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Optimized Culture Conditions for Enhanced Recovery of Exopolysaccharide from Pseudolagarobasidium acaciicola:A Novel Fungus Isolated from the Fruit Body of Russula nigricans, A Wild Edible Mushroom of Odisha, India


Affiliations
1 Plant Pathology and Microbiology Division, Regional Plant Resource Centre, Bhubaneswar 751 015, India
 

Fungal exopolysaccharides (EPS) are becoming important due to their multifarious applications with different structural forms and easy recovery. The objective of this study was to optimize submerged culture condition of a new fungal isolate Pseudolagarobasidium acaciicola obtained from fruit body of an edible mushroom, Russula nigricans. The study analyses the optimization of different parameters for enhanced production of EPS by one factor-at-a-time (OFAT) method. The influence of incubation period, initial pH value, temperature, mode of culture (static shake), culture vessel, carbon and nitrogen sources, and enhancers was studied. OFAT method revealed pH 6 with 7 days incubation statically and in dark in 150 ml Erlenmeyer flask, chemical factors like sabouraud dextrose HiVeg broth medium, xylose, yeast extract, tryptophan, K2HPO4, CaCl2 and vitamin C as good conditions and components for maximum biomass and EPS production. Optimized medium developed in this study was a combination of the individually screened nutrient component, estimated the maximum EPS (1002.3 ± 189.72 mg/l) which was later expelled to 1468.1 ± 227.86 mg/l after addition of olive oil and Tween 80 at a concentration 250 : 50 μl v/v, which was much higher, and reported first time from this fungus (it means that early when medium was formulated with different chemical components we got the optimized medium giving 1002.3 mg/l of EPS but when addition of oils was performed we got more amount, i.e. 1468.1 mg/l). EPS production in a new medium might facilitate its industrial-scale production and use as a bioactive product for the welfare of mankind.

Keywords

Exopolysaccharide, Optimization, Pseudolagarobasidium acaciicola, Submerged Culture.
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  • Ugwu, E. E. and Adebayo-Tayo, B. C., Screening of some Basidiomycetes for bio-polymers and biomass production in submerged cultivation. AUJT, 2011, 15, 41–44.
  • Ahmed, S., Anwar, A., Haider, A., Adnan Saeed, M., Nadeem, M., Zahida Nasreen, Z. and Baig, S., Selection of culture medium for exopolysaccharides production by Coriolus versicolor. Pak. J. Phytopathol., 2011, 23, 1–4.
  • Abdel-Aziz, S. M., Hamed, H. A., Mouafi, F. E. and Gad, A. S., Acidic pH-shock induces the production of an exopolysaccharide by the fungus Mucor rouxii: Utilization of beet-molasses. NY Sci. J., 2012, 5, 52–61.
  • Looijestenijn, P. J., Boels, I. C., Kleerebenzem, M. and Hugenholtz, J., Regulation of the exopolysaccharide production by Lactococcus lactis subsp. cremoris by the sugar source. Appl. Environ. Microbiol., 1999, 65, 5003–5008.
  • Levander, F. and Radstrom, P., Requirement for phosphoglucomutase in exopolysaccharide biosynthesis in glucose and lactose utilizing Streptococcus thermophilus. Appl. Environ. Microbiol., 2001, 67, 2734–2738.
  • Richert, L., Golubic, S., Guedes Ratiskol, J. and Payri, C., Characterization of exopolysaccharides produced by cyanobacteria isolated from Polynesian microbial mats. Curr. Microbiol., 2005, 25, 379–384.
  • Miranda, B. G. and Leal, J. A., Extracellular and cell wall polysaccharides of Aspergillus alliaceus. Trans. Br. Mycol. Soc., 1981, 76, 249–253.
  • Stasinopoulos, S. J. and Seviour, R. J., Exopolysaccharide formation by isolates of Cephalosporium and Acremonium. Mycol. Res., 1989, 92, 55–60.
  • Graber, M., Morin, A., Duchiron, F. and Monsan, P. F., Microbial polysaccharides containing 6-deoxysugars. Enzyme Microb, Technol., 1988, 10, 198–206.
  • Nehad, E. A. and Shamy, A. R. I., Optimization of polysaccharide production by Alternaria alternata. Gate2Biotech, 2010, 1, 1–6.
  • Li, P., Luo, C., Sun, W., Lu, S., Mou, Y., Peng, Y. and Zhou, L., In vitro antioxidant activities of polysaccharides from endophytic fungus Fusarium oxysporum. Afr. J. Microbiol. Res., 2011, 5.
  • Uyanoglu, M., Canbek, M., Ozalp, F. O., Yamac, M. and Senturk, H., Effects of some macrofungi exopolysaccharides on mesenchymal mast cells of rats in chronic alcohol consumption. Int. J. Health Nutr., 2010, 1, 31–37.
  • Su, C. et al., Isolation and characterization of exopolysaccharide with immunomodulatory activity from fermentation broth of Morchella conica. DARU J. Pharma. Sci., 2013, 21, 1–6.
  • Chiu, Y. W., Zeng, C. L., Chian, P. L. and Shiu, H. W., Effect of carbon and nitrogen sources on the production and carbohydrate composition of exopolysaccharides by submerged culture of Pleurotus citrinopileatus. J. Food Drug Anal., 2008, 16, 61–67.
  • Cheung, P. C. K., The hypocholesterolemic effect of extracellular polysaccharide from the submerged fermentation of mushroom. Nutr. Res., 1996, 16, 1953–1957.
  • Borchers, A. T., Stern, J. S., Hackman, R. M., Keen, C. L. and Gershwin, M. E., Mushrooms, tumors, and immunity. Proc. Soc. Biol. Med., 1999, 221, 281–293.
  • Yang, B. K. et al., Production of exo-polymers by submerged mycelial culture of Cordyceps militaris and its hypolipidemic effect. J. Ind. Microbiol. Biotechnol., 2000, 10, 784–788.
  • Cohen, R., Persky, L. and Hadar, Y., Biotechnological applications and potential of wood-degrading mushroom of the genus Pleurotus. Appl. Microbiol. Biotechnol., 2002, 58, 582–594.
  • Hwang, H. J., Kim, S. W., Xu, C. P., Choi, J. W. and Yun, J. W., Morphological and rheological properties of the three different species of basidiomycetes Phellinus in submerged cultures. J. Appl. Microbiol., 2004, 96, 1296–1305.
  • Ross, P. R. et al., Developing applications for lactococcal bacteriocins. Antonie van Leeuwenhoek, 1999, 76, 337–346.
  • Lee, K. Y., Lee, M. H., Chang, L. Y., Yoon, S. P., Lim, D. Y. and Jeon, Y. J., Macrophage activation by polysaccharide fraction isolated from Salicornia herbacea. J. Ethnopharmacol., 2006, 103, 372–378.
  • Paulraj, B. and Saravanan, T., Optimization of Β-glucan production from lower fungi using central composite design and its biological application. Int. J. Comput. Appl., 2012, 49, 23–28.
  • Kanari, B., Banik, R. R. and Upadhyay, S. N., Effect of environmental factors and carbohydrate on gellan gum production. Appl. Biochem. Biotechnol., 2002, 102–103(1–6), 129–140.
  • Kim, S. W., Hwang, H. J., Park, J. P., Cho, Y. J., Song, C. H. and Yun, J. W., Mycelial growth and exo-biopolymer production by submerged culture of various edible mushrooms under different media. Lett. Appl. Microbiol., 2002, 34, 56–61.
  • Tang, Y. J. and Zhong, J. J., Exopolysaccharide biosynthesis and related enzyme activities of the medicinal fungus, Ganoderma lucidum, grown on lactose in a bioreactor. Biotechnol. Lett., 2002, 24, 1023–1026.
  • Kwon, J. S., Lee, J. S., Shin, W. C., Lee, K. E. and Hong, E. K., Optimization of culture conditions and medium components for the production of mycelial biomass and exo-polysaccharides with Cordyceps militaris in liquid culture. Biotechnol. Bioprocess Eng., 2009, 14, 756–762.
  • Hwang, H. S. and Yun, J. W., Hypogycemic effect of polysaccharides produced by submerged mycelia culture of Laetiporus sulphureus on streptozotocin induced diabetics rats. Biotechnol. Bioprocess Eng., 2010, 15, 173–181.
  • Kim, S. S., Lee, J. S., Cho, J. Y., Kim, Y. E. and Hong, E. K., Effects of C/N ratio and trace elements on mycelial growth and exo-polysaccharide production of Tricholoma matsutake. Biotechnol Bioprocess Eng., 2010, 15, 293–298.
  • Anandapandian, K. T. K. and Eyini, M., Optimization of mycelial growth and exopolysaccharide production by Calocybe indica using response surface methodology. J. Adv. Med. Life Sci., 2014, 1, 1–8.
  • Silvi, S., Barghini, P., Aquilanti, A., Juarez-Jimenez, B. and Fenice, M., Physiologic and metabolic characterization of a new marine isolate (BM39) of Pantoea sp. producing high levels of exopolysaccharide. Microbial. Cell Fact., 2013, 12, 1–11.
  • Kim, S. W., Xu, C. P., Hwang, H. J., Choi, J. W., Kim, C. W. and Yun, J. W., Production and characterization of exopolysaccharides from an enthomopathogenic fungus Cordyceps militaris NG3. Biotechnol. Prog., 2003, 19, 428–435.
  • Banerjee, D., Jana, M. and Mahapatra, S., Production of exopolysaccharide by endophytic Stemphylium sp. Micol. Apl. Int., 2009, 21, 57–62.
  • Hamedi, A., Vahid, H. and Ghanati, F., Optimization of medium composition for production of mycelia biomass and exopolysaccharide by Agaricus blazei Murill DPPH 131 using response surface methodology. Biotechnology, 2007, 6, 456–464.
  • Audy, J., Labrie, S., Roy, D. and LaPointe, G., Sugar source modulates exopolysaccharide biosynthesis in Bifidobacterium longum subsp. longum CRC 002. Microbiology, 2010, 156, 653–664.
  • Joshi, M., Patel, H., Gupte, S. and Gupte, A., Nutrient improvement for simultaneous Schizophyllum commune AGMJ-1 using statistical optimization. 3 Biotech, 2013, 3, 307–318.
  • Li, R., Jiang, X. and Guan, H., Optimization of mycelium biomass and exopolysaccharides production by Hirsutella sp. in submerged fermentation and evaluation of exopolysaccharides antibacterial activity. Afr. J. Biotechnol., 2010, 9, 195–202.
  • Ma, Y. P., Mao, D. B., Geng, L. J., Zhang, W. Y., Wang, Z. and Xub, C. P., Optimization, molecular characterization and biological activities of exopolysaccharides from Xylaria nigripes. Chem. Biochem. Eng., 2013, 27, 177–184.
  • Lim, J. M., Kim, S. W., Hwang, H. J., Joo, J. H., Kim, H. O., Choi, J. W. and Yun, J. W., Optimization of medium by orthogonal matrix method for submerged mycelial culture and exopolysaccharide production in Collybia maculate. Appl. Biochem. Biotechnol., 2004, 119, 159–170.
  • Kaur, V., Bera, M. B., Panesar, P. S. and Chopra, H. K., Production and characterization of exopolysaccharide produced by Alcaligenes faecalis B14 isolated from indigenous soil. Int. J. Biotechnol. Bioeng. Res., 2013, 4, 365–374.
  • Shen, J., Shi, C. and Xu, C., Exopolysaccharides from Pleurotus pulmonarius: fermentation optimization, characterization and antioxidant activity. Food Technol. Biotechnol., 2013, 51, 520– 527.
  • Ko, S., Lee, H. S., Park, S. H. and Lee, H. K., Optimal conditions for the production of exopolysaccharide by marine microorganism Hahella chejuensis. Biotechnol. Bioprocess Eng., 2000, 5, 181–185.
  • Adebayo-Tayo, B. C., Jonathan, S. G., Popoola, O. O. and Egbomuche, R., Optimization of growth conditions for mycelia yield and exopolysaccharide production by Pleurotus ostreatus cultivated in Nigeria. Afr. J. Microbiol. Res., 2011, 5, 2130–2138.
  • Torino, M. I., Hebert, E. M., Mozzi, F. and Font de Valdez, G., Growth and exopolysaccharide production by Lactobacillus helveticus ATCC 15807 in an adenine-supplemented chemically defined medium. J. Appl. Microbiol., 2005, 99, 1123–1129.
  • Silva, C. C., Robert, F. H., Dekker Silva, R. S. S. F., da Silva, M. L. C. and Aneli Barbosa, M., Effect of soybean oil and Tween 80 on the production of botryosphaeran by Botryosphaeria rhodina MAMB-05. Process Biochem., 2007, 42, 1254–1258.
  • Halim, K. H. A., Kamal, I. S. M., Rashid, N. M. N. and Maizirwan, M., BPE-P04: the effect of plant oils for submerged fermentation of Schizophyllum commune producing mycelium biomass and exopolysaccharides. In Malaysian International Conference on Trends in Bioprocess Engineering (MICOTriBE), Langkawi, Malaysia, 2012, pp. 1–7.
  • Qing, H. F. and Jian, J. Z., Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum. Process Biochem., 2002, 37, 769–774.
  • Lv, Y. L., Sun, L. H., Zhang, F. S., Zhoa, Y. and Guo, S. X., The effect of cultivation conditions on the mycelia growth of a dark-septate endophytic isolate. Afr. J. Microbiol. Res., 2010, 4, 602– 607.
  • Xiao, J. H. et al., Optimization of submerged culture requirements for the production of mycelial growth and exopolysaccharide by Cordyceps jiangxiensis JXPJ 0109. J. Appl. Microbiol., 2004, 96, 1105–1116.
  • Maziero, R., Cavazzoni, V. and Bononi, V. L. R., Screening of basidiomycetes for the production of exopolysaccharide and biomass in submerged culture. Rev. Microbiol., 1999, 30, 77–84.
  • Shih, I. L., Pan, K. and Hsieh, C., Influence of nutritional components and oxygen supply on the mycelial growth and bioactive metabolites production in submerged culture of Antrodia cinnamomea. Process Biochem., 2006, 41, 1129–1135.
  • Ahmed, R., Al-Shorgani, N. K. N., Hamid, A. A., Yusoff, W. M. W. and Daud, F., Optimization of medium components using response surface methodology (RSM) for mycelium biomass and exopolysaccharide production by Lentinus squarrosulus. Adv. Biosci. Biotechnol., 2013, 4, 1079–1085.
  • Dubois, M., Gilles, Gilles, K. A., Hamilton, J. K., Rabers, P. A. and Smith, F., Colorimetric method for determination of sugars and related substances. Anal. Chem., 1956, 28, 350–356.
  • Bolla, K., Shaheen, S. Z., Vasu, K. and Charya, M. A. S., Effect of oils on the production of exopolysaccharides and mycelia biomass in submerged culture of Schizophyllum commune. Afr. J. Microbiol. Res., 2008, 2, 349–352.
  • Bolla, K., Hima Bindu, N. S. V. S.S .S. L., Burra, S. and Charya, M. A. S., Effect of plant oils, surfactant and organic acids on the production of mycelia biomass and exopolysaccharides of Trametes spp. J. Agric. Technol., 2011, 7, 957–965.
  • Shankar, T., Vijayabaskar, P., Narayani, S. and Sivakumar, T., Screening of exopolysaccharide producing bacterium Frateuria aurentia from elephant dung. Appl. Sci. Rep., 2014, 1, 105–109.
  • Smiderle, F. R. et al., Exopolysaccharides, proteins and lipids in Pleurotus pulmonarius submerged culture using different carbon sources. Carbohydr. Polym., 2012, 87, 368–376.
  • Zhou, Q., Yang, W., Lin, J. and Guo, L., Optimization of medium pH, growth media compositions and analysis of nutritional components of Ganoderma lucidum in submerged culture fermentation. Eur. J. Med. Plants, 2015, 6, 17–25.
  • Kim, H. O. and Yun, J. W., A comparative study on the production of exopolysaccharides between two enthomopathogenic fungi Cordyceps militaris and Cordyceps sinensis in submerged mycelial cultures. J. Appl. Microbiol., 2005, 99, 728–738.
  • Thakur, S. and Gupte, A., Optimization and hyper production of laccase from novel agaricomycete Pseudolagarobasidium acaciicola AGST3 and its application in in vitro decolorization of dyes. Ann. Microbiol., 2015, 65, 185–196.
  • Adak, R., Tiwari, R., Sing, S. and Nain, L., Laccase production by a novel white-rot fungus Pseudolagarobasidium acaciicola LA1 through solid state fermentation of Parthenium biomass and its application in dyes decolorization. Waste Biomass Valori., 2016, 7.10.1007/s12649-016-9550.0.
  • Wibowo, M., Prachyawarakorn, R., Aree, T., Mahidol, C., Ruchirawat, S. and Kittakoo, P., Cytotoxic sesquiterpenes from the endophytic fungus Pseudolagarobasidium acaciicola. Phytochemisry, 2016, 122, 126–138.
  • Mahmoud Ei-Dein, M. N., Amira EI-Fallal, A., El-Shahat Toson, A. and Faten Hereher, E., Exopolysaccharides production by Pleurotus pulmonarius: Factors affecting formation and their structures. Pak. J. Biol. Sci., 2004, 7, 1078–1084.
  • Adebayo-Tayo, B. C. and Ugwu, E. E., Influence of different nutrient sources on exopolysaccharide production and biomass yield by submerged culture of Trametes versicolor and Coprinus sp. AUJT, 2011, 15, 63–69.
  • Patil, S. V., Patil, C. D., Salunke, R. B., Bathe, G. A. and Patil, D. M., Studies on characterization of bioflocculant exopolysaccharide of Azotobacter indicus and its potential for wastewater treatment. Appl. Biochem. Biotechnol., 2011, 163, 463–472.
  • Zhang, J. et al., Extraction optimization of exopolysaccharide produced by Pleurotus cornucopiae SS-02 and its antioxidant activity. Afr. J. Biotechnol., 2012, 11, 4815–4825.
  • Saskiawan, I., Exopolysaccharide production and its bioactivities of the edible Pleurotus ostreatus in submerged culture. Biotropia, 2009, 16, 96–104.
  • Prathumpai, W., Rachathewee, P., Khajeeram, S., Sanglier, J., Tanjak, P. and Methacanon, P., Optimization, characterization and in vitro evaluation of entomopathogenic fungal exopolysaccharides as prebiotic. Adv. Biochem., 2013, 1, 13–21.
  • Hwang, D. et al., Nutritional requirements for the mycelial biomass and exopolymer production by Hericium erinaceus CZ-2. Food Technol. Biotechnol., 2007, 45, 389–395.
  • Liu, Y.-S., and Wu, J.-Y., Effects of Tween 80 and pH on mycelial pellets and exopolysaccharide production in liquid culture of a medicinal fungus. J. Ind. Microbiol. Biotechnol., 2012, 39, 623–628.
  • Hao, M., Xing, X. H., Li, Z., Zhang, J. C., Sun, J. X., Qiao, C. S. and Wu, T., Optimization of effect factors for mycelia growth and exopolysaccharide production by Schizophyllum commune. Appl. Biochem. Biotechnol., 2010, 160, 621–631.
  • Li, Y., Guo, S. and Zhu, H., Statistical optimization of culture medium for production of exopolysaccharide from endophytic fungus Bionectria ochroleuca and its antitumor effect in vitro. EXCLI-J. 2016, 15, 211–250.
  • Mshandate, A. M. and Mgonia, J., Submerged liquid fermentation of some Tanzanian basidiomycetes for the production of mycelia biomass, exopolysaccharides and mycelium protein using wastes peels media. ARPN J. Agri. Biol. Sci., 2009, 4, 1–13.
  • Hsieh, C., Wang, H. L., Chen, C. C., Hsu, T. H. and Tseng, M. H., Effect of plant oil and surfactant on the production of mycelial biomass and polysaccharides in submerged culture of Grifola frondosa. Biochem. Eng. J., 2008, 38, 198–205.
  • Li, R., Jiang, X. and Guan, H., Optimization of mycelium biomass and exopolysaccharides produced by Hirsutella sp.in submerged fermentation and evaluation of exopolysaccharides antibacterial activity. Afr. J. Biotechnol., 2010, 9, 195–202.
  • Burns, P. J., Yeo, P., Keshavarz, T., Roller, S. and Evans, C. S., Physiological studies of exopolysaccharide production from the basidiomycete Pleurotus sp. Florida. Enzyme Microb. Technol., 1994, 16, 566–572.
  • Kim, S. S., Lee, J. S., Cho, J. Y., Kim, Y. E. and Hong, E. K., Effects of C/N ratio and trace elements on mycelial growth and exopolysaccharide production of Tricholoma matsutake. Biotechnol. Bioprocess Eng., 2010, 15, 293–298.
  • Kim, H. H., Na, J. G., Chang, Y. K., Chun, G. T., Lee, S. J. and Jeong, Y. H., Optimization of submerged culture conditions for mycelia growth and exopolysaccharides production by Agaricus blazei. J. Microbiol.. Biotechnol., 2004, 14, 944–951.
  • Wang, F., Zhang, J., Hao, L., Jia, S., Ba, J. and Niu, S., Optimization of submerged culture conditions for mycelial growth and extracellular polysaccharide production by Coriolus versiolor. J. Bioproces. Biotech., 2012, 2, 124; doi:10.4172/2155-9821.1000124.
  • Adebayo-Tayo, B. C., Ugwu, E. E. and Musa, H., Physiological requirement for growth and extracellular polysaccharides (EPS) production by Marasmius sp. and Fomes sp. (a comparative study). J. Microbiol. Biotech. Res., 2013, 3, 1–11.
  • Todorov, S. D. and Dicks, L. M., Bacteriocin production by Pediococcus pentosaceus isolated from marula (Scerocarya birrea). Int. J. Food Microbiol., 2009, 132, 117–126.
  • Ogunbanwo, S., Sanni, A. and Onilude, A., Influence of culture conditions on the production of bacteriocin by Lactobacillus brevis OGI. Afr. J. Biotechnol., 2003, 7, 179–184.

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  • Optimized Culture Conditions for Enhanced Recovery of Exopolysaccharide from Pseudolagarobasidium acaciicola:A Novel Fungus Isolated from the Fruit Body of Russula nigricans, A Wild Edible Mushroom of Odisha, India

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Authors

Smita Behera
Plant Pathology and Microbiology Division, Regional Plant Resource Centre, Bhubaneswar 751 015, India
Nibha Gupta
Plant Pathology and Microbiology Division, Regional Plant Resource Centre, Bhubaneswar 751 015, India

Abstract


Fungal exopolysaccharides (EPS) are becoming important due to their multifarious applications with different structural forms and easy recovery. The objective of this study was to optimize submerged culture condition of a new fungal isolate Pseudolagarobasidium acaciicola obtained from fruit body of an edible mushroom, Russula nigricans. The study analyses the optimization of different parameters for enhanced production of EPS by one factor-at-a-time (OFAT) method. The influence of incubation period, initial pH value, temperature, mode of culture (static shake), culture vessel, carbon and nitrogen sources, and enhancers was studied. OFAT method revealed pH 6 with 7 days incubation statically and in dark in 150 ml Erlenmeyer flask, chemical factors like sabouraud dextrose HiVeg broth medium, xylose, yeast extract, tryptophan, K2HPO4, CaCl2 and vitamin C as good conditions and components for maximum biomass and EPS production. Optimized medium developed in this study was a combination of the individually screened nutrient component, estimated the maximum EPS (1002.3 ± 189.72 mg/l) which was later expelled to 1468.1 ± 227.86 mg/l after addition of olive oil and Tween 80 at a concentration 250 : 50 μl v/v, which was much higher, and reported first time from this fungus (it means that early when medium was formulated with different chemical components we got the optimized medium giving 1002.3 mg/l of EPS but when addition of oils was performed we got more amount, i.e. 1468.1 mg/l). EPS production in a new medium might facilitate its industrial-scale production and use as a bioactive product for the welfare of mankind.

Keywords


Exopolysaccharide, Optimization, Pseudolagarobasidium acaciicola, Submerged Culture.

References





DOI: https://doi.org/10.18520/cs%2Fv116%2Fi8%2F1397-1406