Polar Biology

, Volume 41, Issue 12, pp 2511–2521 | Cite as

Production of cold-adapted enzymes by filamentous fungi from King George Island, Antarctica

  • Alysson Wagner Fernandes Duarte
  • Mariana Blanco Barato
  • Fernando Suzigan Nobre
  • Danilo Augusto Polezel
  • Tássio Brito de Oliveira
  • Juliana Aparecida dos Santos
  • André Rodrigues
  • Lara Durães SetteEmail author
Original Paper


Antarctic environments are characterized by polar climate, making it difficult for the development of any form of life. The biogeochemical cycles and food web in such restrictive environments may be exclusively formed by microorganisms. Polar mycological studies are recent and there is much to know about the diversity and genetic resources of these microorganisms. In this sense, the molecular taxonomic approach was applied to identify 129 fungal isolates from marine and terrestrial samples collected from the King George Island (South Shetland Islands, Maritime Antarctic). Additionally, the production of cold-adapted enzymes by these microorganisms was evaluated. Among the 129 isolates, 69.0% were identified by ITS-sequencing and affiliated into 12 genera. Cadophora, Geomyces, Penicillium, Cosmospora, and Cladosporium were the most abundant genera. Representatives of Cosmospora were isolated only from terrestrial samples, while representatives of the others genera were recovered from marine and terrestrial samples. A total of 29, 19, and 74 isolates were able to produce ligninolytic enzymes, xylanase, and l-asparaginase, respectively. Representatives of Cadophora showed great ability to produce lignin peroxidase (LiP) and laccase at 15.0 °C in liquid medium, while representatives of Penicillium and non-identified fungi were the best producers of xylanase and l-asparaginase at 20.0 °C. The high number of fungi able to produce enzymes at moderate temperature reveals their potential for industrial production and biotechnological applications. The present study broadens the knowledge of fungal diversity associated with the southern polar region. Additionally, data from molecular taxonomy suggest that two filamentous fungi may be considered as potential new species.


Extremophiles Microbial biotechnology Ligninolytic enzymes l-Asparaginase Xylanase 



This study was financed by Fundação de Amparo à Pesquisa do Estado de São Paulo (Grants 2013/19486-0, 2016/07957-7). MBB and JAS thank the Coordenação de Aperfeiçoamento Pessoal de Nível Superior (CAPES) for their scholarships. LDS thanks the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the Productivity Fellowship (303145/2016-1) and the Brazilian Antarctic Program (PROANTAR). The authors thank Professor Eduardo C. M. Hajdu and Dr. Itamar S. de Melo for the samplings of marine invertebrates and ornithogenic soil, respectively.

Compliance of ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

300_2018_2387_MOESM1_ESM.pdf (2.4 mb)
Supplementary material 1 (PDF 2418 kb) Electronic Supplementary Material: Online Resource 1. Table S1 Data related to the molecular identification of filamentous fungi from King George Island: numbers of pb analyzed and GenBank accession numbers; Fig. S1 Phylogenetic analysis of the sequences of fungi from King George Island, compared with type and closest sequences of the species following BLAST analysis. (A) Acremonium; (B) Cadophora; (C) Cercospora; (D) Cladosporium; (E) Cosmospora; (F) Geomyces and Pseudogymnoascus; (G) Oidiodendron; (H) Pseudeurotium; (I) Thelebolus. The trees were constructed based on the ITS region using MEGA v. 6.0 by the neighbor-joining algorithm, Kimura 2-parameters as the nucleotide substitution model and 1000 bootstrap pseudo-replicates (values lower than 50% are not shown). Sites containing gaps were excluded from the analysis. The scale bar indicates substitutions per site. All sequences were retrieved from GenBank (accessions are shown in parentheses). CBS (Centraalbureau voor Shimmelcutures), ATCC (American Type Culture Collection), IMI (International Mycological Institute), UAMH (University of Alberta Microfungus Collection and Herbarium) and MUCL (Mycothèque de l´Université Catholique de Louvain) voucher accessions are also shown. T: type strain; NT: neotype strain.


  1. Ali SH, Alias SA, Siang HY, Smykla J, Pang KL, Guo SY, Convey P (2013) Studies on diversity of soil microfungi in the Hornsund area, Spitsbergen. Pol Polar Res 34:39–54CrossRefGoogle Scholar
  2. Arenz BE, Blanchette RA (2009) Investigations of fungal diversity in wooden structures and soils at historic sites on the Antarctic Peninsula. Can J Microbiol 55:46–56CrossRefGoogle Scholar
  3. Arenz BE, Held BW, Jurges JA, Farrell RL, Blanchette RA (2006) Fungal diversity in soils and historic wood from the Ross sea region of Antarctica. Soil Biol Biochem 38:3057–3064CrossRefGoogle Scholar
  4. Bailey MJ, Biely P, Poutanen K (1992) Interlaboratory testing of methods for assay of xylanase activity. J Biotechnol 23:257–270CrossRefGoogle Scholar
  5. Bajpai P (2004) Biological bleaching of chemical pulps. Crit Rev Biotechnol 24:1–58CrossRefGoogle Scholar
  6. Bonugli-Santos RC, Durrant LR, Sette LD (2010a) Laccase activity and putative laccase genes in marine-derived basidiomycetes. Mycol Res 114:863–872Google Scholar
  7. Bonugli-Santos RC, Durrant LR, Sette LD (2010b) Production of laccase, manganese peroxidase and lignin peroxidase by Brazilian marine-derived fungi. Enz Microb Technol 46:17–37CrossRefGoogle Scholar
  8. Bradner JR, Gillings M, Nevalainen KMH (1999) Qualitative assessment of hydrolytic activities in antarctic microfungi grown at different temperatures on solid media. World J Microbiol Biotechnol 15:131–132CrossRefGoogle Scholar
  9. Bugni TS, Ireland CM (2004) Marine-derived fungi: a chemically and biologically diverse group of microorganisms. Nat Prod Rep 21:143–163CrossRefGoogle Scholar
  10. Buswell JK, Cai YJ, Chang ST (1995) Effect of nutrient nitrogen on manganese peroxidase and laccase production by Lentinula (Lentinus) edodes. FEMS Microbiol Lett 128:81–88CrossRefGoogle Scholar
  11. Costa RR (2014) Perfil Enzimático e Potencial Biotecnológico de Fungos Isolados de Jardins de Fungo das Formigas Cortadeiras. Dissertation. Universidade Estadual Paulista “Julio de Mesquita Filho”. Rio Claro, São PauloGoogle Scholar
  12. Courtin CM, Delcour JA (2002) Arabinoxylans and endoxylanases in wheat flour bread-making. J Cereal Sci 35:225–243CrossRefGoogle Scholar
  13. Cowieson AJ, Hruby M, Pierson EE (2006) Evolving enzyme technology: impact on commercial poultry nutrition. Nutr Res Rev 19:90–103CrossRefGoogle Scholar
  14. De Menezes GC, Godinho VM, Porto BA, Gonçalves VN, Rosa LH (2017) Antarctomyces pellizariae sp. nov., a new, endemic, blue, snow resident psychrophilic ascomycete fungus from Antarctica. Extremophiles 21:259–269CrossRefGoogle Scholar
  15. Del-Cid A, Ubilla P, Rayanal MC, Medina E, Vaca I, Levicán G, Evzaquirre J, Chávez R (2014) Cold-active xylanase produced by fungi associated with Antarctic marine sponges. Appl Biochem Biotechnol 172:524–532CrossRefGoogle Scholar
  16. Dreesens LL, Lee CK, Cary SG (2014) The distribution and identity of edaphic fungi in the McMurdo Dry Valleys. Biology 3:466–483CrossRefGoogle Scholar
  17. Duarte AWF, Dayo-Owoyemi I, Nobre FS, Pagnocca FC, Chaud LSC, Pessoa JR, Felipe MGA, Sette LD (2013) Taxonomic assessment and enzymes production by yeasts isolated from marine and terrestrial Antarctic samples. Extremophiles 17:1023–1035CrossRefGoogle Scholar
  18. Duarte AWF, Passarini MR, Delforno TP, Pellizzari FM, Cipro CVZ, Montone RC, Petry MV, Putzke J, Rosa LH, Sette LD (2016) Yeasts from macroalgae and lichens that inhabit the South Shetland Islands, Antarctica. Environ Microbiol Rep 8:874–886CrossRefGoogle Scholar
  19. Duarte AWF, Santos Dos, Vianna MV, Vieira JMF, Mallagutti VH, Inforsato FJ, Wentzel LCP, Lario LD, Rodrigues A, Pagnocca FC, Pessoa JR, Sette LD (2018) Cold-adapted enzymes produced by fungi from terrestrial and marine Antarctic environments. Crit Rev Biotechnol 38:600–619CrossRefGoogle Scholar
  20. Feller G, Gerday C (2003) Psychrophilic enzymes: hot topics in cold adaptation. Nature 1:200–208Google Scholar
  21. Gerardo NM, Currie CR, Price SL, Mueller UG (2004) Exploiting a mutualism: parasite specialization on cultivars within the fungus-growing ants symbiosis. Proc R Soc Lond B Bio Sci 271:1791–1798CrossRefGoogle Scholar
  22. Gerginova MG, Peneva NM, Krumova ET, Alexieva ZA (2013) Biodegradation ability of fungal strains isolated from Antarctic towards pah. In: Proceedings of the 13th international conference of environmental science and technology Athens, Greece, 5–7 SeptGoogle Scholar
  23. Godinho VM, Furbino LE, Santiago IF, Pellizzari FM, Yokoya N, Pupo D, Alves TMA, Sales PA, Romanha AJ, Zani CL, Cantrell CL, Rosa CA, Rosa LH (2013) Diversity and bioprospecting of fungal communities associated with endemic and cold-adapted macroalgae in Antarctica. ISME J 7:1434–1451CrossRefGoogle Scholar
  24. Gonçalves VN, Vaz AB, Rosa CA, Rosa LH (2012) Diversity and distribution of fungal communities in lakes of Antarctica. FEMS Microbiol Ecol 82:459–471CrossRefGoogle Scholar
  25. Gonçalves VN, Campos LS, Melo IS, Pellizari VH, Rosa CA, Rosa LH (2013) Penicillium solitum: a mesophilic, psychrotolerant fungus present in marine sediments from Antarctica. Polar Biol 36:1823–1831CrossRefGoogle Scholar
  26. Gonçalves VN, Carvalho CR, Johann S, Mendes G, Alves TMA, Zani CL, Junior PAS, Murta SMF, Romanha AJ, Cantrell CL, Rosa CA, Rosa LH (2015) Antibacterial, antifungal and antiprotozoal activities of fungal communities present in different substrates from Antarctica. Polar Biol 38:1143–1152CrossRefGoogle Scholar
  27. Gräfenhan T, Schroers JH, Nirenberg HI, Seifert KA (2011) An overview of the taxonomy, phylogeny, and typification of nectriaceous fungi in Cosmospora, Acremonium, Fusarium, Stilbella, and Volutella. Stud Mycol 68:79–113CrossRefGoogle Scholar
  28. Granström TB, Izumori K, Leisola M (2007) A rare sugar xylitol. Part II: biotechnological production and future applications of xylitol. Appl Microbiol Biotechnol 74:273–276CrossRefGoogle Scholar
  29. Gulati R, Saxena R, Gupta RA (1997) Rapid plate assay for screening L-asparaginase producing microorganisms. Lett Appl Microbiol 24:23–26CrossRefGoogle Scholar
  30. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98Google Scholar
  31. Held BW, Jurgens JA, Duncan SM, Farrell RL, Blanchette RA (2006) Assessment of fungal diversity and deterioration in a wooden structure at New Harbor, Antarctica. Polar Biol 29:526–531CrossRefGoogle Scholar
  32. Hoog GS, Gottlich E, Platas G, Genilloud O, Leotta G, Brummelen J (2005) Evolution, taxonomy and ecology of the genus Thelebolus in Antarctica. Stud Mycol 51:33–76Google Scholar
  33. Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Bio Evol 30:772–780CrossRefGoogle Scholar
  34. Kerry E (1990) Microorganisms colonizing plants and soil subjected to different degrees of human activity, including petroleum contamination, in the Vestfoid Hills and MacRobertson Land, Antarctica. Polar Biol 10:423–430Google Scholar
  35. Laich F, Vaca I, Chávez R (2013) Rhodotorula portillonensis sp. nov., a bassidiomycetous yeast isolated from Antarctic shallow-water marine sediment. Int J Syst Evol Microbiol 63:3884–3891CrossRefGoogle Scholar
  36. Laich F, Chávez R, Vaca I (2014) Leucosporidium escuderoi f.a., sp. nov., a basidiomycetous yeast associated with an Antarctic marine sponge. Antonie Van Leeuwenhoek 105:593–601CrossRefGoogle Scholar
  37. Litova K, Gerginova M, Peneva N, Manasiev J, Alexieva Z (2014) Growth of Antarctic fungal strains on phenol at low temperatures. J BioSci Biotech 1:43–46Google Scholar
  38. Loperena L, Sonia V, Varela H, Lupo S, Bergalli A, Guigou M, Pellegrino A, Bernardo A, Calviño A, Rivas F, Batista S (2012) Extracellular enzymes produced by microorganisms isolated from maritime Antarctica. World J Microbiol Biotechnol 28:2249–2256CrossRefGoogle Scholar
  39. Lopes AM, Oliveira-Nascimento L, Ribeiro A, Tairum-Jr AC, Breyer CA, Oliveira MA, Monteiro G, Souza-Motta CM, Magalhães PO, Avendaño JGF, Cavaco-Paulo AM, Mazzola PG, Rangel-Yagui CO, Sette LD, Converti A, Pessoa A (2017) Therapeutic l-asparaginase: upstream, downstream and beyond. Crit Rev Biotechnol 37:82–99CrossRefGoogle Scholar
  40. Ludley KE, Robinson CH (2008) Decomposer’ basidiomycota in Arctic and Antarctic ecosystems. Soil Biol Biochem 40:11–29CrossRefGoogle Scholar
  41. Margesin R, Feller G, Gerday C, Russell N (2002) Cold-adapted microorganisms: adaptation strategies and biotechnological potential. In: Bitton (ed) The encyclopedia of environmental microbiology. Wiley, New York, pp 871–885Google Scholar
  42. Mcrae CF, Hocking AD, Seppelt RD (1999) Penicillium species from terrestrial habitats in the Windmill Islands, East Antarctica, including a new species, Penicillium antarcticum. Polar Biol 21:97–111CrossRefGoogle Scholar
  43. Menezes CB, Bonugli-Santos RC, Miqueletto PB, Passarini MRZ, Silva CHD, Justo MR, Leal RR, Fantinatti-Garboggini F, Oliveira VM, Berlinck RGS, Sette LD (2010) Microbial diversity associated with algae, ascidians and sponges from the north coast of São Paulo state, Brazil. Microbiol Res 165:466–482CrossRefGoogle Scholar
  44. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428CrossRefGoogle Scholar
  45. Moller EM, Bahnweg G, Sandermann H, Geiger HH (1992) A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi fruit bodies and infected plant tissues. Nucleic Acids Res 20:6115e6116CrossRefGoogle Scholar
  46. Onofri S, Selbmann L, Zucconi SP (2004) Antarctic microfungi as models for exobiology. Planet Space Sci 52:229–237CrossRefGoogle Scholar
  47. Pesciaroli C, Cupini F, Selbmann L, Barghini P, Fenice M (2012) Temperature preferences of bacteria isolated from seawater collected in Kandalaksha Bay, White Sea, Russia. Polar Biol 35:435–445CrossRefGoogle Scholar
  48. Polizeli ML, Rizzatti AC, Monti R, Terenzi HF, Jorge JA, Amorim DS (2005) Xylanases from fungi: properties and industrial applications. Appl Microbiol Biotechnol 67:577–591CrossRefGoogle Scholar
  49. Pudasaini S, Wilson J, Ji M, van Dorst J, Snape I, Palmer AS, Burns BP, Ferrari (2017) Microbial diversity of browning peninsula, Eastern Antarctica revealed using molecular and cultivation methods. Front Microbiol 8:591CrossRefGoogle Scholar
  50. Ruisi S, Barreca D, Selbmann L, Zucconi L, Onofri S (2007) Fungi in Antarctica. Rev Environ Sci Biotechnol 6:127–141CrossRefGoogle Scholar
  51. Santiago IF, Soares MA, Rosa CA, Rosa LA (2016) Lichensphere: a protected natural microhabitat of the non-lichenised fungal communities living in extreme environments of Antarctica. Extremophiles 19:1087–1097CrossRefGoogle Scholar
  52. Siddiqui KS, Cavicchioli R (2006) Cold-adpated enzymes. Annu Rev Biochem 75:403–433CrossRefGoogle Scholar
  53. Sonjak S, Frisvad JC, Gunde-Cimerman N (2006) Penicillium mycobiota in Arctic subglacial ice. Microbial Ecol 52:207–216CrossRefGoogle Scholar
  54. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefGoogle Scholar
  55. Theantana T, Hyde KD, Lumyong S (2007) Asparaginase production by Endophytic fungi isolated from some Thai medicinal plants. KMITL Sci Tech 7:13-1Google Scholar
  56. Tien M, Kirk TK (1984) Lignin-degrading enzyme from Phanerochaete chrysosporium: purification, characterization and catalytic properties of unique H2O2 requiring oxygenase. Proc Natl Aca Sci USA 81:2280–2284CrossRefGoogle Scholar
  57. Tindall BJ (2004) Prokaryotic diversity in the Antarctic: the tip of the iceberg. Microb Ecol 47:271–283CrossRefGoogle Scholar
  58. Turchetti B, Selbmann L, Blanchette RA, Di Mauro S, Marchegiani E, Zucconi L, Arenz BE, Buzzini P (2015) Cryptococcus vaughanmartiniae sp. nov. and Cryptococcus onofrii sp. nov.: two new species isolated from worldwide cold environments. Extremophiles 19:149–159CrossRefGoogle Scholar
  59. Verma AK, Ragggukkumar C, Verma P, Shouche YS, Nail CG (2010) Four marine-derived fungi for bioremediation of raw textile mill effluents. Biodegradation 21:217–233CrossRefGoogle Scholar
  60. Vincent WF (2000) Evolutionary origins of Antarctic microbiota: invasion, selection and endemism. Antarct Sci 12:374–385CrossRefGoogle Scholar
  61. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Shinsky TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, New York, pp 315–322Google Scholar
  62. Zhang T, Zhang YQ, Liu HY, Zhao LX, Yu LY (2014) Cryptococcus fildesensis sp. nov., a psycrophilic basidiomycetous yeast isolated from Antarctic moss. Int J Syst Evol Microbiol 64:675–679CrossRefGoogle Scholar
  63. Zhdanova NN, Zakaarchenko VA, Vember VV, Nakonechnaya LT (2000) Fungi from Chernobyl: mycobiota of the inner regions of the containment structures of the damaged nuclear reactor. Mycol Res 104:1421–1426CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Alysson Wagner Fernandes Duarte
    • 1
  • Mariana Blanco Barato
    • 2
  • Fernando Suzigan Nobre
    • 2
  • Danilo Augusto Polezel
    • 3
  • Tássio Brito de Oliveira
    • 3
  • Juliana Aparecida dos Santos
    • 3
  • André Rodrigues
    • 3
  • Lara Durães Sette
    • 2
    • 3
    Email author
  1. 1.Federal University of AlagoasArapiracaBrazil
  2. 2.Division of Microbial ResourcesChemical, Biological and Agricultural Pluridisciplinary Research Center (CPQBA)/Campinas State University (UNICAMP)PaulíniaBrazil
  3. 3.Department of Biochemistry and Microbiology, Institute of BiosciencesSão Paulo State University (UNESP)Rio ClaroBrazil

Personalised recommendations