zacchi G (2006) Bioethanol production based on simultaneous saccharification and fermentation of steam-pretreated salix at high dry-matter content. Enzyme Microb Technol 39:756–762

    Article  CAS  Google Scholar 

  70. Shepherd M, Tong C, Cole A (1981) Substrate specificity and mode of action of the cellulases from the thermophilic fungus Thermoascus aurantiacus. Biochem J 193:67–84

    CAS  Google Scholar 

  71. Shoemaker S, Watt K, Tsikovsky G, Cox R (1983) Characterization and properties of cellulases purified from Trichoderma reesei strain L27. Bio/Technology 1:687–690

    Article  CAS  Google Scholar 

  72. Stenberg K, Bollok M, Reczey K, Galbe M, Zacchi G (2000) Effect of substrate and cellulose concentration on simultaneous saccharification and fermentation of steam pretreated softwood for ethanol production. Biotechnol Bioeng 68:204–210

    Article  CAS  Google Scholar 

  73. Ståhlberg J (1991) Functional organization of cellulases from Trichoderma reesei. PhD Thesis, Uppsala University, Sweden

    Google Scholar 

  74. Takashima S, Nakamura A, Hidaka M, Masaki H, Uozumi T (1999) Molecular cloning and expression of the novel fungal β-glucosidase genes from Humicola grisea and Trichoderma reesei. J Biochem 125:728–736

    CAS  Google Scholar 

  75. Tenkanen M, Siika-aho M (2000) An α-glucuronidase of Schizophyllum commune acting on polymeric xylan. J Biotechnol 75:149–161

    Article  Google Scholar 

  76. Te'o V, Saul D, Bergquist P (1995) CelA, another gene coding for a multidomain cellulases from the extreme thermophile Caldocellum saccharolyticum. Appl Microbiol Biotechnol 43:291–296

    Article  Google Scholar 

  77. Tomme P, Heriben V, Claeyssens M (1990) Adsorption of two cellobiohydrolases from Trichoderma reesei to Avicel: Evidence for exo–exo synergism and possible “loose complex” formation. Biotechnol Lett 127:525–530

    Article  Google Scholar 

  78. Tuohy M, Walsh J, Murray P, Claeyssens M, Cuffe M, Savage A, Coughlan M (2002) Kinetic parameters and mode of action of the cellobiohydrolases produced by Talaromyces emersonii. Biochem Biophys Acta 1596:366–380

    Article  CAS  Google Scholar 

  79. Venturi L, Polizeli M, Terenzi H, Furriel R, Jorge J (2002) Extracellular β-d-glucosidase from Chaetomium thermophilum var. coprophilum: production, purification and some properties. J Basic Microbiol 42:55–66

    Article  CAS  Google Scholar 

  80. Väljamäe P, Sild V, Petterson G, Johansson G (1998) The initial kinetics of hydrolysis by cellobiohydrolase I and II is consistent with a cellulose surface – erosion model. Eur J Biochem 253:469–475

    Article  Google Scholar 

  81. Wright JD, Wyman CE, Grohmann K (1988) Simultaneous saccharification and fermentation of lignocellulose: process evaluation. Appl Biochem Biotechnol 18:75–90

    Article  CAS  Google Scholar 

  82. Zverlov V, Mahr S, Riedel K, Bronnenmeier K (1998) Properties and gene structure of a bifunctional cellulolytic enzyme (CelA) from the extreme thermophile Anaerocellum thermophilum with separate glycosyl hydrolase family 9 and 48 catalytic domains. Microbiology 143:3537–3542

    Article  Google Scholar 

  83. Öhgren K, Vehmaanperä J, Siika-aho M, Galbe M, Viikari L, Zacchi G (2007) High temperature enzymatic hydrolysis prior to simultaneous saccharification and fermentation of steam pretreated corn stover for ethanol production. Enzyme Microb Technol 40:607–613

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. University of Helsinki, P.O. Box 27, 00014, Helsinki, Finland

    Liisa Viikari

  2. ROAL, Valta-akseli, 05200, Rajamäki, Finland

    Marika Alapuranen, Terhi Puranen & Jari Vehmaanperä

  3. VTT Technical Research Centre of Finland, P.O. Box 1000, 02044, Espoo, Finland

    Matti Siika-aho

Authors
  1. Liisa Viikari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marika Alapuranen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Terhi Puranen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jari Vehmaanperä
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Matti Siika-aho
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liisa Viikari .

Editor information

Lisbeth Olsson

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Viikari, L., Alapuranen, M., Puranen, T., Vehmaanperä, J., Siika-aho, M. (2007). Thermostable Enzymes in Lignocellulose Hydrolysis . In: Olsson, L. (eds) Biofuels. Advances in Biochemical Engineering/Biotechnology, vol 108. Springer, Berlin, Heidelberg. https://doi.org/10.1007/10_2007_065

Download citation

Publish with us

Policies and ethics

Search

Navigation