Ammonium Acetate Supplement Strategy for Enhancement of Chaetominine Production in Liquid Culture of Marine-Derived Aspergillus fumigatus CY018

  • Liu, Chang-Qing (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Wei, Xing-Chen (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • An, Fa-Liang (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Lu, Yan-Hua (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
  • Received : 2018.12.19
  • Accepted : 2019.02.01
  • Published : 2019.04.28


Pharmacological research on (CHA), a marine-derived quinazolinone alkaloid with significant cytotoxic activity, is restricted by low yields and is a problem that needs to be settled urgently. In this work, the selection of additional nitrogen sources and the optimization of additional concentrations and longer fermentation times using ammonium acetate, were investigated. CHA production was optimized to 62.1 mg/l with the addition of 50 mM ammonium acetate at 120 h of the fermentation in the shaker flask. This feeding strategy significantly increased 3-deoxy-arabino-heptulosonate-7-phosphate synthase activity and transcript levels of critical genes (laeA, dahp, and trpC) in the shikimate pathway compared with the non-treatment group. In addition, the selection of the feeding rate (0.01 and $0.03g/l{\cdot}h$) was investigated in a 5-L bioreactor. As a result, CHA production was increased by 57.9 mg/l with a $0.01g/l{\cdot}h$ ammonium acetate feeding rate. This work shows that the strategy of ammonium acetate supplementation had an effective role in improving CHA production by Aspergillus fumigatus CY018. It also shows that this strategy could serve as an important example of large-scale fermentation of a marine fungus in submerged culture.


Supported by : National Natural Science Foundation of China, Central Universities


  1. Sashidhara KV, White KN, Crews P. 2009. A Selective Account of effective paradigms and significant outcomes in the discovery of inspirational marine natural products. J. Nat. Prod. 72: 588-603.
  2. Liao L, Bae SY, Won TH, You M, Kim SH, Oh DC, et al. 2017. Asperphenins A and B, lipopeptidyl benzophenones from a marine-derived Aspergillus sp. Fungus. Org. Lett. 19: 2066-2069.
  3. Wang X, Yu H, Zhang Y, Lu X, Wang B, Liu X. 2018. Bioactive pimarane-type diterpenes from marine organisms. Chem. Biodivers. 15: e1700276.
  4. Duarte K, Rocha-Santos TAP, Freitas AC, Duarte AC. 2012. Analytical techniques for discovery of bioactive compounds from marine fungi. Trends Analyt. Chem. 34: 97-110.
  5. Lindequist U. 2016. Marine-derived pharmaceuticals - challenges and opportunities. Biomol. Ther. 24: 561-571.
  6. Zhang XY, Bao J, Zhong J, Xu XY, Nong XH, Qi SH. 2013. Enhanced production of a novel cytotoxic chromone oxalicumone A by marine-derived mutant Penicillium oxalicum SCSIO 24-2. Appl. Microbiol. Biotechnol. 97: 9657-9663.
  7. Zhang G, Li J, Zhu T, Gu Q, Li D. 2016. Advanced tools in marine natural drug discovery. Curr. Opin. Biotechnol. 42: 13-23.
  8. Agrawala S, Adholeya A, Barrow CJ, Deshmukh SK. 2018. Marine fungi: an untapped bioresource for future cosmeceuticals. Phytochem. Lett. 23: 15-20.
  9. Li XJ, Zhang Q, Zhang AL, Gao JM. 2012. Metabolites from Aspergillus fumigatus, an endophytic fungus associated with Melia azedarach, and their antifungal, antifeedant, and toxic activities. J. Agric. Food Chem. 60: 3424-3431.
  10. Li YX, Himaya SW, Dewapriya P, Zhang C, Kim SK. 2013. Fumigaclavine C from a marine-derived fungus Aspergillus Fumigatus induces apoptosis in MCF-7 breast cancer cells. Mar. Drugs 11: 5063-5086.
  11. Yao JY, Jiao RH, Liu CQ, Zhang YP, Yu WG, Lu YH, et al. 2016. Assessment of the cytotoxic and apoptotic effects of Chaetominine in a human leukemia cell line. Biomol. Ther. 24: 147-155.
  12. Yao JY, Wei X, Lu YH. 2016. Chaetominine reduces MRP1-mediated drug resistance via inhibiting PI3K/Akt/Nrf2 signaling pathway in K562/Adr human leukemia cells. Biochem. Biophys. Res. Commun. 473: 867-873.
  13. Stanley A, Punil Kumar HNP, Mutturi S, Vijayendra SVN. 2018. Fed-batch strategies for production of PHA using a native isolate of Halomonas venusta KT832796 strain. Appl. Biochem. Biotechnol. 184: 935-952.
  14. Wang DH, Chen FF, Wei GY, Jiang M, Dong MS. 2015. The mechanism of improved pullulan production by nitrogen limitation in batch culture of Aureobasidium pullulans. Carbohydr. Polym. 127: 325-331.
  15. Wang XYZ, Dong JJ, Xu GC, Han RZ, Ni Y. 2016. Enhanced curdlan production with nitrogen feeding during polysaccharide synthesis by Rhizobium radiobacter. Carbohydr. Polym. 150: 385-391.
  16. Liu CQ, Pan ZH, An FL, Lu YH. 2018. Co-addition strategy for enhancement of chaetominine from submerged fermentation of Aspergillus fumigatus CY018. Appl. Biochem. Biotechnol. 186: 384-399.
  17. Chludzinski AM, Salter DS, Nasser D. 1972. Feedback regulation of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthetase from a marine bacterium, Vibrio MB22. J. Bacteriol. 109: 1162-1169.
  18. Hu PS, Meng Y , Wise RP. 2009. Functional contribution of chorismate synthase, anthranilate synthase, and chorismate mutase to penetration resistance in Barley-Powdery mildew interactions. Mol. Plant Microbe. Interact. 22: 311-320.
  19. Culbertson JE, Chung DH, Ziebart KT, Espiritu E, Toney MD. 2015. Conversion of aminodeoxychorismate synthase into anthranilate synthase with janus mutations: mechanism of pyruvate elimination catalyzed by chorismate enzymes. Biochemistry 54: 2372-2384.
  20. Bok JW, Keller NP. 2004. LaeA, a regulator of secondary metabolism in Aspergillus spp. Eukaryot Cell 3: 527-535.
  21. Xie WP, Ye LD, Lv XM, Xu HM, Yu HW. 2015. Sequential control of biosynthetic path ways for balanced utilization of metabolic intermediates in Saccharomyces cerevisiae. Metab. Eng. 28: 8-18.
  22. Kong M, Wang FJ, Tian LY, Tang H, Zhang LP. 2018. Functional identification of glutamate cysteine ligase and glutathione synthetase in the marine yeast Rhodosporidium diobovatum. Naturwissenschaften 105: 4-12.
  23. Zhao W, Xu JW, Zhong JJ. 2011. Enhanced production of ganoderic acids in static liquid culture of Ganoderma lucidum under nitrogen-limiting conditions. Bioresour. Technol. 102: 8185-8190.
  24. Zhang YP, Jiao RH, Lu YH, Yao LY. 2016. Improvement of chaetominine production by tryptophan feeding and medium optimization in submerged fermentation of Aspergillus fumigatus CY018. Bioresour. Bioprocess. 3: 45-53.
  25. Zhu YX, Yao LY, Jiao RH, Lu YH, Tan RX. 2014. Enhanced production of Fumigaclavine C in liquid culture of Aspergillus fumigatus under a two-stage process. Bioresour. Technol. 152: 162-168.
  26. Liu CQ, Jiao RH, Yao LY, Zhang YP, Lu YH, Tan RX. 2016. Adsorption characteristics and preparative separation of chaetominine from Aspergillus fumigatus mycelia by macroporous resin. J. Chromatogr. B. 1015: 135-141.
  27. Li SB, Liu LM, Chen J. 2015. Mitochondrial fusion and fission are involved in stress tolerance of Candida glabrata. Bioresour. Bioprocess 2: 12-20.
  28. Lindlbauer KA, Marx H, Sauer M. 2017. Effect of carbon pulsing on the redox household of Lactobacillus diolivorans in order to enhance 1,3-propanediol production. N. Biotechnol. 34: 32-39.
  29. Yang SX, Chen XL, Xu N, Liu LM, Chen J. 2014. Urea enhances cell growth and pyruvate production in Torulopsis glabrata. Biotechnol. Prog. 30: 19-27.
  30. Lu YH, Pan ZH, Tao J, An FA. 2018. Induced effect of $Ca^{2+}$ on dalesconols A and B biosynthesis in the culture of Daldinia eschscholzii via calcium/calmodulin signaling. J. Biosci. Bioeng. 125: 205-210.
  31. Long CN, Cheng YJ, Cui JJ, Liu J, Gan LH, Zeng B, Long MN. 2018. Enhancing cellulase and hemicellulase production in Trichoderma orientalis EU7-22 via knockout of the creA. Mol. Biotechnol. 60: 55-61.
  32. Pandey SS, Singh S, Babu CSV, Shanker K, Srivastava NK, Shukla AK, et al. 2016. Fungal endophytes of Catharanthus roseus enhance vindoline content by modulating structural and regulatory genes related to terpenoid indole alkaloid biosynthesis. Sci. Rep. 6: 26583-26596.
  33. Zhang H, Hu YD, Lu RQ, Xia YJ, Zhang BB, Xu GR. 2014. Integrated strategy of pH-shift and glucose feeding for enhanced production of bioactive Antrodin C in submerged fermentation of Antrodia camphorate. J. Ind. Microbiol. Biotechnol. 41: 1305-1310.
  34. Roubos JA, Krabben P, de Laat WT, Babuska R, Heijnen JJ. 2002. Clavulanic acid degradation in Streptomyces clavuligerus fed-batch cultivations. Biotechnol. Prog. 18: 451-457.
  35. Mayer AF, Deckwer WD. 1996. Simultaneous production and decomposition of clavulanic acid during Streptomyces clavuligerus cultivations. Appl. Microbiol. Biotechnol. 45: 41-46.