Skip to main content
Log in

Penicillium subrubescens, a new species efficiently producing inulinase

  • Original Paper
  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

Inulin is a reserve carbohydrate in about 15 % of the flowering plants and is accumulated in underground tubers of e.g. chicory, dahlia and Jerusalem artichoke. This carbohydrate consists of linear chains of β-(2,1)-linked fructose attached to a sucrose molecule. Inulinases hydrolyse inulin into fructose and glucose. To find efficient inulin degrading fungi, 126 fungal strains from the Fungal Biotechnology Culture Collection (FBCC) at University of Helsinki and 74 freshly isolated strains from soil around Jerusalem artichoke tubers were screened in liquid cultures with inulin as a sole source of carbon or ground Jerusalem artichoke tubers, which contains up to 19 % (fresh weight) inulin. Inulinase and invertase activities were assayed by the dinitrosalicylic acid (DNS) method and a freshly isolated Penicillium strain originating from agricultural soil (FBCC 1632) was the most efficient inulinase producer. When it was cultivated at pH 6 and 28 °C in 2 litre bioreactors using inulin and Jerusalem artichoke as a carbon source, inulinase and invertase activities were on day 4 7.7 and 3.1 U mL−1, respectively. The released sugars analysed by TLC and HPLC showed that considerable amounts of fructose were released while the levels of oligofructans were low, indicating an exoinulinase type of activity. Taxonomic study of the inulinase producing strain showed that this isolate represents a new species belonging in Penicillium section Lanata-divaricata. This new species produces a unique combination of extrolites and is phenotypically and phylogenetically closely related to Penicillium pulvillorum. We propose the name Penicillium subrubescens sp. nov. (CBS 132785T = FBCC 1632T) for this new species.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abd Allah AbdAl-Aziz S, El-Metwally MM, Saber WEIA (2012) Molecular identification of a novel inulinolytic fungus isolated from and grown on tubers of Helianthus tuberosus and statistical screening of medium components. World J Microbiol Biotechnol 28:3245–3254

    Article  PubMed  Google Scholar 

  • Azhari R, Szlak AM, Ilan E (1989) Purification and characterization of endo- and exo-inulinase. Biotechnol Appl Biochem 11:107–117

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Christensen M (1981) Species diversity and dominance in fungal communities. In: Wicklow DT, Carroll GC (eds) The fungal community: its organization and role in the ecosystem. Marcel Dekker, New York, pp 201–232

    Google Scholar 

  • Christensen M, Frisvad JC, Tuthill DE (2000) Penicillium species diversity in soil and some taxonomic and ecological notes. In: Samson RA, Pitt JI (eds) Integration of modern taxonomic methods for Penicillium and Aspergillus classification. Harwood Academic Publishers, Amsterdam, pp 309–320

    Google Scholar 

  • Christov LP, Prior BA (1994) Enzymatic prebleaching of sulphite pulps. Appl Microbiol Biotechnol 42:492–498

    Article  CAS  Google Scholar 

  • Durieux A, Fougnies C, Jacobs H, Simon JP (2001) Metabolism of chicory fructooligosaccharides by bifidobacteria. Biotechnol Lett 23:1523–1527

    Article  CAS  Google Scholar 

  • Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797

    Article  PubMed  CAS  Google Scholar 

  • Ettalibi M, Baratti JC (1987) Purification, properties and comparison of invertase, exinulinases and endoinulinases of Aspergillus ficuum. Appl Microbiol Biotechnol 26:13–20

    Article  CAS  Google Scholar 

  • Franck A, de Leenheer L (2002) Inulin. In: Steinbüchel A, De Baets S, Vandamme E (eds) Biopolymers, vol 6., Polysaccharides II: polysaccharides from eukaryotesWiley–VCH, Weinheim, pp 439–480

    Google Scholar 

  • Frisvad JC, Thrane U (1987) Standardized high performance liquid chromatography of 182 mycotoxins and other fungal metabolites based on alkylphenone retention indices and UV–VIS spectra (diode array detection). J Chromatogr A 404:195–214

    Article  CAS  Google Scholar 

  • Houbraken J, Samson RA (2011) Phylogeny of Penicillium and the segregation of Trichocomaceae into three families. Stud Mycol 70:1–51

    Article  PubMed  CAS  Google Scholar 

  • Houbraken J, Frisvad JC, Samson RA (2011a) Taxonomy of Penicillium section Citrina. Stud Mycol 70:53–138

    Article  PubMed  CAS  Google Scholar 

  • Houbraken J, López-Quintero CA, Frisvad JC, Boekhout T, Theelen B, Esperanza Franco-Molano A, Samson RA (2011b) Penicillium araracuarense sp. nov., Penicillium elleniae sp. nov., Penicillium penarojense sp. nov., Penicillium vanderhammenii sp. nov. and Penicillium wotroi sp. nov., isolated from leaf litter. Int J Syst Evol Microbiol 61:1462–1475

    Article  PubMed  Google Scholar 

  • Houbraken J, Spierenburg H, Frisvad JC (2012) Rasamsonia, a new genus comprising thermotolerant and thermophilic Talaromyces and Geosmithia species. Antonie Van Leeuwenhoek 101:403–421

    Article  PubMed  CAS  Google Scholar 

  • Kantelinen A, Hatakka A, Viikari L (1989) Production of lignin peroxidase and laccase by Phlebia radiata. Appl Microbiol Biotechnol 31:234–239

    Article  CAS  Google Scholar 

  • Krogh KBR, Mørkeberg A, Jørgensen H, Frisvad JC, Olsson L (2004) Screening in genus Penicillium for producers of cellulolytic and xylanolytic enzymes. Appl Biochem Biotechnol 113–116:389–401

    Article  PubMed  Google Scholar 

  • Kushi RT, Monti R, Contiero J (2000) Production, purification and characterization of an extracellular inulinase from Kluyveromyces marxianus var. bulgaricus. J Ind Microbiol Biotechnol 25:63–69

    Article  CAS  Google Scholar 

  • Lafuente-Castaneda C, Martinez G, Contreras-Esquivel JC, Rodrigues-Herrera R, Prado-Barragan A, Aguilar CN (2012) Inulinase production by Penicillium citrinum ESS in submerge and solid-state cultures. Am J Agric Biol Sci 7:129–134

    Article  CAS  Google Scholar 

  • Lund F (1995) Differentiating Penicillium species by detection of indole metabolites using a filter paper method. Lett Appl Microbiol 20:228–231

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Mohammed S, Wesam IA, El-Ahmady S, El-Ahmadi N (2011) Production strategy of inulinase by Penicillium citrinum AR-IN2 on some agricultural by-products. Microbiol J 1:79–88

    Article  Google Scholar 

  • Nakamura T, Hoashi S, Nakatsu S (1978) Culture conditions for inulinase production by Aspergillus. Nippon Nogeikagaku Kaishi 52:105–110

    Article  CAS  Google Scholar 

  • Nakamura T, Ogata Y, Hamada S, Ohta K (1996) Ethanol production from Jerusalem artichoke tubers by Aspergillus niger and Saccharomyces cervisiae. J Ferment Bioeng 81:564–566

    Article  CAS  Google Scholar 

  • Nakamura T, Shitara A, Matsuda S, Matsuo T, Suiko M, Ohta K (1997) Production, purification and properties of an endoinulinase of Penicillium sp. TN-88 that liberates inulotriose. J Ferment Bioeng 84:313–318

    Article  CAS  Google Scholar 

  • Pitt JI (1980) The genus Penicillium and its teleomorphic states Eupenicillium and Talaromyces. Academic Press, London

    Google Scholar 

  • Ricca E, Calabro V, Curcio S, Lorio G (2007) The state of the art in the production of fructose from inulin enzymatic hydrolysis. Crit Rev Biotechnol 27:129–145

    Article  PubMed  CAS  Google Scholar 

  • Samson RA, Houbraken J, Thrane U, Frisvad JC, Andersen B (2010) Food and Indoor Fungi., CBS laboratory manual series 2CBS-Fungal Biodiversity Centre, Utrecht

    Google Scholar 

  • Scarborough AM (1970) The Soil Microfungi of a Colorado Grassland. MS Thesis, Botany Department, University of Wyoming, Laramie

  • Sharma AD, Gill PK, Bhullar SS, Singh P (2005) Improvement in inulinase production by simultaneous action of physical and chemical mutagenesis in Penicillium purpurogenum. World J Microbiol Biotechnol 21:929–932

    Article  CAS  Google Scholar 

  • Sharma AD, Kainth S, Gill PK (2006) Inulinase production using garlic (Allium sativum) powder as potential substrate in Streptomyces sp. J Food Eng 77:486–491

    Article  CAS  Google Scholar 

  • Sonjak S, Uršič V, Frisvad JC, Gunde-Cimerman N (2007) Penicillium svalbardense, a new species from Arctic glacial ice. Antonie Van Leeuwenhoek 92:43–51

    Article  PubMed  CAS  Google Scholar 

  • Stamatakis A, Hoover P, Rougemont J (2008) A rapid bootstrap algorithm for the RAxML web-servers. Syst Biol 57:758–771

    Article  PubMed  Google Scholar 

  • 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 28:2731–2739

    Article  PubMed  CAS  Google Scholar 

  • Tuthill DE, Frisvad JC, Christensen M (2001) Systematics of Penicillium simplicissimum based on rDNA sequences, morphology and secondary metabolites. Mycologia 93:298–308

    Article  CAS  Google Scholar 

  • Uzunova K, Vassileva A, Ivanova V, Spasova D, Tonkova A (2002) Thermostable exo-inulinase production by semicontinuous cultivation of membrane-immobilized Bacillus sp. 11 cells. Process Biochem 37:863–868

    Article  CAS  Google Scholar 

  • Vandamme EJ, Derycke DG (1983) Microbial inulinases: fermentation process, properties and application. Adv Appl Microbiol 29:139–176

    Article  PubMed  CAS  Google Scholar 

  • Wang L, Liu Z, Zhao G, Long X, Chong L (2007) Isolation of inulinase production strain and optimization of its fermentation condition. J Nanjing Agric Univ 30:73–77

    CAS  Google Scholar 

Download references

Acknowledgments

The work was funded by CIMO Grant (TM-11-7576) and EMbaRC open access grant. Hilkka Koponen is acknowledged for the preliminary identification of the Penicillium strains and Ronald de Vries for his valuable suggestions. Ellen Kirstine Lyhne and Martin Meijer are thanked for their technical contributions and JCF thanks the Danish Council for Strategic Research to the project MycoFuelChem.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J. Houbraken.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 134 kb)

Supplementary material 2 (PDF 97 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mansouri, S., Houbraken, J., Samson, R.A. et al. Penicillium subrubescens, a new species efficiently producing inulinase. Antonie van Leeuwenhoek 103, 1343–1357 (2013). https://doi.org/10.1007/s10482-013-9915-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10482-013-9915-3

Keywords

Navigation