World Journal of Microbiology and Biotechnology

, Volume 29, Issue 2, pp 335–345 | Cite as

Endophytes from an Australian native plant are a promising source of industrially useful enzymes

  • Bita Zaferanloo
  • Aditya Virkar
  • Peter J. Mahon
  • Enzo A. Palombo
Original Paper


Endophytes are microorganisms that live within plant tissues that are potential sources of novel bioactive compounds, including enzymes. We have identified endophytes of the Australian native plant Eremophilia longifolia which were screened for the production of industrially useful enzymes. Seventeen fungal endophytes were isolated from the leaves of E. longifolia and enzyme production was investigated within a range of pH (3.5, 5.5, 7 and 9) and temperatures (9, 25, 37 and 50 °C). Amylase was the most common enzyme encountered with numerous isolates showing production throughout the temperature and pH ranges. Protease production was also seen over the conditions tested but was more dominant at lower pH and temperature. Activity was not observed for other enzymes including ligninase, xylanase and cellobiohydrolase. Enzymes from isolates of Preussia minima, Alternaria sp. and an unclassified fungus, which showed highest activity in screening assays, were investigated further. Enzyme production was verified by zymography and the amylase activity of P. minima was found to be significantly greater than that of Aspergillus oryzae particularly in alkaline conditions and low temperature which are desirable properties for the detergent industry. This work shows that enzymes with potential use in industry can be readily identified in fungal endophytes.


Eremophilia longifolia Endophytic fungi Preussia minima Amylase Zymography 



We thank Canopus Corporation for providing the plant material used in this study.


  1. Alves MH, Campos-Takaki GM, Porto ALF, Milanez AI (2002) Screening of Mucor spp. for the production of amylase, lipase, polygalacturonase and protease. Braz J Microbiol 33:325–330Google Scholar
  2. Aly A, Debbab HA, Proksch P (2011) Fungal endophytes: unique plant inhabitants with great promises. Appl Microbiol Biotechnol 90:1829–1845CrossRefGoogle Scholar
  3. Asoodeh A, Chamani JK, Lagzian M (2010) A novel thermostable acidophilic α-amylase from a new thermophilic “Bacillus sp. Ferdowsicous” isolated from Ferdows hot mineral spring in Iran: purification and biochemical characterization. Int J Biol Macromol 46:289–297CrossRefGoogle Scholar
  4. Borges WS, Borges KB, Bonato PS, Said S, Pupo MT (2009) Endophytic fungi: natural products, enzymes and biotransformation reactions. Curr Org Chem 13:1137–1163CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Choi YW, Hodgkiss IJ, Hyde KD (2005) Enzyme production by endophytes of Bruceajavanica. J Agric Technol 1:55–66Google Scholar
  7. Ghorbel RE, Maktouf S, Massoud EB, Bejar S, Chaabouni SE (2009) New thermostable amylase from Bacillus cohnii US147 with a broad pH applicability. Appl Biochem Biotech 157:150–160CrossRefGoogle Scholar
  8. Hmidet N, Ali NEH, Haddar A, Kanoun S, Alya SK, Nasri M (2009) Alkaline proteases and thermostable α-amylase co-produced by Bacillus licheniformis NH1: characterization and potential application as detergent additive. Bio Chem Eng J 47:71–79Google Scholar
  9. Hurek BR, Hurek T (1998) Life in grasses: diazotrophicendophytes. Trends Microbiol 6:139–144CrossRefGoogle Scholar
  10. Kimura M (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120CrossRefGoogle Scholar
  11. Kirk O, Borchert TV, Fuglsan CC (2002) Industrial enzyme applications. Curr Opin Biotechnol 13:345–351CrossRefGoogle Scholar
  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  13. Laird SA, Wynberg R, Johnston S (2006) Recent trends in the biological prospecting, IP116, XXIX antarctic treaty consultative meeting (ATCM), Agenda item 18. UNU-IAS, EdinburghGoogle Scholar
  14. Lange L (2004) Tropical biodiversity, an industrial perspective. In: Bellot-Rojas M, Bernier S (eds) International expert workshop on access to genetic resources and benefit sharing. Bellot-Rojas, Cuernavaca, pp 296–300Google Scholar
  15. Márquez SS, Bills GF, Herrero N, Zabalgogeazcoa Í (2011) Non-systemic fungal endophytes of grasses. Fungal Ecol (in press)Google Scholar
  16. Matrine TF, Alarcon FJ, Lopez MD, Moyano FJ (2000) Improved detection of amylase activity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with copolymerized starch. Electrophoresis 21:2940–2943CrossRefGoogle Scholar
  17. Morgan NL, Waites MJ, Rockey JS, Higton G (2009) Industrial processes and products: microbial enzymes. In: Morgan NL, Waites MJ, Rockey JS, Higton G (eds) Industrial microbiology: an introduction. Blackwell, London, pp 133–143Google Scholar
  18. Moy M, Li HM, Sullivan R, White JF, Belanger FC (2002) Endophytic fungal β-1, 6-glucanase expression in the infected host grass. Plant Physiol 130:1298–1308CrossRefGoogle Scholar
  19. Nguyen QD, Szabo JM-R, Claeyssens M, Stals I, Hoschke A (2002) Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626. Enzym Microb Tech 31:345–352CrossRefGoogle Scholar
  20. Oses R, Valenzuela S, Freer J, Baeza J, Rodriguez J (2006) Evaluation of fungal endophytes for lignocellulolytic enzyme production and wood biodegradation. Int Biodeter Biodegr 57:129–135CrossRefGoogle Scholar
  21. Peterson RA, Bradner JR, Roberts TH, Nevalainen KMH (2009) Fungi from koala (Phascolarctoscinereus) faecesexhibit a broad range of enzyme activities against recalcitrant substrates. Lett Appl Microbiol 48:218–225CrossRefGoogle Scholar
  22. Peterson R, Grinyer J, Nevalainen H (2011) Extracellular hydrolase profiles of fungi isolated from koala faeces invite biotechnological interest. Mycol Prog 10:207–218CrossRefGoogle Scholar
  23. Schilz B, Boyle C, Draeger S, Rommert AK, Krohn K (2002) Endophytic fungi: a source of novel biologically active secondary metabolites. Mycol Res 106:996–1004CrossRefGoogle Scholar
  24. Stamford TL, Stamford NP, Coelho LC, Araujo JM (2001) Production and characterization of a thermostable α-amylase from Nocardiopsis sp. endophyte of yam bean. Bioresour Technol 77:137–141CrossRefGoogle Scholar
  25. Strobel G, Daisy B (2003) Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol R 67:491–502CrossRefGoogle Scholar
  26. Strobel G, Daisy B, Castillo U, Harper J (2004) Natural products from endophytic microorganisms. J Nat Prod 62:257–268CrossRefGoogle Scholar
  27. Tamura K, Dudley J, Nei S, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599CrossRefGoogle Scholar
  28. Tan RX, Zou WX (2001) Endophytes: a rich source of functional metabolites. Nat Prod Rep 18:448–459CrossRefGoogle Scholar
  29. Teter SA, Cherry JR (2005) Improving cellulose hydrolysis with new cellulase compositions. In: American Institute of Chemical Engineers (AIChE) Annual Meeting Conference Proceedings, Cincinnati, pp 12027–12033Google Scholar
  30. Tran HBQ, McRae JM, Lynch F, Palombo EA (2010) Identification and bioactive properties of endophytic fungi isolated from phyllodes of Acacia species. In: Méndez-Vilas A (ed) Current research, technology and education topics in applied microbiology and microbial biotechnology, no 2, vol 1. Formatex, Badajoz, pp 377–382Google Scholar
  31. Yu H, Zhang L, Li L, Zheng C, Guo L, Li W, Sun P, Qin L (2010) Recent developments and future prospects of antimicrobial metabolites produced by endophytes. Microbiol Res 165:437–449CrossRefGoogle Scholar
  32. Zhang HW, Song YC, Tan RX (2006) Biology and chemistry of endophytes. Nat Prod Rep 23:753–771CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Bita Zaferanloo
    • 1
  • Aditya Virkar
    • 1
  • Peter J. Mahon
    • 1
  • Enzo A. Palombo
    • 1
  1. 1.Faculty of Life and Social Sciences, Environment and Biotechnology CentreSwinburne University of TechnologyHawthornAustralia

Personalised recommendations