The effect of citrate and sulfate of copper on the biomass composition of the medicinal mushroom Trametes versicolor (Polyporales, Polyporaceae)

  • G. A. Al-Maali Kholodny Institute of botany NAS of Ukraine
  • N. A. Bisko Kholodny Institute of botany NAS of Ukraine
  • A. M. Ostapchuk Zabolotny Institute of Microbiology and Virology NAS of Ukraine
Keywords: lipids, crude protein, amino acids, fatty acids


The aim of our research was to study the influence of citrate and sulfate of copper on the biomass composition of the mycelium of the medicinal mushroom Trametes versicolor (L.) Lloyd, (1920) cultivated in a liquid medium. The studied strain of Trametes versicolor 353 was obtained from the Culture Collection of Mushrooms (IBK) from M.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine. Copper citrate was obtained from the Institute of Nanobiotechnologies and Resource Conservation of Ukraine, Kyiv. In this study we used glucose-peptone-yeast extract medium. Cu2+ (sulfate or citrate form) was added to the medium in concentration 4 mg/L. Mycelium was grown in a submerged culture on a rotary shaker (120 rpm) at 26 °C in 250 ml Erlenmeyer flasks, containing 50 ml of liquid media. The biomass was harvested after 9 days of cultivation in the liquid medium, filtered, washed, dried to a constant weight at 105 °C and weighed. Total nitrogen content (Ntotal) in the mycelium determined by the Kjeldahl method, crude protein content was determined as Ntotal x 6.25. The ash was obtained by the standard method. Total lipids were extracted from undried mycelium by a modified method of Bligh and Dyer. Amino acid composition was analyzed by high-performance liquid chromatography Agilent 1200 (Agilent technologies, USA). The methyl ethers of fatty acids were determined by gas chromatography-mass spectrometry (GC/MS) Agilent 6890N/5973 inert. The results of our research demonstrated that sulfates and citrates of copper increased the amount of crude protein on the mycelium of T. versicolor 353. Also both form of copper increased the amount of ash by a third relative to the control medium. At the same time, both forms of copper reduced the amount of total carbohydrates on mycelial biomass. But copper sulfate reduced the amount of total lipids relative to the control medium and medium with copper citrate. It should be noted, that given the significant growth of biomass in both cases, the yield of the same biomass compounds (gram per liter of medium) was raised relative to the control medium. So the yield of total carbohydrates was increased by 72% (on the medium with copper citrate) and 43% (on the medium with copper sulfate) relative to the control medium. The yield of crude protein was raised by 94% (on the medium with copper citrate) and 63% (on the medium with copper sulfate) relative to the control medium. Assay of amino acid composition showed that the quality of crude protein didn’t change. Thus the yield of essential amino acids was increased in conjunction with the yield of crude protein. The yield of total lipids increased only on the medium with copper citrate (by 57%) and in this case the content of fatty acids was unchanged significantly relative to the control medium. But sulfate of copper decreased the amount of cis-linoleic acids by 7%, in return the amount of oleic acid was increased relative to the control medium without copper. 


Al-Maali, G.A., 2015. The influence of metal citrates obtained by aquananotechnology on growth of the strains of medical macromycetes Ganoderma lucidum 1900 and Trametes versicolor 353. Ukrainian Botanical Journal 72(4), 393–397.

Antonenko, L.A., 2013. Biotekhnolohiia otrymmannia biomasy vyshchykh bazydialnykh hrybiv rodu Coriolus [Biotechnology of biomass higher basidiomycetes of the genus Coriolus]. NUHT, Kyiv (in Ukrainian).

Banci, L., 2013. Metallomics and the Cell. Springer, Dordrecht.

Bandura, I.I., 2014. Udoskonalennja elementiv tehnologii’ promyslovogo vyrobnyctva i’stivnyh grybiv rodu Pleurotus (Fr.) P. Kumm. [Improvement of technological elements for industrial production of edible mushroom Pleurotus (Fr.) P. Kumm.]. Edelveis, Kyiv (in Ukrainian).

Barna, M.M., 1997. Botanika. Terminy. Poniattia. Personalii [Botany. Terms. Concepts. Personality]. Akademia, Kyiv (in Ukrainian).

Borysevych, V.B., Kaplunenko, V.H., 2010. Nanomaterialy v biolohii. Osnovy nanoveterynarii [Nanomaterials in biology. Fundamentals of nanoveterinary medicine]. Avicena, Kyiv (in Ukrainian).

Buchalo, A.S., Mytropolska, N.Y., Mykchaylova, O.B., 2011. Catalogue of the culture collection of mushrooms IBK. Alterpress, Kiev.

Cai, X., Pi, Y., Zhou, X., Tian, L., Qiao, S., Lin, J., 2010. Hepatoma cell growth inhibition by inducing apoptosis with polysaccharide isolated from Turkey tail medicinal mush-room, Trametes versicolor (L.: Fr.) Lloyd (Aphyllophoromycetideae). Int. J. Med. Mushrooms 12(3), 257–263.

Christie, W.W., 1989. Gas chromatography and lipids: A practical guide. Oily Press Ltd, Ayr, Scotland.

Cunniff, P., 1995. AOAC Official methods of analysis (16th ed.). Association of Official Analytical Chemists, Arlighto.

Fedotov, O.V., Chayka, A.V., 2015. Destrukcija ksenobiotykiv z vykorystannjam kul'tural'nogo fil'tratu ksylotrofiv [Destruction of xenobiotics by culture filtrate from xylotrophic Basidiomycetes]. Biological Bulletin of Bogdan Chmelnitskiy Melitopol State Pedagogical University 5(3), 55–72 (in Ukrainian).

Gralla, E.B., Thiele, D.J., Silar, P., Valentine, J.S., 1991. ACE1, a copper-dependent transcription factor, activates expression of the yeast copper, zinc superoxide dismutase gene. Proc. Natl. Acad. Sci. USA 88(19), 8558–8562.

Henderson, J.W., Ricker, R.D., Bidlingmeyer, B.A., Woodward, C., 2000. Rapid, accurate, sensitive, and reproducible HPLC analysis of amino. Agilent Technologies, Technical Note 5980–1193E.

Ivanova, T.S., Bisko, N.A., Barshteyn, V.J., Krupodorova, T.A., 2010. Biologichno-aktyvni rechovyny grybiv viddilu Basidiomycota [Biologically active substances of fungi Basidiomycota]. Problemy Harchuvannja 1(2), 42–47.

Jámbor, A., Molnár-Perl, I., 2009a. Amino acid analysis by high-performance liquid chromatography after derivatization with 9-fluorenylmethyloxycarbonyl chloride. Literature overview and further study. J. Chromatogr. A 1216, 3064–3077.

Jámbor, A., Molnár-Perl, I., 2009b. Quantitation of amino acids in plasma by high performance liquid chromatography: Simultaneous deproteinization and derivatization with 9-fluorenylmethyloxycarbonyl chloride. J. Chromatogr. A 1216, 6218–6223.

Kroneck, P.M.H., Sosa Torres M.E. (eds.), 2015. Sustaining life on planet earth: Metalloenzymes mastering dioxygen and other chewy gases. Springer International Publishing, Switzerland.

Kuan, Y.C., Wu, Y.J., Hung, C.L., Sheu, F., 2013. Trametes versicolor protein YZP activates regulatory B lymphocytes–gene identification through de novo assembly and function analysis in a murine acute colitis model. PLoS ONE 8(9), e72422.

Maehara, Y., Tsujitani, S., Saeki, H., Oki, E., Yoshinaga, K., Emi, Y., Baba, H., 2012. Biological mechanism and clinical effect of protein-bound polysaccharide K (Krestin®): Review of development and future perspectives. Surg. Today 42(1), 8–28.

Manirakiza, P., Covaci, A., Schepens, P., 2001. Comparative study on total lipid determination using soxhlet, roese-gottlieb, bligh and dyer, and modified bligh and dyer extraction methods. J. Food Compos. Anal. 14, 93–100.

Patel, S., Goyal, A., 2012. Recent developments in mushrooms as anticancer therapeutics: A review. 3 Biotech. 2(1), 1–15.

Sejfulla, R.D., Rozhkova, E.A., Kim, E.K., 2009. Antioksidanty [Antioxidants]. Jeksperimental’naja i Klinicheskaja Farmakologija 72(3), 60–64.

Soden, D.M., Dobson, A.D., 2001. Differential regulation of laccase gene expression in Pleurotus sajor-caju. Microbiology 147(7), 1755–1763.

Standish, L.J., Wenner, C.A., Sweet, E.S., Bridge, C., Nelson, A., Martzen, M., Torkelson, C., 2008. Trametes versicolor mushroom immune therapy in breast cancer. J. Soc. Integr. Oncol. 6(3), 122–128.

Vasina, D.V., Mustafaev, O.N., Moiseenko, K.V., Sadovskaya, N.S., Glazunova, O.A., Tyurin, А.А., Koroleva, O.V., 2015. The Trametes hirsuta 072 laccase multigene family: Genes identification and transcriptional analysis under copper ions induction. Biochimie 116, 154–164.

Xiao, J.H., Chen, D.X., Wan, W.H., Hu, X.J., Qi, Y., Liang, Z.Q., 2006. Enhanced simultaneous production of mycelia and intracellular polysaccharide in submerged cultivation of Cordyceps jiangxiensis using desirability functions. Process Biochem. 41(8), 1887–1893.

Zhi-Ling, C., 2009. Effect of some trace elements and vitamins on contents of polysaccharide and acid of Ganoderma lucidum. Journal of Anhui Agricultural Sciences 5, 078.

Zong, A., Cao, H., Wang, F., 2012. Anticancer polysaccharides from natural resources: A review of recent research. Carbohydr. Polym. 90(4), 1395–1410.

Zou, X., 2005. Effects of Zn supplementation on the growth, amino acid composition, polysaccharide yields and anti-tumour activity of Agaricus brasiliensis. World J. Microbiol. Biotechnol. 21(3), 261–264.