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Optimization of submerged culture conditions involving a developed fine powder solid seed for exopolysaccharide production by the medicinal mushroom Ganoderma lucidum

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Abstract

To facilitate Ganoderma lucidum submerged cultivation and achieve high productivity, four fine powder solid substrates incorporated with different nitrogen-rich supplements were utilized to grow the fungus and as solid seed for its submerged culture. Of the four solid seeds, the soybean meal solid seed gave the highest biomass (10.73 g/L) and exopolysaccharide (EPS) (1.22 g/L), higher than those (8.36 g/L biomass and 0.44 g/L EPS) obtained with mycelial liquid seed. The optimal level of soybean meal supplementation was 20% (w/w) for production of the solid seed. Following single factor experiments, levels of three selected process variables were optimized as: the moisture content of solid seed, 70%; inoculum size, 0.8 g/flask; and rotary speed, 160 rpm. These conditions were validated experimentally with improved EPS yield of 1.33 g/L. The developed solid seed can be conveniently used for G. lucidum submerged culture with improved EPS productivity.

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References

  • Bae JT, Sinha J, Park JP, Song CH, Yun JW. Optimization of submerged culture conditions for exo-biopolymer production by Paecilomyces japonica. J. Microbiol. Biotechnol. 10: 482–487 (2000)

    CAS  Google Scholar 

  • Box GP, Behnken DW. Some new three level design for the study of quantitative variables. Technometrics 2: 456–475 (1960)

    Article  Google Scholar 

  • Chang MY, Tsai GJ, Houng JY. Optimization of medium composition for the submerged culture of Ganoderma lucidum by Taguchi array design and steepest ascent method. Enzyme Microb. Technol. 38: 407–414 (2006)

    Article  CAS  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350–356 (1956)

    Article  CAS  Google Scholar 

  • Fang QH, Tang YJ, Zhong JJ. Significance of inoculation density control in production of polysaccharide and ganoderic acid by submerged culture of Ganoderma lucidum. Process Biochem. 37: 1375–1379 (2002)

    Article  CAS  Google Scholar 

  • Fang QH, Zhong JJ. Effect of initial pH on production of ganoderic acid and polysacchrides by submerged fermentation of Ganoderma lucidum. Process Biochem. 37: 769–774 (2002)

    Article  CAS  Google Scholar 

  • Feng J, Feng N, Zhang JS, Yang Y, Jia W, Lin CC. A new temperature control shifting strategy for enhanced triperpene production by Ganoderma lucidum G0119 based on submerged liquid fermentation. Appl. Biochem. Biotechnol. 180: 740–752 (2016)

    Article  CAS  PubMed  Google Scholar 

  • Feng J, Zhang JS, Feng N, Yan MQ, Yang Y, Jia W, Lin CC. A novel Ganoderma lucidum G0119 fermentation strategy for enhanced triterpenes production by statistical process optimization and addition of oleic acid. Eng. Life. Sci. 17: 430–439 (2017)

    Article  CAS  Google Scholar 

  • Gao CJ, Wang ZY, Su TT, Zhang J, Yang XH. Optimisation of exopolysaccharide production by Gomphidius rutilus and its antioxidant activities in vitro. Carbohydr. Polym. 87: 2299–2305 (2012)

    Article  CAS  Google Scholar 

  • Jia J, Yang XF, Wu ZL, Zhang Q, Lin Z, Guo HT, Lin CSK, Wang JY, Wang YS. Optimization of fermentation medium for extracellular lipase production from Aspergillus niger using response surface methodology. Biomed Res. Int. 2015: 497462 (2015)

  • Jong SC, Birmingham JM. Medicinal benefits of the mushroom Ganoderma. Adv. Appl. Microbiol. 37: 101–134 (1992)

    Article  CAS  PubMed  Google Scholar 

  • Kim HM, Kim SW, Hwang HJ, Park MK, Mahmoud YAG, Choi JW, Yun JW. Influence of agitation intensity and aeration rate on production of antioxidative exopolysaccharides from submerged mycelial culture of Ganoderma resinaceum. J. Microbiol. Biotechnol. 16: 1240–1247 (2006a)

    CAS  Google Scholar 

  • Kim HM, Park MK, Yun JW. Culture pH affects exopolysaccharide production in submerged mycelial culture of Ganoderma lucidum. Appl. Biochem. Biotechnol. 134: 249–262 (2006b)

    Article  CAS  PubMed  Google Scholar 

  • Le J, Hu SZ, Xu M. Optimization of submerged culture conditions for the production of mycelial biomass and exopolysaccharide by Pleurotus nebrodensis. Ann. Microbiol. 57: 389–393 (2007)

    Article  CAS  Google Scholar 

  • Lee KM, Lee SY, Lee HY. Biostage control of pH for improving exopolysaccharide production from mycelia of Ganoderma lucidum in an air-lift fermentor. J. Biosci. Bioeng. 88: 646–650 (1999)

    Article  CAS  PubMed  Google Scholar 

  • Li HX, Lu ZM, Geng Y, Gong JS, Zhang XJ, Shi JS, Xu ZH, Ma YH. Efficient production of bioactive metabolites from Antrodia camphorata ATCC 200183 by asexual reproduction-based repeated batch fermentation. Bioresour. Technol. 194: 334–343 (2015)

    Article  CAS  PubMed  Google Scholar 

  • Liu SR, Zhang WR. Hyperproduction of exopolysaccharides by submerged mycelial culture of Ganoderma lucidum using a solid seed grown in fine-powder of wheat bran and in vitro evaluation of the antioxidant activity of the exopolysaccharides produced. Food Sci. Biotechnol. 27: 1129–1136 (2018)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lonsane BK, Ghildyal NP, Budiatman S, Ramakrishna SV. Engineering aspects of solid state fermentation. Enzyme Microb. Technol. 7: 258–265 (1985)

    Article  CAS  Google Scholar 

  • Mizuno T, Wang GY, Zhang J, Kawagishi H, Nishitoba T, Ti JX. Reishi, Ganoderma lucidum and Ganoderma tsugae: bioactive substances and medicinal effects. Food Rev. Int. 11: 151–166 (1995)

    Article  CAS  Google Scholar 

  • Montgomery DC. Design and analysis of experiments (2nd ED). J. Am. Stat. Assoc. 16: 241–242 (2000)

    Google Scholar 

  • Moonmoon M, Shelly NJ, Khan MA, Uddin MN, Hossain K, Tania M, Ahmed S. Effects of different levels of wheat bran, rice bran and maize powder supplementation with saw dust on the production of shiitake mushroom (Lentinus edodes (Berk.) Singer). Saudi J. Biol. Sci. 18: 323–328 (2011)

    Article  PubMed  Google Scholar 

  • Naraian R, Sahu RK, Kumar S, Garg SK, Singh CS, Kanaujia RS. Influence of different nitrogen rich supplements during cultivation of Pleurotus florida on corn cob substrate. Environmentalist 29: 1–7 (2009)

    Article  Google Scholar 

  • Pandey A. Production of starch saccharifying enzyme (glucoamylase) in solid cultures. Starch 44: 75–77 (1992)

    Article  CAS  Google Scholar 

  • Postemsky PD, Delmastro SE, Curvetto NR. Effect of edible oils and Cu (II) on the biodegradation of rice by-products by Ganoderma lucidum mushroom. Int. Biodeter. Biodegr. 93: 25–32 (2014)

    Article  CAS  Google Scholar 

  • Reddy LVA, Wee YJ, Yun JS, Ryu HW. Optimization of alkaline protease production by batch culture of Bacillus sp. RKY3 through Plackett–Burman and response surface methodological approaches. Bioresour. Technol. 99: 2242–2249 (2008)

    Article  CAS  PubMed  Google Scholar 

  • Rocky-Salimi K, Hamidi-Esfahani Z. Evaluation of the effect of particle size, aeration rate and harvest time on the production of cellulase by Trichoderma reesei QM9414 using response surface methodology. Food Bioprod. Process. 88: 61–66 (2010)

    Article  CAS  Google Scholar 

  • Shiao MS. Natural products of the medicinal fungus Ganoderma lucidum: occurrence, biological activities, and pharmacological functions. Chem. Res. 3: 172–180 (2003)

    CAS  Google Scholar 

  • Shu CH, Lung MY. Effect of pH on the production and molecular weight distribution of exopolysaccharide by Antrodia camphorata in batch cultures. Process Biochem. 39: 931–937 (2004)

    Article  CAS  Google Scholar 

  • Song CH, Yang BK, Ra KS, Shon DH, Park EJ, Go GI, Kim YH. Hepatoprotective effect of extracellular polymer produced by submerged culture of Ganoderma lucidum WK-003. J. Microbiol. Biotechnol. 8: 277–279 (1998)

    Google Scholar 

  • Tepwong P, Giri A, Ohshima T. Effect of mycelial morphology on ergothioneine production during liquid fermentation of Lentinula edodes. Mycoscience 53: 102–112 (2012)

    Article  CAS  Google Scholar 

  • Upadhyay M, Shrivastava B, Jain A, Kidwai M, Kumar S, Gomes J, Goswami DG, Panda AK, Kuhad RC. Production of ganoderic acid by Ganoderma lucidum RCKB-2010 and its therapeutic potential. Ann. Microbiol. 64: 839–846 (2014).

    Article  CAS  Google Scholar 

  • Vu VH, Pham TA, Kim K. Improvement of a fungal strain by repeated and sequential mutagenesis and optimization of solid-state fermentation for the hyper-production of raw-starch-digesting enzyme. J. Microbiol. Biotechnol. 20: 718–726 (2010)

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Quan Y, Zhou F. Optimization of medium composition for exopolysacchride production by Phellinus nigricans. Carbohydr. Polym. 105: 200–206 (2014)

    Article  CAS  PubMed  Google Scholar 

  • Wei ZH, Duan YY, Qian YQ, Guo XF, Li YJ, Jin SH, Zhou ZX, Shan SY, Wang CR, Chen XJ, Zheng YG, Zhong JJ. Screening of Ganoderma strains with high polysaccharides and ganoderic acid contents and optimization of the fermentation medium by statistical methods. Bioprocess Biosyst. Eng. 37: 1789–1797 (2014)

    Article  CAS  PubMed  Google Scholar 

  • Yang FC, Ke YF, Kuo SS. Effect of fatty acids on the mycelial growth and polysaccharide formation by Ganoderma lucidum in shake flask cultures. Enzyme Microb. Technol. 27: 295–301 (2000)

    Article  CAS  PubMed  Google Scholar 

  • Yang FC, Liau CB. The influence of environmental conditions on polysaccharide formation by Ganoderma lucidum in submerged cultures. Process Biochem. 33: 547–553 (1998).

    Article  CAS  Google Scholar 

  • Yang FC, Yang MJ, Cheng SH. A novel method to enhance the mycelia production of Ganoderma lucidum in submerged cultures by polymer additives and agitation strategies. J. Taiwan Inst. Chem. Eng. 40: 148–154 (2009)

    Article  CAS  Google Scholar 

  • Yang SQ, Xiong H, Yang HY, Yan QJ, Jiang ZQ. High-level production of ß-1,3-1,4-glucanase by Rhizomucor miehei under solid-state fermentation and its potential application in the brewing industry. J. Appl. Microbiol. 118: 84–91 (2014)

    Article  CAS  PubMed  Google Scholar 

  • Yen HW, Chiang MH. Using the mycelium-covered cereals as an efficient inoculation method for rapamycin fermentation in a 15-L fermenter using Streptomyces hygroscopicus. Bioresour. Bioprocess 2: 43 (2015)

    Article  Google Scholar 

  • Zhao HZ, Wang J, Lu ZX. Optimization of process parameters of the Pholiota squarrosa extracellular polysaccharide by Box–Behnken statistical design. Carbohydr. Polym. 77: 677–680 (2009)

    Article  CAS  Google Scholar 

  • Zhu LW, Zhong JJ, Tang YJ. Significance of fungal elicitors on the production of ganoderic acid and Ganoderma polysaccharides by the submerged culture of medicinal mushroom Ganoderma lucidum. Process Biochem. 43: 1359–1370 (2008)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank the financial support by the Program for Young and Middle School Teachers of Education Department of Fujian Province (Grant No. JA15558), Ningde Normal University (Grant No. 2013F22), and the Industry Guidance Program of the Department of Science and Technology of Fujian Province (Grant No. 2015N0032).

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Liu, SR., Zhang, WR. Optimization of submerged culture conditions involving a developed fine powder solid seed for exopolysaccharide production by the medicinal mushroom Ganoderma lucidum. Food Sci Biotechnol 28, 1135–1145 (2019). https://doi.org/10.1007/s10068-018-0536-5

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