Abstract
In this study, endophytic fungi were isolated from Dioscorea zingiberensis C.H. Wright (DZW), and a novel clean process to prepare diosgenin from DZW was developed. A total of 123 strains of endophytic fungi were isolated from different plant tissues of DZW. Among them, the strain Fusarium sp. (CPCC 400709) showed the best activity of hydrolyzing steroidal saponins in DZW into diosgenin. Thus, this strain was used to prepare diosgenin from DZW by solid-state fermentation. The fermentation parameters were optimized using response surface methodology, and a high yield of diosgenin (2.16%) was obtained at 14.5% ammonium sulfate, an inoculum size of 12.3%, and 22 days of fermentation. Furthermore, the highest diosgenin yield (2.79%) was obtained by co-fermentation with Fusarium sp. (CPCC 400709) and Curvularia lunata (CPCC 400737), which was 98.9% of that obtained by β-glucosidase pretreated acid hydrolysis (2.82%). This process is acid-free and wastewater-free, and shows promise as an effective and clean way to prepare diosgenin for use in industrial applications from DZW.
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References
Abildgren MP, Lund F, Thrane U, Elmholt S (1987) Czapek-Dox agar containing iprodione and dicloran as a selective medium for the isolation of Fusarium species. Lett Appl Microbiol 5(4):83–86. https://doi.org/10.1111/j.1472-765X.1987.tb01620.x
Azin M, Moravej R, Zareh D (2007) Production of xylanase by Trichoderma longibrachiatum on a mixture of wheat bran and wheat straw: optimization of culture condition by Taguchi method. Enzym Microb Technol 40(4):801–805. https://doi.org/10.1016/j.enzmictec.2006.06.013
Bai Y, Zhang LW, Jin WW, Wei M, Zhou PP, Zheng GH, Niu LL, Nie L, Zhang YL, Wang HY, Yu LJ (2015) In situ high-valued utilization and transformation of sugars from Dioscorea zingiberensis C.H. Wright for clean production of diosgenin. Bioresour Technol 196:642–647. https://doi.org/10.1016/j.biortech.2015.08.010
Bier MCJ, Medeiros ABP, Soccol CR (2017) Biotransformation of limonene by an endophytic fungus using synthetic and orange residue-based media. Fungal Biol 121(2):137–144. https://doi.org/10.1016/j.funbio.2016.11.003
Čertík M, Adamechová Z, Guothová L (2013) Simultaneous enrichment of cereals with polyunsaturated fatty acids and pigments by fungal solid state fermentations. J Biotechnol 168(2):130–134. https://doi.org/10.1016/j.jbiotec.2013.03.016
Chen HZ (2013) Modern solid state fermentation-theory and practice. Springer, Dordrecht
Chen HZ, He Q (2012) Value-added bioconversion of biomass by solid-state fermentation. J Chem Technol Biotechnol 87(12):1619–1625. https://doi.org/10.1002/jctb.3901
Cheng P, Zhao HZ, Zhao B, Ni JR (2009) Pilot treatment of wastewater from Dioscorea zingiberensis C.H. Wright production by anaerobic digestion combined with a biological aerated filter. Bioresour Technol 100(12):2918–2925. https://doi.org/10.1016/j.biortech.2009.01.054
Cheng YT, Dong C, Huang CC, Zhu YL (2015) Enhanced production of diosgenin from Dioscorea zingiberensis in mixed culture solid state fermentation with Trichoderma reesei and Aspergillus fumigatus. Biotechnol Biotechnol Equip 29(4):773–778. https://doi.org/10.1080/13102818.2015.1032352
Cubero OF, Crespo A, Fatehi J, Bridge PD (1999) DNA extraction and PCR amplification method suitable for fresh, herbarium-stored, lichenized, and other fungi. Plant Syst Evol 216(3–4):243–249. https://doi.org/10.1007/Bf01084401
Dean R, Van Kan JAL, Pretorius ZA, Hammond-Kosack KE, Di Pietro A, Spanu PD, Rudd JJ, Dickman M, Kahmann R, Ellis J, Foster GD (2012) The top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 13(4):414–430. https://doi.org/10.1111/j.1364-3703.2011.00783.x
Doberski JW, Tribe HT (1980) Isolation of entomogenous fungi from elm bark and soil with reference to ecology of Beauveria bassiana and Metarhizium anisopliae. Trans Br Mycol Soc 74(Feb):95–100. https://doi.org/10.1016/S0007-1536(80)80013-1
Dong AL, Ye M, Guo HZ, Zheng JH, Guo D (2003) Microbial transformation of ginsenoside Rb1 by Rhizopus stolonifer and Curvularia lunata. Biotechnol Lett 25(4):339–344. https://doi.org/10.1023/a:1022320824000
Dong YS, Teng H, Qi SS, Liu L, Wang H, Zhao YK, Xiu ZL (2010) Pathways and kinetics analysis of biotransformation of Dioscorea zingiberensis by Aspergillus oryzae. Biochem Eng J 52(2–3):123–130. https://doi.org/10.1016/j.bej.2010.07.011
Dong JZ, Lei C, Lu DY, Wang Y (2015) Direct biotransformation of dioscin into diosgenin in rhizome of Dioscorea zingiberensis by Penicillium dioscin. Indian J Microbiol 55(2):200–206. https://doi.org/10.1007/s12088-014-0507-3
Fang H, Zhao C, Song XY (2010) Optimization of enzymatic hydrolysis of steam-exploded corn stover by two approaches: response surface methodology or using cellulase from mixed cultures of Trichoderma reesei RUT-C30 and Aspergillus niger NL02. Bioresour Technol 101(11):4111–4119. https://doi.org/10.1016/j.biortech.2010.01.078
Feng B, Hu W, Ma BP, Wang YZ, Huang HZ, Wang SQ, Qian XH (2007) Purification, characterization, and substrate specificity of a glucoamylase with steroidal saponin-rhamnosidase activity from Curvularia lunata. Appl Microbiol Biotechnol 76(6):1329–1338. https://doi.org/10.1007/s00253-007-1117-3
Feng B, Huang HZ, Zhou WB, Kang LP, Zou P, Liu YX, Yu HS, Han BQ, Li YY, Zhang LL, Zhang T, Ma BP (2010) Substrate specificity, purification and identification of a novel pectinase with the specificity of hydrolyzing the α-1,4-glycosyl residue in steroidal saponin. Process Biochem 45(8):1383–1392. https://doi.org/10.1016/j.procbio.2010.05.009
Fu YY, Yu HS, Tang SH, Hu XC, Wang YH, Liu B, Yu CX, Jin FX (2010) New dioscin-glycosidase hydrolyzing multi-glycosides of dioscin from Absidia strain. J Microbiol Biotechnol 20(6):1011–1017. https://doi.org/10.4014/jmb.0910.10039
Garai D, Kumar V (2013) A Box–Behnken design approach for the production of xylanase by Aspergillus candidus under solid state fermentation and its application in saccharification of agro residues and Parthenium hysterophorus L. Ind Crop Prod 44:352–363. https://doi.org/10.1016/j.indcrop.2012.10.027
He ZM, Chen HY, Li GF, Zhu HY, Gao YG, Zhang LX, Sun JM (2014) Diosgenin inhibits the migration of human breast cancer MDA-MB-231 cells by suppressing Vav2 activity. Phytomedicine 21(6):871–876. https://doi.org/10.1016/j.phymed.2014.02.002
Hölker U, Lenz J (2005) Solid-state fermentation—are there any biotechnological advantages? Curr Opin Microbiol 8(3):301–306. https://doi.org/10.1016/j.mib.2005.04.006
Huang W, Zhao HZ, Ni JR, Zuo H, Qiu LL, Li H, Li H (2008) The best utilization of D. zingiberensis C.H. Wright by an eco-friendly process. Bioresour Technol 99(15):7407–7411. https://doi.org/10.1016/j.biortech.2008.01.015
Huang HZ, Zhao M, Lu L, Tan DW, Zhou WB, Xiong CQ, Zhao Y, Song XB, Yu LY, Ma BP (2013) Pathways of biotransformation of zingiberen newsaponin from Dioscorea zingiberensis C. H. Wright to diosgenin. J Mol Catal B Enzym 98:1–7. https://doi.org/10.1016/j.molcatb.2013.09.013
Jin ZX (2011) Study on the endophytes from Dioscorea zingiberensis C.H. Wright and metabolites of 2 isolates. Dissertation, Southwest University (in Chinese)
Kalailingam P, Kannaian B, Tamilmani E, Kaliaperumal R (2014) Efficacy of natural diosgenin on cardiovascular risk, insulin secretion, and beta cells in streptozotocin (STZ)-induced diabetic rats. Phytomedicine 21(10):1154–1161. https://doi.org/10.1016/j.phymed.2014.04.005
Kanmani P, Karthik S, Aravind J, Kumaresan K (2013) The use of response surface methodology as a statistical tool for media optimization in lipase production from the dairy effluent isolate Fusarium solani. ISRN Biotechnol 2013:8. https://doi.org/10.5402/2013/528708
Kollerov VV, Shutov AA, Fokina VV, Sukhodol’skaya GV, Gulevskaya SA, Donova MV (2010) Bioconversion of C19- and C21-steroids with parent and mutant strains of Curvularia lunata. Appl Biochem Microbiol 46(2):198–205. https://doi.org/10.1134/s0003683810020122
Kongklom N, Luo HZ, Shi ZP, Pechyen C, Chisti Y, Sirisansaneeyakul S (2015) Production of poly-γ-glutamic acid by glutamic acid-independent Bacillus licheniformis TISTR 1010 using different feeding strategies. Biochem Eng J 100:67–75. https://doi.org/10.1016/j.bej.2015.04.007
Krishna C (2005) Solid-state fermentation systems—an overview. Crit Rev Biotechnol 25(1–2):1–30. https://doi.org/10.1080/07388550590925383
Lei J, Niu H, Li TH, Huang W (2012) A novel beta-glucosidase from Aspergillus fumigates releases diosgenin from spirostanosides of Dioscorea zingiberensis C. H. Wright (DZW). World J Microbiol Biotechnol 28(3):1309–1314. https://doi.org/10.1007/s11274-011-0907-z
Liu L, Dong YS, Qi SS, Wang H, Xiu ZL (2010) Biotransformation of steriodal saponins in Dioscorea zingiberensis C. H. Wright to diosgenin by Trichoderma harzianum. Appl Microbiol Biotechnol 85(4):933–940. https://doi.org/10.1007/s00253-009-2098-1
Lonsane BK, Saucedo-Castaneda G, Raimbault M, Roussos S, Viniegra-Gonzalez G, Ghildyal NP, Ramakrishna M, Krishnaiah MM (1992) Scale-up strategies for solid state fermentation systems. Process Biochem 27(5):259–273. https://doi.org/10.1016/0032-9592(92)85011-P
Lv YC, Yang J, Yao F, Xie W, Tang YY, Ouyang XP, He PP, Tan YL, Li L, Zhang M, Liu D, Cayabyab FS, Zheng XL, Tang CK (2015) Diosgenin inhibits atherosclerosis via suppressing the MiR-19b-induced downregulation of ATP-binding cassette transporter A1. Atherosclerosis 240(1):80–89. https://doi.org/10.1016/j.atherosclerosis.2015.02.044
Pang X, Huang HZ, Zhao Y, Xiong CQ, Yu LY, Ma BP (2015) Conversion of furostanol saponins into spirostanol saponins improves the yield of diosgenin from Dioscorea zingiberensis by acid hydrolysis. RSC Adv 5(7):4831–4837. https://doi.org/10.1039/c4ra12709a
Percival Zhang YH, Himmel ME, Mielenz JR (2006) Outlook for cellulase improvement: screening and selection strategies. Biotechnol Adv 24(5):452–481. https://doi.org/10.1016/j.biotechadv.2006.03.003
Qi SS, Dong YS, Zhao YK, Xiu ZL (2009) Qualitative and quantitative analysis of microbial transformation of steroidal saponins in Dioscorea zingiberensis. Chromatographia 69(9–10):865–870. https://doi.org/10.1365/s10337-009-0978-2
Senthilkumar SR, Ashokkumar B, Chandra Raj K, Gunasekaran P (2005) Optimization of medium composition for alkali-stable xylanase production by Aspergillus fischeri Fxn 1 in solid-state fermentation using central composite rotary design. Bioresour Technol 96(12):1380–1386. https://doi.org/10.1016/j.biortech.2004.11.005
Sindhu I, Chhibber S, Capalash N, Sharma P (2006) Production of cellulase-free xylanase from Bacillus megaterium by solid state fermentation for biobleaching of pulp. Curr Microbiol 53(2):167–172. https://doi.org/10.1007/s00284-006-0051-4
Singhania RR, Patel AK, Soccol CR, Pandey A (2009) Recent advances in solid-state fermentation. Biochem Eng J 44(1):13–18. https://doi.org/10.1016/j.bej.2008.10.019
Soccol CR, da Costa ESF, Letti LAJ, Karp SG, Woiciechowski AL, de Souza Vandenberghe LP (2017) Recent developments and innovations in solid state fermentation. Biotechnol Res Innov 1(1):52–71. https://doi.org/10.1016/j.biori.2017.01.002
Strobel GA (2003) Endophytes as sources of bioactive products. Microb Infect 5(6):535–544. https://doi.org/10.1016/S1286-4579(03)00073-X
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(10):2731–2739. https://doi.org/10.1093/molbev/msr121
Wang Y, Wang C, Hang Y, Li G, Qiao Y, Wang Y, He M, Hua W (2015) Progress in the study of endophytes from Dioscorea zingiberensis. Genom Appl Biol 34(8):1808–1811
Wei M, Bai Y, Ao MZ, Jin WW, Yu PP, Zhu M, Yu LJ (2013) Novel method utilizing microbial treatment for cleaner production of diosgenin from Dioscorea zingiberensis C.H. Wright (DZW). Bioresour Technol 146:549–555. https://doi.org/10.1016/j.biortech.2013.07.090
Xu L, Zhou L, Zhao J, Li J, Li X, Wang J (2008) Fungal endophytes from Dioscorea zingiberensis rhizomes and their antibacterial activity. Lett Appl Microbiol 46(1):68–72. https://doi.org/10.1111/j.1472-765X.2007.02264.x
Yaderets VV, Andryushina VA, Bartoshevich YE, Domracheva AG, Novak MI, Stytsenko TS, Voishvillo NE (2007) A study of steroid hydroxylation activity of Curvularia lunata mycelium. Appl Biochem Microbiol 43(6):620–624. https://doi.org/10.1134/s0003683807060099
Yang SS, Guo WQ, Zhou XJ, Meng ZH, Liu B, Ren NQ (2011) Optimization of operating parameters for sludge process reduction under alternating aerobic/oxygen-limited conditions by response surface methodology. Bioresour Technol 102(21):9843–9851. https://doi.org/10.1016/j.biortech.2011.07.079
Yong XY, Raza W, Yu GH, Ran W, Shen QR, Yang XM (2011) Optimization of the production of poly-γ-glutamic acid by Bacillus amyloliquefaciens C1 in solid-state fermentation using dairy manure compost and monosodium glutamate production residues as basic substrates. Bioresour Technol 102(16):7548–7554. https://doi.org/10.1016/j.biortech.2011.05.057
Zhang JJ, Li H, Li H, Lu ZM, Shi JS, Xu ZL (2013a) Screening and condition optimization of a strain for efficiently biotransformation of saponins in Dioscorea zingiberensis into diosgenin. Chin J Biotechnol 29(6):848–852
Zhang T, Xiang HB, Zhang YQ, Liu HY, Wei YZ, Zhao LX, Yu LY (2013b) Molecular analysis of fungal diversity associated with three bryophyte species in the Fildes Region, King George Island, maritime Antarctica. Extremophiles 17(5):757–765. https://doi.org/10.1007/s00792-013-0558-0
Zhao YT, Xu ZL, Dai CC, Tang XL, Xia B, Wang Q (2007) Selection of the microorganism for dioscin hydrolyze. J Chin Med Mater 30(8):905–909
Zhu YL, Huang W, Ni JR (2010a) A promising clean process for production of diosgenin from Dioscorea zingiberensis C. H Wright. J Clean Prod 18(3):242–247. https://doi.org/10.1016/j.jclepro.2009.10.012
Zhu YL, Huang W, Ni JR, Liu W, Li H (2010b) Production of diosgenin from Dioscorea zingiberensis tubers through enzymatic saccharification and microbial transformation. Appl Microbiol Biotechnol 85(5):1409–1416. https://doi.org/10.1007/s00253-009-2200-8
Zhu YL, Ni JR, Huang W (2010c) Process optimization for the production of diosgenin with Trichoderma reesei. Bioprocess Biosyst Eng 33(5):647–655. https://doi.org/10.1007/s00449-009-0390-1
Funding
This work was supported by the National Science and Technology Project of China (Nos. 2018ZX09711001-007-001 and 2017ZX09301072), CAMS Innovation Fund for Medical Sciences (2016-I2M-2-002), the National Infrastructure of Microbial Resources (No.NIMR-2018-3), the National Natural Science Foundation of China (Nos. 81373452 and 81621064), and Li-yan Yu was supported by Xiehe Scholar.
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Xiang, H., Zhang, T., Pang, X. et al. Isolation of endophytic fungi from Dioscorea zingiberensis C. H. Wright and application for diosgenin production by solid-state fermentation. Appl Microbiol Biotechnol 102, 5519–5532 (2018). https://doi.org/10.1007/s00253-018-9030-5
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DOI: https://doi.org/10.1007/s00253-018-9030-5