Log in

Analysis of the L-malate biosynthesis pathway involved in poly(β-L-malic acid) production in Aureobasidium melanogenum GXZ-6 by addition of metabolic intermediates and inhibitors

  • Microbial Physiology and Biochemistry
  • Published:
Journal of Microbiology Aims and scope Submit manuscript

Abstract

Poly(β-L-malic acid) (PMA) is a promising polyester formed from L-malate in microbial cells. Malate biosynthesis is crucial for PMA production. Previous studies have shown that the non-oxidative pathway or oxidative pathway (TCA cycle) is the main biosynthetic pathway of malate in most of PMA-producing strains, while the glyoxylate cycle is only a supplementary pathway. In this study, we investigated the effect of exogenous metabolic intermediates and inhibitors of the malate biosynthetic pathway on PMA production by Aureobasidium melanogenum GXZ-6. The results showed that PMA production was stimulated by maleic acid (a fumarase inhibitor) and sodium malonate (a succinate dehydrogenase inhibitor) but inhibited by succinic acid and fumaric acid. This indicated that the TCA cycle might not be the only biosynthetic pathway of malate. In addition, the PMA titer increased by 18.1% upon the addition of glyoxylic acid after 72 h of fermentation, but the PMA titer decreased by 7.5% when itaconic acid (an isocitrate lyase inhibitor) was added, which indicated that malate for PMA production was synthesized significantly via the glyoxylate cycle rather than the TCA cycle. Furthermore, in vitro enzyme activities of the TCA and glyoxylate cycles suggested that the glyoxylate cycle significantly contributed to the PMA production, which is contradictory to what has been reported previously in other PMA-producing A. pullulans.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bott, M. and Eikmanns, B.J. 2013. TCA cycle and glyoxylate shunt of Corynebacterium glutamicum, pp. 281–313. In Hideaki, Y. and Masayuki, I. (eds.), Corynebacterium glutamicum, Springer, Berlin, Germany.

    Chapter  Google Scholar 

  • Cao, W., Luo, J., Qi, B., Zhao, J., Qiao, C., Ding, L., Su, Y., and Wan, Y. 2014. β-poly(L-malic acid) production by fed-batch culture of Aureobasidium pullulans ipe-1 with mixed sugars. Eng. Life Sci. 14, 180–189.

    Article  CAS  Google Scholar 

  • Chi, Z., Liu, G.L., Liu, C.G., and Chi, Z.M. 2016a. Poly(β-L-malic acid) (PMLA) from Aureobasidium spp. and its current proceedings. Appl. Microbiol. Biotechnol. 100, 3841–3851.

    Article  CAS  PubMed  Google Scholar 

  • Chi, Z., Wang, Z.P., Wang, G.Y., Khan, I., and Chi, Z.M. 2016b. Microbial biosynthesis and secretion of L-malic acid and its applications. Crit. Rev. Biotechnol. 36, 99–107.

    Article  CAS  PubMed  Google Scholar 

  • Chinnici, F., Spinabelli, U., Riponi, C., and Amati, A. 2005. Optimization of the determination of organic acids and sugars in fruit juices by ion-exclusion liquid chromatography. J. Food Compos. Anal. 18, 121–130.

    Article  CAS  Google Scholar 

  • Ding, H., Helguera, G., Rodriguez, J.A., Markman, J., Luria-Perez, R., Gangalum, P., Portilla-Arias, J., Inoue, S., Daniels-Wells, T.R., Black, K., et al. 2013. Polymalic acid nanobioconjugate for simultaneous immunostimulation and inhibition of tumor growth in HER2/neu-positive breast cancer. J. Control. Release 171, 322–329.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fischer, H., Erdmann, S., and Holler, E. 1989. An unusual polyanion from Physarum polycephalum that inhibits homologous DNApolymerasea in vitro. Biochemistry 28, 5219–5226.

    Article  CAS  PubMed  Google Scholar 

  • Garraway, M.O. and Evans, R.C. 1984. Fungal nutrition and physiology, pp. 298–312. Wiley, New York, USA.

    Google Scholar 

  • Giachetti, E., Pinzauti, G., Bonaccorsi, R., and Vanni, P. 1988. Isocitrate lyase from Pinus pinea. Characterization of its true substrate and the action of magnesium ions. Eur. J. Biochem. 172, 85–91.

    CAS  PubMed  Google Scholar 

  • Holler, E., Angerer, B., Achhammer, G., Miller, S., and Windisch, C. 1992. Biological and biosynthetic properties of poly-L-malate. FEMS Microbiol. Rev. 103, 109–118.

    CAS  Google Scholar 

  • Jong-Gubbels, P.D., Vanrolleghem, P., Heijnen, S., Dijken, J.P.V., and Pronk, J.T. 1995. Regulation of carbon metabolism in chemostat cultures of Saccharomyces cerevisiae grown on mixtures of glucose and ethanol. Yeast 11, 407–418.

    Article  PubMed  Google Scholar 

  • Lee, B.S. and Holler, E. 2000. β-poly(L-malate) production by nongrowing microplasmodia of Physarum polycephalum. Effects of metabolic intermediates and inhibitors. FEMS Microbiol. Lett. 193, 69–74.

    CAS  PubMed  Google Scholar 

  • Liu, S.J. and Steinbüchel, A. 1997. Production of poly(malic acid) from different carbon sources and its regulation in Aureobasidium pullulans. Biotechnol. Lett. 19, 11–14.

    Article  Google Scholar 

  • Livak, K.J. and Schmittgen, T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25, 402–408.

    Article  CAS  PubMed  Google Scholar 

  • Massey, V. 1953. Studies on fumarase. II. The effects of inorganic anions on fumarase activity. Biochem. J. 53, 67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nagata, N., Nakahara, T., and Tabuchi, T. 1993. Fermentative production of poly(β-L-malic acid), a polyelectrolytic biopolyester, by Aureobasidium sp. Biosci. Biotech. Biochem. 57, 638–642.

    Article  CAS  Google Scholar 

  • Rikhvanov, E.G., Varakina, N.N., Rusaleva, T.M., Rachenko, E.I., and Voinikov, V.K. 2003. The effect of sodium malonate on yeast thermotolerance. Microbiology 72, 548–552.

    Article  CAS  Google Scholar 

  • Schmidt, A., Windisch, C., and Holler, E. 1996. Nuclear accumulation and homeostasis of the unusual polymer β-poly(L-malate) in plasmodia of Physarum polycephalum. Eur. J. Cell Biol. 70, 373–380.

    CAS  PubMed  Google Scholar 

  • Shimada, K., Matsushima, K.I., Fukumoto, J., and Yamamoto, T. 1969. Poly-(L)-malic acid; A new protease inhibitor from Penicillium cyclopium. Biochem. Biophys. Res. Commun. 35, 619–624.

    Article  CAS  PubMed  Google Scholar 

  • Wallace, J.C., Jitrapakdee, S., and Chapmansmith, A. 1998. Pyruvate carboxylase. Int. J. Biochem. Cell 30, 1–5.

    Article  CAS  Google Scholar 

  • Yang, E.S., Richter, C., Chun, J.S., Huh, T.L., Kang, S.S., and Park, J.W. 2002. Inactivation of NADP+-dependent isocitrate dehydrogenase by nitric oxide. Free Radic. Biol. Med. 33, 927–937.

    Article  CAS  PubMed  Google Scholar 

  • Yang, X.T., Guo, Y.M., Zeng, X.F., Man, Y., Zheng, P., Wang, C.L., and Sun, J.B. 2011. Characterization of the hyperproduction process of citric acid by Aspergillus niger and the formation of glyoxylic acid. Mod. Food Sci. Technol. 27, 1183–1186.

    CAS  Google Scholar 

  • Zeng, W., Zhang, B., Chen, G.G., Li, M.X., and Liang, Z.Q. 2018. Efficient production of polymalic acid by a novel isolated Aureobasidium pullulans using metabolic intermediates and inhibitors. Appl. Biochem. Biotechnol. Doi:10.1007/s12010-018-2825-0.

    Google Scholar 

  • Zou, X., Yang, J., Tian, X., Guo, M., Li, Z., and Li, Y. 2016. Production of polymalic acid and malic acid from xylose and corncob hydrolysate by a novel Aureobasidium pullulans YJ 6–11 strain. Process Biochem. 51, 16–23.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zhiqun Liang.

Additional information

Supplemental material for this article may be found at https://doi.org/www.springerlink.com/content/120956.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zeng, W., Zhang, B., Liu, Q. et al. Analysis of the L-malate biosynthesis pathway involved in poly(β-L-malic acid) production in Aureobasidium melanogenum GXZ-6 by addition of metabolic intermediates and inhibitors. J Microbiol. 57, 281–287 (2019). https://doi.org/10.1007/s12275-019-8424-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12275-019-8424-0

Keywords

Navigation