Abstract
The morphological type of a microorganism generally influences its metabolite production. In the present study, we investigated the effects of the mycelial morphology of shiitake (Lentinula edodes) on the production of 2-mercaptohistidine trimethylbetaine (ergothioneine, ESH) during liquid fermentation. Analyses of the distribution of ESH in mycelial cells of different morphological types revealed that the ESH content of pellets obtained from the liquid fermentation media was much greater than the content in the free mycelia and clumps. The concentration of ESH in pellets on day 15 of liquid fermentation reached 0.79 mg/g dry weight (DW), which is approximately three times the concentration found in mycelia clumps (0.28 mg/g DW) and free mycelia (0.31 mg/g DW). Macroscopic image analysis of the development and morphological changes of the pellets during a liquid fermentation period of up to 25 days indicated that pellet growth showed a highly positive correlation with the increase in ESH concentration (r 2 = 0.9851). A reduced agitation rate of 50 rpm for the culture medium was suitable for pellet formation and size enlargement. The results obtained in this work would be helpful in guiding the intentional manipulation of the distribution and enrichment of ESH in L. edodes through changes in liquid fermentation conditions.
Similar content being viewed by others
References
Alvim FC, Mattos EM, Pirovani CP, Gramacho K, Pungartnik C, Brendel M, Cascardo JCM, Vincentz M (2009) Carbon source-induced changes in the physiology of the cacao pathogen Moniliophthora perniciosa (Basidiomycetes) affect mycelial morphology and secretion of necrosis-inducing proteins. Genet Mol Res 8:1035–1050
Baets SD, Laing SD, Francois C, Vandamme E (2002) Optimization of exopolysaccharide production by Tremella mesenterica NRRL Y-6158 through implementation of fed-batch fermentation. J Ind Microbiol Biotechnol 29:181–184
Berovic M, Cimerman A (1982) Redox potential in submerged citric acid fermentation. Appl Microbiol Biotechnol 16:185–188
Berovic M, Cimerman A, Steiner W, Koloni T (1991) Submerged citric acid fermentation: rheological properties of Aspersillus niger broth in a stirred tank reactor. Appl Microbiol Biotechnol 34:579–581
Braun S, Vecht-Lifshitz SE (1991) Mycelial morphology and metabolite production. Trends Biotechnol 9:63–68
Casas López JL, Pérez Sánchez JA, Sevilla Fernández JM, Rodríguez Porcel EM, Chisti YJ (2005) Pellet morphology, culture rheology and lovastatin production in cultures of Aspergillus terreus. J Biotechnol 116:61–77
Cui YQ, Okkerse WJ, van der Lans RGJM, Luyben KChAM (1998) Modeling and measurements of fungal growth and morphology in submerged fermentations. Biotechnol Bioeng 60:216–229
Dubost NJ, Beelman R, Royse D (2007) Influence of selected cultural factors and postharvest storage on ergothioneine content of common button mushroom Agaricus bisporus (J. Lge) Imbach (Agaricomycetideae). Int J Medicinal Mushrooms 9:163–176
Estrada A, Lee HJ, Beelman R, Jimenez-Gasco M, Royse D (2009) Enhancement of the antioxidants ergothioneine and selenium in Pleurotus eryngii var. eryngii basidiomata through cultural practices. World J Microbiol Biotechnol 25:1597–1607
Fazenda ML, Harvey LM, McNeil B (2010) Effects of dissolved oxygen on fungal morphology and process rheology during fed-batch processing of Ganoderma lucidum. J Microbiol Biotechnol 20:844–851
Gehrig I, Bart HJ, Anke T, Germerdonk R (1998) Influence of morphology and rheology on the production characteristics of the basidiomycete Cyathus striatus. Biotechnol Bioeng 59:525–533
Gibbs PA, Seviour RJ (1996) Does the agitation rate and/or oxygen saturation influence exopolysaccharide production by Aureobasidium pullulans in batch culture? Appl Microbiol Biotechnol 46:503–510
Gibbs PA, Seviour RJ, Schmid F (2000) Growth of filamentous fungi in submerged culture: problems and possible solutions. Crit Rev Biotechnol 20:17–48
Hartman PE (1990) Ergothioneine as antioxidant. Methods Enzymol 186:310–318
Hsieh C, Liu CJ, Tseng MH, Lo CT, Yang YC (2006) Effect of olive oil on the production of mycelial biomass and polysaccharides of Grifola frondosa under high oxygen concentration aeration. Enzyme Microb Technol 39:434–439
Hwang HJ, Kim SW, Xu CP, Choi JW, Yun JW (2004) Morphological and rheological properties of the three different species of basidiomycetes Phellinus in submerged cultures. J Appl Microbiol 96:1296–1305
Lee WY, Park EJ, Ahn JK (2009) Supplementation of methionine enhanced the ergothioneine accumulation in the Ganoderma neo-japonicum mycelia. Appl Biochem Biotechnol 158:213–221
Mattila P, Suonpää K, Piironen V (2002) Functional properties of edible mushrooms. Nutrition 16:694–696
Melville DB, Horner WH, Otken CC, Ludwig ML (1955) Studies on the origin of ergothioneine in animals. J Biol Chem 213:61–68
Metz B, Kossen NWF (1977) Biotechnology review: the growth of molds in the form of pellets. A literature review. Biotechnol Bioeng 19:781–799
Moore D, Robson GD, Trinci APJ (2011) Whole organism biotechnology. In: Moore D, Robson GD, Trinci APJ (eds) 21st century guidebook to fungi. Cambridge University Press, London, pp 451–510
Nielsen J, Johansen CL, Jacobsen M, Krabben P, Villadsen J (1995) Pellet formation and fragmentation in submerged cultures of Penicillium chrysogenum and its relation to penicillin production. Biotechnol Prog 11:93–98
Packer HL, Thomas CR (1990) Morphological measurements on filamentous microorganisms by fully automatic image analysis. Biotechnol Bioeng 35:870–881
Papagianni M (2004) Fungal morphology and metabolite production in submerged mycelial processes. Biotechnol Adv 22:189–259
Park YS, Ohta N, Okabe M (1993) Effect of dissolved oxygen concentration and impeller tip speed on itaconic acid production by Aspersillus terreus. Biotechnol Lett 15:583–586
Paul BD, Snyder SH (2010) The unusual amino acid l-ergothioneine is a physiologic cytoprotectant. Cell Death Differ 17:1134–1140
Prosser JI, Tough AJ (1991) Growth mechanisms and growth kinetics of filamentous microorganisms. Crit Rev Biotechnol 10:253–274
Rahman I, Gilmour PS, Jimenez LA, Biswas SK, Antonicelli F, Aruoma OI (2003) Ergothioneine inhibits oxidative stress- and TNF-α-induced NF-κB activation and interleukin-8 release in alveolar epithelial cells. Biochem Biophys Res Commun 302:860–864
Reichl U, King R, Gilles ED (1992) Characterization of pellet morphology during submerged growth of Streptomyces tendae by image analysis. Biotechnol Bioeng 39:164–170
Riley GL, Tucker KG, Paul GC, Thomas CR (2000) Effect of biomass concentration and mycelial morphology on fermentation broth rheology. Biotechnol Bioeng 68:160–172
Riscaldati E, Moresi M, Federici F, Petruccioli M (2000) Effect of pH and stirring rate on itaconate production by Aspergillus terreus. J Biotechnol 83:219–230
Rühl M (2009) Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus—submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases. Ph.D dissertation. Georg-August-University Göttingen, Göttingen
Schügerl K, Wittler R, Lorenz T (1983) The use of molds in pellet form. Trends Biotechnol 1:120–123
Sinha J, Bae JT, Park JP, Kim KH, Song CH, Yun JW (2001) Changes in morphology of Paecilomyces japonica and their effect on broth rheology during production of exo-biopolymers. Appl Microbiol Biotechnol 56:88–92
Sobczuk TM, Camacho FG, Grima EM, Chirti Y (2005) Effects of agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. Bioprocess Biosyst Eng 28:243–250
Tang YJ, Zhong JJ (2003) Role of oxygen supply in submerged fermentation of Ganoderma lucidum for production of Ganoderma polysaccharide and ganoderic acid. Enzyme Microb Technol 32:478–484
Turło J, Gutkowskaa B, Herold F, Krzyczkowski W, Błazewicz A, Kocjan R (2008) Optimizing vitamin B12 biosynthesis by mycelial cultures of Lentinula edodes (Berk.) Pegl. Enzyme Microb Technol 43:369–374
Wagner R, Mitchell DA, Sassaki GL, Amazonas MALA (2004) Links between morphology and physiology of Ganoderma lucidum in submerged culture for the production of exopolysaccharide. J Biotechnol 114:153–164
Yang FC, Liau CB (1998) The influence of environmental conditions on polysaccharide formation by Ganoderma lucidum in submerged cultures. Process Biochem 33:547–553
Yang YK, Sbimizu H, Shioya S, Suga K, Nihira T, Yamada Y (1995) Optimum autoregulator addition strategy for maximum virginiamycin production in batch culture of Streptomyces virginiae. Biotechnol Bioeng 46:437–442
Zhong JJ, Yue CJ (2005) Plant cells: secondary metabolite heterogeneity and its manipulation. Biotechnology for the future. Adv Biochem Eng Biotechnol 100:53–88
Zorn H, Bouws H, Takenberg M, Nimtz M, Getzlaff R, Breithaupt DE, Berger RG (2005) An extracellular carboxylesterase from the basidiomycete Pleurotus sapidus hydrolyses xanthophyll esters. Biol Chem 386:435–440
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Tepwong, P., Giri, A. & Ohshima, T. Effect of mycelial morphology on ergothioneine production during liquid fermentation of Lentinula edodes . Mycoscience 53, 102–112 (2012). https://doi.org/10.1007/s10267-011-0145-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10267-011-0145-0