Advertisement

3 Biotech

, 8:370 | Cite as

Fungal biotransformation of synthetic levodopa to stable dopamine in l-ascorbate-mediated aerobic-thermophilic biochemical process

  • Sikander Ali
Original Article

Abstract

In the present study, fungal biotransformation of synthetic levodopa to stable dopamine in an l-ascorbate-mediated thermophilic-aerobic biochemical reaction was investigated. A mutant strain of Aspergillus oryzae EMS-6 was used for the preparation of mycelial biomass. The mutant was previously developed through EMS-induced mutagenesis and repressed against l-cysteine HCl. Growth parameters such as rate of cultivation (48 h), initial pH (6) and incubation temperature (30 °C) supported 18.84 g/l biomass with 23 g/l glucose consumption. Thermophilic behaviour of culture was observed at 25–40 °C. Kinetic variables notably µ = 0.385 /h and Qs, exhibited consistent growth pattern. Biochemical reactions were performed aerobically using mycelial biomass as the source of enzyme ‘tyrosinase’ in a digital hotplate equipped with magnetic stirrers. The reaction conditions included 5 mg/ml biomass and 2.5 mg/ml levodopa as basal substrate in a thermophilic reaction of 25 min duration acidified with l-ascorbic acid. TLC and HPLC analysis of reaction mixture confirmed the presence of levodopa and dopamine using a CN-9dth (R) column. Activation enthalpy and entropy of dopa decarboxylase (DDC) and its thermal inactivation showed an improved biotransformation of levodopa to dopamine at the optimal temperature (30 °C) as compared to other temperatures being employed. Overall, 3.68 mg/ml dopamine (4.55 mg/ml proteins) synthesis from 2.38 mg/ml levodopa was accomplished. The enhancement in metabolic activity of the mutant strain is ~ 2.75-fold improved when compared to the unoptimized reaction conditions, which is highly significant (HS) indicating an eco-commercially viable (LSD ~ 0.412) bioprocess.

Keywords

Aspergillus oryzae Levodopa Dopamine Dopa decarboxylase Biotransformation l-ascorbate 

Notes

Compliance with ethical standards

Conflict of interest

The author declares that there is no conflict of interest.

References

  1. Aiba S, Humphrey AE, Millis NF (1973) Biochemical engineering, 2nd edn. Academic Press, New York, pp 92–127.  https://doi.org/10.1002/jobm Google Scholar
  2. Ali S, Haq I, Qadeer MA, Rajoka MI (2005) Double mutant of Aspergillus oryzae for improved production of 3,4-dihydroxy phenyl-l-alanine from l-tyrosine. Biotechnol Appl Biochem 42:143–149.  https://doi.org/10.1042/BA20040180 CrossRefGoogle Scholar
  3. Arnow LE (1973) Colorimetric determination of the components of l-3,4-dihydroxy phenylalanine in tyrosine mixtures. J Biochem 88:531Google Scholar
  4. Bertoldi M, Voltattorni CB (2000) Reaction of dopa decarboxylase with l-aromatic amino acids under aerobic and anaerobic condition. Biochem J 352:533–538CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid method for the quantification of program quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Charteris A, John RA (1975) An investigation of the assay of dopamine using trinitrobenzenesulphonic acid. Anal Biochem 66:365–371CrossRefGoogle Scholar
  7. Christensen G (1972) Effects of metal cations and other chemicals upon the in vitro activity of two enzymesin the blood plasma of the white sucker Catostomus commersoni. Chem Biol Interact 4:351–361CrossRefGoogle Scholar
  8. Converti A, Dominguez JM (2001) Influence of temperature and pH on xylitol production from xylose by Debarryomyces hansenii. Biotechnol Bioeng 75:39–45CrossRefGoogle Scholar
  9. Felger JC, Miller AH (2012) Cytokine effects on the basal ganglia and dopamine function: the subcortical source of inflammatory malaise. Front Neuroendocrinol 33:315–327.  https://doi.org/10.1016/j.yfrne.2012.09.003 CrossRefGoogle Scholar
  10. Felger JC, Hernandez CR, Miller AH (2015) Levodopa reverses cytokine-induced reductions in striatal dopamine release. Int J Neuropsychopharmacol 18:84–96.  https://doi.org/10.1093/ijnp/pyu084 CrossRefGoogle Scholar
  11. Haneda Y, Tsuji FI, Sugiyama N (1969) Luminescent systems in apogonid fishes from the Philippines. Science 165(3889):188–190CrossRefGoogle Scholar
  12. Haneda K, Watanabe S, Takeda P (1973) l-DOPA by microorganisms. J Ferment Technol 51:398–406Google Scholar
  13. Haq I, Ali S, Qadeer MA (2002) Biosynthesis of l-Dopa by Aspergillus oryzae. Bioresour Technol 85:25–29.  https://doi.org/10.1016/S0960-8524(02)00060-3 CrossRefGoogle Scholar
  14. Hodgetts RB, O’Keefe SL (2006) Dopa decarboxylase: a model gene-enzyme system for studying development, behaviour and systematics. Annu Rev Entomol 51:259–284.  https://doi.org/10.1146/annurev.ento.51.110104.151143 CrossRefGoogle Scholar
  15. Hyland K, Surtees RAH, Rodeck C, Clayton PT (1992) Aromatic l-amino acid decarboxylase deficiency: clinical features, diagnosis and treatmentof a new born error of neurotransmitter amine synthesis. Neurol 42:1980–1988CrossRefGoogle Scholar
  16. Kaljunen H, Gasparetti C, Kruus K, Rouvinen J, Hakulinen N (2011) Crystallization and preliminary X-ray analysis of Aspergillus oryzae catechol oxidase. Acta Crystallogr Sect F Struct Biol Cryst Commun 67:672–674.  https://doi.org/10.1107/S1744309107060721 CrossRefGoogle Scholar
  17. Kanteev M, Goldfeder M, Fishman A (2015) Structure-function correlations in tyrosinases. Protein Sci 24:1360–1369.  https://doi.org/10.1002/pro.2734 CrossRefGoogle Scholar
  18. Li J, Christensen BM (1993) Identification of products and intermediates during l-dopa oxidation to dopachrome using pressure liquid chromatography. J LIq Chromatogr 16:1117–1133CrossRefGoogle Scholar
  19. Minelli A, Charteris AT, Voltattorni CB, John RA (1979) Reactions of Dopa (3,4-dihydroxyphenylalanine) decarboxylase with Dopa. Biochem J 183:2843–2851CrossRefGoogle Scholar
  20. Nakamura M, Nakajima T, Ohba Y, Yamauchi S, Lee BR, Ichishima E (2000) Identification of copper ligands in Aspergillus oryzae tyrosinase by site-directed mutagenesis. Biochem J 350:537–545CrossRefGoogle Scholar
  21. Piccini P, Brooks DJ, Bjorklund A, Gunn RN, Grasby PM, Rimoldi O, Brundin P, Hagell P, Rehnrona S, Widner H, Lindwall O (1999) Dopamine release from nigral transplants visualized in vivo in a Parkinson’s patient. Nat Neurosci 2:1137–1140CrossRefGoogle Scholar
  22. Pirt SJ (1975) Principles of cell cultivation. Blackwells Scientific Corporation, LondonGoogle Scholar
  23. Pontecarvo J, Roper JA, Hemmous LM, Buftan W (1969) Basic techniques of UV-irradiation and chemical mutation. Adv Genet 5:141–183Google Scholar
  24. Raju BG, Rao GH, Ayyanna C (1993) Bioconversion of tyrosine to melanin using mycelia of Aspergillus spp. CBS Publishers, VisakhapatnamGoogle Scholar
  25. Salinas AG, Davis MI, Lovinger DM, Mateo Y (2016) Dopamine dynamics and cocaine sensitivity differ between striosome and matrix compartments of the striatum. Neuropharmacol 108:275–283.  https://doi.org/10.1016/j.neuropharm.2016.03.049 CrossRefGoogle Scholar
  26. Sherald AF, Sparrow JC, Wright TRF (1973) A spectrophotometric assay for Drosophila dopa decarboxylase. Anal Biochem 56:300–305CrossRefGoogle Scholar
  27. Sirivelu MP, Mohankumar SMJ, Wagner JG, Harkema JR, Mohankumar PS (2006) Activation of the stress axis and neurochemical alterations in specific brain areas by concentrated ambient particle exposure with concomitant allergic airway disease. Environ Health Perspect 114:870–874CrossRefGoogle Scholar
  28. Snedecor GW, Cochran WG (1980) Statistical methods, 7th edn. Ames, Iowa state University Press, Iowa.Google Scholar
  29. Tang MC, Zou Y, Watanabe K, Walsh CT, Tang Y (2017) Oxidative cyclization in natural product biosynthesis. Chem Rev 117:5226–5333.  https://doi.org/10.1021/acs.chemrev.6b00478 CrossRefGoogle Scholar
  30. Vaillancourt DE, Schonfeld D, Kwak Y, Bohnen NI, Seidler R (2013) Dopamine overdose hypothesis: evidence and clinical implications. Mov Disord 28:43–57.  https://doi.org/10.1002/mds.25687 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Ind. Biotechnology (IIB)GCULahorePakistan

Personalised recommendations