Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

From Agricultural Waste to Biofuel: Enzymatic Potential of a Bacterial Isolate Streptomyces fulvissimus CKS7 for Bioethanol Production

  • 29 Accesses



To avoid a negative environmental and economic impact of agricultural wastes, and following the principles of circular economy, the reuse of agricultural wastes is necessary. For this purpose, isolation of novel microorganisms with potential biotechnological application is recommended. The current researches in bioethanol production are aimed to reduce the production costs using low-cost substrates and in-house produced enzymes by novel isolated microorganisms. In line with this, in this study valorization of these agricultural by-products by novel isolate S. fulvissimus CKS7 to biotechnological value added products was done.


Standard microbiological methods were used for the isolation and characterization of strain. Enzymes activities were determinated using DNS method while, the ethanol concentration was determined based on the density of the alcohol distillate at 20 °C.


The maximal enzymatic activities for amylase, cellulases (carboxymethyl cellulase and Avicelase), pectinase and xylanase were achieved using rye bran as a waste substrate for CKS7 growth. Obtained crude bacterial enzymes were used for enzymatic hydrolysis of lignocellulosic materials including horsetail waste, yellow gentian waste, corn stover, cotton material and corona pre-treated cotton material. The maximum yield of reducing sugars was obtained on horsetail waste and corona pre-treated cotton material. Waste brewer’s yeast Saccharomyces cerevisiae was successfully used for the production of bioethanol using horsetail waste hydrolysate and corona pre-treated cotton material hydrolysate.


The obtained results showed that bacterial strain CKS7 has a significant, still unexplored enzymatic potential that could be used to achieve a cleaner, environmental friendly and economically acceptable biofuel production.

Graphic Abstract

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

Fig. 1
Fig. 2


  1. 1.

    Fischer, C.N.: Exploring the diversity of the microbial world. Yale J. Biol. Med. 84(1), 55–58 (2011)

  2. 2.

    Alves, P.D.D., de Faria Siqueira, F., Facchin, S., Horta, C.C.R., Victória, J.M.N., Kalapothakis, E.: Survey of microbial enzymes in soil, water, and plant microenvironments. Open Microbiol. J. 8, 25–31 (2014)

  3. 3.

    Tsegaye, B., Balomajumder, C., Roy, P.: Isolation and characterization of novel lignolytic, cellulolytic, and hemicellulolytic bacteria from wood-feeding Termite Cryptotermes brevis. Int. Microbiol. 22, 29–39 (2019)

  4. 4.

    Méndez-Vilas, A.: Communicating Current Research and Educational Topics and Trends in Applied Microbiology, vol. 2. Formatex, Badajoz (2007)

  5. 5.

    Tsegaye, B., Balomajumder, C., Roy, P.: Biodelignification and hydrolysis of rice straw by novel bacteria isolated from wood feeding termite. 3 Biotech 8, 447–458 (2018)

  6. 6.

    Fendrihan, S.: Bioproducts with living microorganisms used in agriculture. Res. J. Pharm. Phytochem. 9, 10–14 (2016)

  7. 7.

    Skujiņš, J., Burns, R.: Extracellular enzymes in soil. CRC Crit. Rev. Microbiol. 4, 383–421 (1976)

  8. 8.

    Ravindran, R., Hassan, S., Williams, G., Jaiswal, A.: A review on bioconversion of agro-industrial wastes to industrially important enzymes. BIOEBG 5, 93–113 (2018)

  9. 9.

    Demir, H., Tari, C.: Bioconversion of wheat bran for polygalacturonase production by Aspergillus sojae in tray type solid-state fermentation. Int. Biodeter. Biodegr. 106, 60–66 (2016)

  10. 10.

    Jecu, L.: Solid state fermentation of agricultural wastes for endoglucanase production. Ind. Crop. Prod. 11, 1–5 (2000)

  11. 11.

    Pothiraj, C., Kanmani, P., Balaji, P.: Bioconversion of lignocellulose materials. Mycobiology 34, 159–165 (2006)

  12. 12.

    Pandey, A., Selvakumar, P., Soccol, C.R., Nigam, P.: Solid state fermentation for the production of industrial enzymes. Curr. Sci. 77, 149–162 (1999)

  13. 13.

    Auer, L., Lazuka, A., Sillam-Dussès, D., Miambi, E., O'Donohue, M., Hernandez-Raquet, G.: Uncovering the potential of termite gut microbiome for lignocellulose bioconversion in anaerobic batch bioreactors. Front. Microbiol. 8, 1637–2623 (2017)

  14. 14.

    Brune, A.: Symbiotic digestion of lignocellulose in termite guts. Nat. Rev. Microbiol. 12(3), 168–180 (2014)

  15. 15.

    Razmovski, R., Vučurović, V.: Bioethanol production from sugar beet molasses and thick juice using Saccharomyces cerevisiae immobilized on maize stem ground tissue. Fuel 92, 1–8 (2012)

  16. 16.

    Domínguez-Bocanegra, A.R., Torres-Muñoz, J.A., López, R.A.: Production of bioethanol from agro-industrial wastes. Fuel 149, 85–89 (2015)

  17. 17.

    Johnson, E.: Integrated enzyme production lowers the cost of cellulosic ethanol. Biofuel. Bioprod. Bior. 10, 164–174 (2016)

  18. 18.

    El-Naggar, N., Deraz, S., Khalil, A.: Bioethanol production from lignocellulosic feedstocks based on enzymatic hydrolysis: current status and recent developments. Biotechnology 13, 1–21 (2014)

  19. 19.

    de Lima Procópio, R.E., da Silva, I.R., Martins, M.K., de Azevedo, J.L., de Araújo, J.M.: Antibiotics produced by Streptomyces. Braz. J. Infect. Dis. 16, 466–471 (2012)

  20. 20.

    Sathya, R., Ushadevi, T.: Industrially important enzymes producing streptomyces species from mangrove sediments. Int. J. Pharm. Pharm. Sci. 6, 233–237 (2014)

  21. 21.

    Sinjaroonsak, S., Chaiyaso, T., Aran, H.: Optimization of cellulase and xylanase productions by Streptomyces thermocoprophilus TC13W using low cost pretreated oil palm empty fruit bunch. Waste Biomass Valori. (2019). https://doi.org/10.1007/s12649-019-00720-y

  22. 22.

    Buntić, A., Pavlović, M., Šiler-Marinković, S., Dimitrijević-Branković, S.: Biological treatment of colored wastewater by Streptomyces fulvissimus CKS 7. Water Sci. Technol. 73, 2231–2236 (2016)

  23. 23.

    Marković, D., Deeks, C., Nunney, T., Radovanović, Ž., Radoičić, M., Šaponjić, Z., Radetić, M.: Antibacterial activity of Cu-based nanoparticles synthesized on the cotton fabrics modified with polycarboxylic acids. Carbohydr. Polym. 200, 173–182 (2018)

  24. 24.

    Mihajlovski, K.R., Carević, M.B., Dević, M.L., Šiler-Marinković, S., Rajilić-Stojanović, M.D., Dimitrijević-Branković, S.: Lignocellulosic waste material as substrate for Avicelase production by a new strain of Paenibacillus chitinolyticus CKS1. Int. Biodeter. 104, 426–434 (2015)

  25. 25.

    Kasana, R.C., Salwan, R., Dhar, H., Dutt, S., Gulati, A.: A rapid and easy method for the detection of microbial cellulases on agar plates using Gram’s iodine. Curr. Microbiol. 57(5), 503–507 (2008)

  26. 26.

    Miller, G.L.: Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31, 426–428 (1959)

  27. 27.

    Mihajlovski, K., Radovanović, Ž., Carević, M., Dimitrijević-Branković, S.: Valorization of damaged rice grains: Valorization of damaged rice grains: Optimization of bioethanol production by waste brewer’s yeast using an amylolytic potential from the Paenibacillus chitinolyticus CKS1. Fuel 224, 591–599 (2018)

  28. 28.

    Horwitz, W., Chichilo, P., Reynolds, H.: Official Methods of Analysis of the Association of Official Analytical Chemists. Association of Official Analytical Chemists, Washington (1970)

  29. 29.

    Sanjivkumar, M., Brindhashini, A., Deivakumari, M., Palavesam, A., Immanuel, G.: Investigation on saccharification and bioethanol production from pretreated agro-residues using a mangrove associated actinobacterium Streptomyces variabilis (MAB3). Waste Biomass Valori. 9, 969–984 (2018)

  30. 30.

    Ikeda, H., Ishikawa, J., Hanamoto, A., Shinose, M., Kikuchi, H., Shiba, T., Sakaki, Y., Hattori, M., Ōmura, S.: Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat. Biotechnol. 21, 526–531 (2003)

  31. 31.

    Lynd, L.R., Weimer, P.J., Van Zyl, W.H., Pretorius, I.S.: Microbial cellulose utilization: fundamentals and biotechnology. Microbiol. Mol. Biol. Rev. 66, 506–577 (2002)

  32. 32.

    Lizardi-Jiménez, M., Hernández-Martínez, R.: Solid state fermentation (SSF): diversity of applications to valorize waste and biomass. 3 Biotech 7, 44–53 (2017)

  33. 33.

    Rodríguez Couto, S.: Exploitation of biological wastes for the production of value-added products under solid-state fermentation conditions. Biotechnol. J. 3, 859–870 (2008)

  34. 34.

    Azzeddine, B., Abdelaziz, M., Estelle, C., Mouloud, K., Nawel, B., Nabila, B., Francis, D., Said, B.: Optimization and partial characterization of endoglucanase produced by Streptomyces sp. B-PNG23. Arch. Biol. Sci. 65, 549–558 (2013)

  35. 35.

    de Lima, A.L.G., do Nascimento, R.P., da Silva Bon, E.P., Coelho, R.R.R.: Streptomyces drozdowiczii cellulase production using agro-industrial by-products and its potential use in the detergent and textile industries. Enzyme Microb. Technol. 37, 272–277 (2005)

  36. 36.

    Budihal, S., Agsar, D.: Exploration of agrowastes for the production of cellulase by Streptomyces DSK29 under submerged and solid state systems. Int. J. Curr. Microbiol. Appl. Sci 4, 681–689 (2015)

  37. 37.

    Izydorczyk, M.: Arabinoxylans. In: Philips, G.O., Williams, P.A. (eds.) Handbook of Hydrocoloids, pp. 653–692. Elsevier, New Delhi (2009)

  38. 38.

    Banaszkiewicz, T.: Nutritional value of soybean meal. In: El-Shemy, H. (ed.) Soybean and Nutrition, pp. 1–23. InTech, Rijeka (2011)

  39. 39.

    Sredanović, S., Lević, J., Đuragić, O.: Enzyme enhancement of the nutritional value of sunflower meal. Enzyme Microb. Technol. 21, 197–202 (2005)

  40. 40.

    Malathi, V., Devegowda, G.: In vitro evaluation of nonstarch polysaccharide digestibility of feed ingredients by enzymes. Poult. Sci. 80, 302–305 (2001)

  41. 41.

    Patil, N.S., Jadhav, J.P.: Enzymatic production of N-acetyl-D-glucosamine by solid state fermentation of chitinase by Penicillium ochrochloron MTCC 517 using agricultural residues. Int. Biodeter. Biodegr. 91, 9–17 (2014)

  42. 42.

    de Queiroz Brito-Cunha, C.C., de Campos, I.T.N., de Faria, F.P., Bataus, L.A.M.: Screening and xylanase production by Streptomyces sp. grown on lignocellulosic wastes. Appl. Biochem. Biotechnol. 170, 598–608 (2013)

  43. 43.

    Zheng, Y.-X., Wang, Y.-L., Pan, J., Zhang, J.-R., Dai, Y., Chen, K.-Y.: Semi-continuous production of high-activity pectinases by immobilized Rhizopus oryzae using tobacco wastewater as substrate and their utilization in the hydrolysis of pectin-containing lignocellulosic biomass at high solid content. Bioresour. Technol. 241, 1138–1144 (2017)

  44. 44.

    Mihailović, V., Mišić, D., Matić, S., Mihailović, M., Stanić, S., Vrvić, M.M., Katanić, J., Mladenović, M., Stanković, N., Boroja, T.: Comparative phytochemical analysis of Gentiana cruciata L. roots and aerial parts, and their biological activities. Ind. Crop. Prod. 73, 49–62 (2015)

  45. 45.

    Milutinović, M., Radovanović, N., Rajilić-Stojanović, M., Šiler-Marinković, S., Dimitrijević, S., Dimitrijević-Branković, S.: Microwave-assisted extraction for the recovery of antioxidants from waste Equisetum arvense. Ind. Crop. Prod. 61, 388–397 (2014)

  46. 46.

    Ximenes, E., Kim, Y., Mosier, N., Dien, B., Ladisch, M.: Inhibition of cellulases by phenols. Enzyme Microb. Technol. 46, 170–176 (2010)

  47. 47.

    Gupta, A., Verma, J.P.: Sustainable bio-ethanol production from agro-residues: a review. Renew. Sust. Energ. Rev. 41, 550–567 (2015)

  48. 48.

    Nikolić, S., Lazić, V., Veljović, Đ., Mojović, L.: Production of bioethanol from pre-treated cotton fabrics and waste cotton materials. Carbohydr. Polym. 164, 136–144 (2017)

  49. 49.

    Sun, S., Sun, S., Cao, X., Sun, R.: The role of pretreatment in improving the enzymatic hydrolysis of lignocellulosic materials. Bioresour. Technol. 199, 49–58 (2016)

  50. 50.

    Lu, X., Zhang, Y., Yang, J., Liang, Y.: Enzymatic hydrolysis of corn stover after pretreatment with dilute sulfuric acid. Chem. Eng. Technol. 30, 938–944 (2007)

  51. 51.

    Satarn, J., Lamamorphanth, W., Kamwilaisak, K.: Acid hydrolysis from corn stover for reducing sugar. Adv. Mater. Resour. 931, 1608–1613 (2014)

  52. 52.

    Panagiotou, G., Kekos, D., Macris, B.J., Christakopoulos, P.: Production of cellulolytic and xylanolytic enzymes by Fusarium oxysporum grown on corn stover in solid state fermentation. Ind. Crop. Prod. 18, 37–45 (2003)

  53. 53.

    Singhania, R.R., Agarwal, R.A., Kumar, R.P., Sukumaran, R.K.: Waste to Wealth. Springer, Singapore (2018)

  54. 54.

    Umamaheswari, M., Jayakumari, M., Maheswari, K., Subashree, M., Mala, P., Sevanthi, T., Manikandan, T.: Bioethanol production from cellulosic materials. Int. J. Curr. Res. 1, 005–011 (2010)

  55. 55.

    Idrees, M., Adnan, A., Bokhari, S.A., Qureshi, F.A.: Production of fermentable sugars by combined chemo-enzymatic hydrolysis of cellulosic material for bioethanol production. Braz. J. Chem. Eng. 31, 355–363 (2014)

Download references


The financial support for this investigation given by the Ministry of Education, Science and Technological Development of the Republic of Serbia under the project TR 31035 is gratefully acknowledged. The authors would like to thank the Dr Darka Marković for providing cotton and corona pre-treated cotton material. Also, the authors would like to thank agricultural cooperative “Mrkšićevi salaši” for obtaining corn waste.

Author information

Correspondence to Katarina Mihajlovski.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain studies with human participants or animals.

Informed Consent

Informed consent was obtained from all the individual participants included in the current study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 571 kb)

Supplementary file2 (DOCX 12 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Mihajlovski, K., Buntić, A., Milić, M. et al. From Agricultural Waste to Biofuel: Enzymatic Potential of a Bacterial Isolate Streptomyces fulvissimus CKS7 for Bioethanol Production. Waste Biomass Valor (2020). https://doi.org/10.1007/s12649-020-00960-3

Download citation


  • Solid state fermentation
  • Streptomyces fulvissimus CKS7
  • Hydrolytic enzymes production
  • Lignocellulosic waste hydrolysis
  • Bioethanol