Abstract
Huperzine-A (HupA) is an emerging, powerful, and promising natural acetylcholinesterase inhibitor. Despite that, the achieved yields of HupA from microbial sources are still far from the industrial applications. Accordingly, this paper was conducted to valorize solid-state fermentation (SSF) as an efficient production platform of HupA. Four agro-industrial wastes, namely rice bran, potato peel, sugarcane bagasse, and wheat bran, were tested and screened as cultural substrates for the production of HupA by the endophytic Alternaria brassica under SSF. Maximum HupA production was attained on using rice bran moistened by Czapex’s dox mineral broth. In the effort to increase the HupA titer, supplementation of the best moistening agent by different carbon and nitrogen sources was successfully investigated. Additionally, factors affecting HupA production under SSF including substrate concentration, moistening level, and inoculum concentration were optimized using response surface methodology. A Box-Behnken design was applied for generating a predictive model of the interactions between these factors. Under the optimum conditions of 15 g rice bran, inoculum concentration of 5 × 106 spores mL−1, and 60% moisture level, HupA concentration was intensified to 518.93 μg g−1. Besides, HupA production by the fungal strain was further enhanced using gamma-irradiation mutagenesis. The final HupA production was significantly intensified following exposure to 0.5 KGy gamma radiation to 1327 μg g−1, which represents a 12.85-fold increase. This is the first report on the successful production of the natural fungal metabolite HupA under SSF. Moreover, the achieved yield in this study using agro-industrial wastes may contribute to reducing the cost of HupA manufacture.
Key points
• Different agro-industrial by-products were tried as cultural substrates for the production of the acetylcholinesterase inhibitor HupA under SSF for the first time.
• Factors affecting HupA production under SSF were optimized using response surface methodology.
• The final HupA production was intensified following exposure to gamma radiation recording 1327 μg g −1 , which represents a 12.85-fold increase.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are included in this published article.
References
Ageitos AM, Garcia-Fuentes M (2019) Advances in drug delivery strategies for microbial healthcare products. In: Arora D, Sharma C, Jaglan S, Lichtfouse E (eds) Pharmaceuticals from microbes, Springer International Publishing, Springer Nature Switzerland. https://doi.org/10.1007/978-3-030-01881-8_1
Barrios-González J, Tarragó-Castellanos MR (2017) Solid-state fermentation: special physiology of fungi. In: Mérillon J-M, Ramawat KG (eds) Fungal metabolites. Springer International Publishing, Switzerland. Springer International Publishing, Switzerland. https://doi.org/10.1007/978-3-319-19456-1_6-1
Box GEP, Behnken DW (1960) Some new three level designs for the study of quantitative variables. Technometrics 2:455–475. https://doi.org/10.2307/1266454
Chiu SW, Chan SM (1992) Production of pigments by Monascuspurpurea using sugar-cane bagasse in roller bottle cultures. World J Microbiol Biotechnol 8:68–70. https://doi.org/10.1007/BF01200689
Delabona P, Pirota R, Codima C, Tremacoldi C, Rodrigues A, Farinas C (2012) Using Amazon forest fungi and agricultural residues as a strategy to produce cellulolytic enzymes. Biomass Bioenergy 37:243–250. https://doi.org/10.1016/j.biombioe.2011.12.006
El-Sayed ER (2021) Discovery of the anticancer drug vinblastine from the endophytic Alternariaalternata and yield improvement by gamma irradiation mutagenesis. J Appl Microbiol. https://doi.org/10.1111/jam.15169
El-Sayed ER, Ismaiel AA, Ahmed AS, Hassan IA, Karam El-Din AA (2019a) Bioprocess optimization using response surface methodology for production of the anticancer drug paclitaxel by Aspergillus fumigatus and Alternariatenuissima: enhanced production by ultraviolet and gamma irradiation. Biocatal Agric Biotechnol 18:100966. https://doi.org/10.1016/j.bcab.2019.01.034
El-Sayed ER, Ahmed AS, Ismaiel AA (2019b) Agro-industrial byproducts for production of the immunosuppressant mycophenolic acid by Penicilliumroqueforti under solid-state fermentation: enhanced production by ultraviolet and gamma irradiation. Biocatal Agric Biotechnol 18:101015. https://doi.org/10.1016/j.bcab.2019.01.053
El-Sayed ER, Ahmed AS, Al-Hagar OEA (2020a) Agro-industrial wastes for production of paclitaxel by irradiated Aspergillus fumigatus under solid-state fermentation. J Appl Microbiol 128:1427–1439. https://doi.org/10.1111/jam.14574
El-Sayed ER, Abdelhakim HK, Ahmed AS (2020b) Solid-state fermentation for enhanced production of selenium nanoparticles by gamma-irradiated Monascuspurpureus and their biological evaluation and photocatalytic activities. Bioproc Biosyst Eng 43:797–809. https://doi.org/10.1007/s00449-019-02275-7
El-Sayed ER, Zaki AG, Ahmed AS, Ismaiel AA (2020c) Production of the anticancer drug taxol by the endophytic fungus Epicoccumnigrum TXB502: enhanced production by gamma irradiation mutagenesis and immobilization technique. Appl Microbiol Biotechnol 104:6991–7003. https://doi.org/10.1007/s00253-020-10712-x
El-Sayed ER, Abdelhakim HK, Zakaria Z (2020d) Extracellular biosynthesis of cobalt ferrite nanoparticles by Monascuspurpureus and their antioxidant, anticancer and antimicrobial activities: yield enhancement by gamma irradiation. Mater Sci Eng C 107:110318. https://doi.org/10.1016/j.msec.2019.110318
El-Sayed ER, Ahmed AS, Abdelhakim HK (2020e) A novel source of the cardiac glycoside digoxin from the endophytic fungus Epicoccumnigrum: isolation, characterization, production enhancement by gamma irradiation mutagenesis and anticancer activity evaluation. J Appl Microbiol 128:747–762. https://doi.org/10.1011/JAM.14510
El-Sayed ER, Ahmed AS, Hassan IA, Ismaiel AA, Zahraa A, El AK (2020f) Semi-continuous production of the anticancer drug taxol by Aspergillus fumigatus and Alternariatenuissima immobilized in calcium alginate beads. Bioprocess Biosyst Eng 43:997–1008. https://doi.org/10.1007/s00449-020-02295-8
Fiedurek J, Trytek M, Szczodrak J (2017) Strain improvement of industrially important microorganisms based on resistance to toxic metabolites and abiotic stress. J Basic Microbiol 57:445–459. https://doi.org/10.1002/jobm.201600710
Kalogeris E, Christakopoulos P, Katapodis P, Alexiou A, Vlachou S, Kekos D, Macris BJ (2003) Production and characterization of cellulolytic enzymes from the thermophilic fungus Thermoascusaurantiacus under solid state cultivation of agricultural wastes. Process Biochem 38:1099–1104. https://doi.org/10.1016/S0032-9592(02)00242-X
Kalpanadevi C, Singh V, Subramanian R (2018) Influence of milling on the nutritional composition of bran from different rice varieties. J Food Sci Technol 55:2259–2269. https://doi.org/10.1007/s13197-018-3143-9
Kashyap P, Sabu A, Pandey A, Szakacs S, Soccol CR (2002) Extra-cellular L-glutaminase production by Zygosaccharomycesrouxii under solid-state fermentation. Process Biochem 38:307–312. https://doi.org/10.1016/S0032-9592(02)00060-2
Kennedy M, Krouse D (1999) Strategies for improving fermentation medium performance: a review. J Ind Microbiol Biotechnol 23:456–475. https://doi.org/10.1038/sj.jim.2900755
Khir R, Pan Z (2019) Chapter 2: Rice. In: Pan Z, Zhang R, Zicari S (eds) Integrated processing technologies for food and agricultural by-products. Elsevier Academic Press, London, UK. https://doi.org/10.1016/B978-0-12-814138-0.00002-2
Koshiba T, Yokoshima S, Fukuyama T (2014) Correction to total synthesis of (−)-huperzine A. Org Lett 16:1270–1270. https://doi.org/10.1021/acs.orglett.7b03555
Krishania M, Sindhu R, Binod P, Ahluwalia V, Kumar V, Sangwan RS, Pandey A (2015) Chapter 5 - design of bioreactors in solid-state fermentation. In: Pandey A, Larroche C, Socco C, (eds) Current developments in biotechnology and bioengineering. Elsevier. https://doi.org/10.1016/B978-0-444-63990-5.00005-0
Ishiuchi K, Park JJ, Long RM, Gang DR (2013) Production of huperzine A and other lycopodium alkaloids in Huperzia species grown under controlled conditions and in vitro. Phytochemistry 91:208–219. https://doi.org/10.1016/j.phytochem.2012.11.012
Ma X, Gang DR (2008) In vitro production of huperzine A, a promising drug candidate for Alzheimer’s disease. Phytochemistry 69:2022–2028. https://doi.org/10.1016/j.phytochem.2008.04.017
Manan MA, Webb C (2017) Design aspects of solid state fermentation as applied to microbial bioprocessing. J Appl Biotechnol Bioeng 4:511–532. https://doi.org/10.15406/jabb.2017.04.00094
Mousa SA, El-Sayed ER, Mohamed SS, Abo El-Seoud MA, Elmehlawy AA, Abdou DAM (2021) Novel mycosynthesis of Co3O4, CuO, Fe3O4, NiO, and ZnO nanoparticles by the endophytic Aspergillus terreus and evaluation of their antioxidant and antimicrobial activities. Appl Microbiol Biotechnol 105:741–753. 10. 1007/s00253–020–11046–4
Ramana Murthy MV, Mohan EVS, SadhuKhan AK (1999) Cyclosporin-A production by Tolypocladiuminflatum using solid state fermentation. Process Biochem 34:269–280. https://doi.org/10.1016/S0032-9592(98)00095-8
Schmidt CG, Goncalves LM, Prietto L, Hackbart HS, Furlong EB (2014) Antioxidant activity and enzyme inhibition of phenolic acids from fermented rice bran with fungus Rizhopusoryzae. Food Chem 146:371–377. https://doi.org/10.1016/j.foodchem.2013.09.101
Pandey A (2003) Solid-state fermentation. Biochem Eng J 13:81–84. https://doi.org/10.1016/S1369-703X(02)00121-3
Parekh S, Vinci VA, Strobel RJ (2000) Improvement of microbial strains and fermentation processes. Appl Microbiol Biotechnol 54:287–301. https://doi.org/10.1007/s002530000403
Paster N, Barkai-Golan R, Padova R (1985) Effect of gamma radiation on ochratoxin production by the fungus Aspergillus ochraceus. J Sci Food Agric 36:445–449. https://doi.org/10.1002/jsfa.2740360604
Sakurai Y, Lee TH, Shiota H (1977) On the convenient method for glucosamine estimation in koji. Agric Biol Chem 44:619–624. https://doi.org/10.1080/00021369.1977.10862552
Sharma A, Vivekanand V, Singh RP (2018) Solid-state fermentation for gluconic acid production from sugarcane molasses by Aspergillus niger ARNU-4 employing tea waste as the novel solid support. Biores Technol 99:3444–3450. https://doi.org/10.1016/j.biortech.2007.08.006
Shu S, Zhao X, Wang W, Zhang G, Cosoveanu A, Ahn Y, Wang M (2014) Identification of a novel endophytic fungus from Huperziaserrata which produces huperzine A. World J Microbiol Biotechnol 30:3101–3109. https://doi.org/10.1007/s11274-014-1737-6
Thacker J (1999) Repair of ionizing radiation damage in mammalian cells. Alternative pathways and their fidelity. CR Acad Sci III 322:103–108. https://doi.org/10.1016/S0764-4469(99)80030-4
Viniegra-Gonzalez G, Favela-Torres E (2006) Why solid-state fermentation seems to be resistant to catabolite repression? Food Technol Biotechnol 44:397–406
Wang R, Yan H, Tang XC (2006) Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. Acta Pharmacol Sin 27:1–26. https://doi.org/10.1111/j.1745-7254.2006.00255.x
Wang Y, Zeng QG, Zhang ZB, Yan RM, Wang LY, Zhu D (2011) Isolation and characterization of endophytic huperzine A-producing fungi from Huperziaserrata. J Ind Microbiol Biotechnol 38:1267–1278. https://doi.org/10.1007/s10295-010-0905-4
Yang G, Wang Y, Tian J, Liu P (2013) Huperzine A for Alzheimer’s disease: a systematic review and meta-analysis of randomized clinical trials. PLoS on 23(8):e74916. https://doi.org/10.1371/journal.pone.0074916
Zaki AG, El-Sayed ER, AbdElkodous M, El-Sayyad GS (2020) Microbial acetylcholinesterase inhibitors for Alzheimer’s therapy: recent trends on extraction, detection, irradiation-assisted production improvement and nano-structured drug delivery. Appl Microbiol Biotechnol 104:4717–4735. https://doi.org/10.1007/s00253-020-10560-9
Zaki AG, El-Shatoury EH, Ahmed AS, Al-Hagar OEA (2019) Production and enhancement of the acetylcholinesterase inhibitor, huperzine A, from an endophytic Alternariabrassicae AGF041. Appl Microbiol Biotechnol 103:5867–5878. https://doi.org/10.1007/s00253-019-09897-7
Zaki AG, El-Shatoury EH, Ahmed AS, Al-Hagar OEA (2021) Response surface methodology-mediated improvement of the irradiated endophytic fungal strain, Alternariabrassicae AGF041 for Huperzine A-hyperproduction. Lett Appl Microbiol 72:427–437. https://doi.org/10.1111/lam.13435
Zhu D, Wang J, Zeng Q, Zhang Z, Yan R (2010) A novel endophytic Huperzine A-producing fungus, Shiraia sp. Slf14, isolated from Huperziaserrata. J Appl Microbiol 109:1469–1478. https://doi.org/10.1111/j.1365-2672.2010.04777.x
Author information
Authors and Affiliations
Contributions
A. G. Z. suggested the research point, designed the experimental methodology, conducted experiments, analyzed the obtained data, and wrote the whole manuscript. E. R. E. suggested the research point, designed the experimental methodology, conducted the experiments, analyzed the obtained data, and wrote the whole manuscript.
Corresponding author
Ethics declarations
Ethical approval
This article does not contain any studies with human participants or animals performed by the author.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Zaki, A.G., El-Sayed, ES.R. New and potent production platform of the acetylcholinesterase inhibitor huperzine A by gamma-irradiated Alternaria brassicae under solid-state fermentation. Appl Microbiol Biotechnol 105, 8869–8880 (2021). https://doi.org/10.1007/s00253-021-11678-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-021-11678-0