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3 Biotech

, 9:374 | Cite as

A new strategy for the efficient production of pyocyanin, a versatile pigment, in Pseudomonas aeruginosa OG1 via toluene addition

  • Murat OzdalEmail author
Original Article

Abstract

Pseudomonas aeruginosa produce pyocyanin, which is an extracellular secondary metabolite and multifunctional pigment. In this study, the effects of several surfactants (Tween 20, Tween 80 and Triton X-100) and organic solvents (toluene and chloroform) on pyocyanin production and cell growth were investigated in submerged culture of P. aeruginosa OG1. Organic solvents were found to be more effective in the production of pyocyanin. The maximum production of pyocyanin (33 mg/L) was achieved when 0.2% toluene was added at the stationary growth phase (30 h), corresponding to significant increase of 312% compared with the control (8 mg/L). With the addition of toluene, pyocyanin production was significantly increased, but bacterial biomass reduced. Production of alkaline protease was also affected by toluene addition. It was found that the ratio of saturated/unsaturated fatty acids in the bacterial biomass significantly increased when toluene addition to the medium. This study revealed that with a novel strategy, the addition of toluene to the fermentation medium significantly increased pyocyanin production. These findings suggest that solvent-assisted fermentation strategy can be used in microbial fermentations to increase the production of biotechnological products such as industrially important pigment and enzyme. This study is a first investigation on the stimulation of pyocyanin release in the medium of P. aeruginosa cultures by the addition of toluene.

Keywords

Pseudomonas aeruginosa Pyocyanin Toluene Cell growth Fatty acids Alkaline protease 

Notes

Acknowledgements

The financial support by Department of Biology, Ataturk University, is gratefully acknowledged.

Author contributions

All the experiments were designed and executed by MO.

Compliance with ethical standards

Conflict of interest

The corresponding author states that there is no conflict of interest.

References

  1. Byreddy AR, Rao NM, Barrow CJ, Puri M (2017) Tween 80 influences the production of intracellular lipase by Schizochytrium S31 in a stirred tank reactor. Process Biochem 53:30–35.  https://doi.org/10.1016/j.procbio.2016.11.026 CrossRefGoogle Scholar
  2. Cheluvappa R (2014) Standardized chemical synthesis of Pseudomonas aeruginosa pyocyanin. MethodsX. 1:67–73.  https://doi.org/10.1016/j.mex.2014.07.001 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Essar DW, Eberly LEE, Hadero A, Crawford IP (1990) Identification and characterization of genes for a second anthranilate synthase in Pseudomonas aeruginosa: interchangeability of the two anthranilate synthases and evolutionary implications. J Bacteriol 172:884–900.  https://doi.org/10.1128/jb.172.2.884-900.1990 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Frank LH, DeMoss RD (1959) On the biosynthesis of pyocyanine. J Bacteriol 77:776–782PubMedPubMedCentralGoogle Scholar
  5. Gaur R, Khare SK (2009) Cellular response mechanisms in Pseudomonas aeruginosa PseA during growth in organic solvents. Lett Appl Microbiol 49:372–377.  https://doi.org/10.1111/j.1472-765X.2009.02671.x CrossRefPubMedGoogle Scholar
  6. Gupta A, Khare SK (2006) A protease stable in organic solvents from solvent tolerant strain of Pseudomonas aeruginosa. Bioresour Technol 97:1788–1793.  https://doi.org/10.1016/j.biortech.2005.09.006 CrossRefPubMedGoogle Scholar
  7. Hall S, McDermott C, Anoopkumar-Dukie S, McFarland AJ, Forbes A, Perkins AV, Davey AK, Chess-Williams R, Kiefel MJ, Arora D, Grant GD (2016) Cellular effects of pyocyanin, a secreted virulence factor of Pseudomonas aeruginosa. Toxins 8:236.  https://doi.org/10.3390/toxins8080236 CrossRefPubMedCentralGoogle Scholar
  8. He P, Wu S, Pan L, Sun S, Mao D, Xu C (2016) Effect of tween 80 and acetone on the secretion, structure and antioxidant activities of exopolysaccharides from Lentinus tigrinus. Food Technol Biotechnol 54:290–295.  https://doi.org/10.17113/ftb.54.03.16.4211 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Jayaseelan S, Ramaswamy D, Dharmaraj S (2014) Pyocyanin: production, applications, challenges and new insights. World J Microbiol Biotechnol 30:1159–1168.  https://doi.org/10.1007/s11274-013-1552-5 CrossRefPubMedGoogle Scholar
  10. Jeong JC, Lee IY, Kim SW, Park YH (1999) Stimulation of β-carotene synthesis by hydrogen peroxide in Blakeslea trispora. Biotechnol Lett 21:683–686.  https://doi.org/10.1023/A:1005507630470 CrossRefGoogle Scholar
  11. Kurbanoglu EB, Ozdal M, Ozdal OG, Algur OF (2015) Enhanced production of prodigiosin by Serratia marcescens MO-1 using ram horn peptone. Braz J Microbiol 46:631–637.  https://doi.org/10.1590/S1517-838246246220131143 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Laxmi M, Bhat SG (2016) Characterization of pyocyanin with radical scavenging and antibiofilm properties isolated from Pseudomonas aeruginosa strain BTRY1. 3 Biotech 6:27.  https://doi.org/10.1007/s13205-015-0350-1 CrossRefPubMedPubMedCentralGoogle Scholar
  13. Lei XY, Zhang MY, Ma YJ, Wang JW (2017) Transcriptomic responses involved in enhanced production of hypocrellin A by addition of Triton X-100 in submerged cultures of Shiraia bambusicola. J Ind Microbiol Biotechnol 44:1415–1429.  https://doi.org/10.1007/s10295-017-1965-5 CrossRefPubMedGoogle Scholar
  14. Li L, Komatsu T, Inoue A, Horikoshi K (1995) A toluene-tolerant mutant of Pseudomonas aeruginosa lacking the outer membrane protein F. Biosci Biotechnol Biochem 59:2358–2359.  https://doi.org/10.1271/bbb.59.2358 CrossRefPubMedGoogle Scholar
  15. Lim JM, Yun JW (2006) Enhanced production of exopolysaccharides by supplementation of toluene in submerged culture of an edible mushroom Collybia maculata TG-1. Process Biochem 41:1620–1626.  https://doi.org/10.1016/j.procbio.2006.03.011 CrossRefGoogle Scholar
  16. Liyama K, Takahashi E, Lee JM, Mon H, Morishita M, Kusakabe T, Yasunaga-Aoki C (2017) Alkaline protease contributes to pyocyanin production in Pseudomonas aeruginosa. FEMS Microbiol Lett 364:fnx051.  https://doi.org/10.1093/femsle/fnx051 CrossRefGoogle Scholar
  17. Lundgren BR, Thornton W, Dornan MH, Villegas-Peñaranda LR, Boddy CN, Nomura CT (2013) The gene PA2449 is essential for glycine metabolism and pyocyanin biosynthesis in Pseudomonas aeruginosa PAO1. J Bacteriol 195:2087–2100.  https://doi.org/10.1128/JB.02205-12 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Orlandi VT, Bolognese F, Chiodaroli L, Tolker-Nielsen T, Barbieri P (2015) Pigments influence the tolerance of Pseudomonas aeruginosa PAO1 to photodynamically induced oxidative stress. Microbiol 161:2298–2309.  https://doi.org/10.1099/mic.0.000193 CrossRefGoogle Scholar
  19. Özcan D, Kahraman H (2015) Pyocyanin production in the presence of calcium ion in Pseudomonas aeruginosa and recombinant bacteria. Turkish J Sci Technol 10:13–19Google Scholar
  20. Ozdal M, Ozdal OG, Algur OF (2016) Isolation and characterization of α-endosulfan degrading bacteria from the microflora of cockroaches. Pol J Microbiol 65:63–68.  https://doi.org/10.5604/17331331.1197325 CrossRefPubMedGoogle Scholar
  21. Ozdal M, Gurkok S, Ozdal OG (2017a) Optimization of rhamnolipid production by Pseudomonas aeruginosa OG1 using waste frying oil and chicken feather peptone. 3 Biotech 7:117.  https://doi.org/10.1007/s13205-017-0774-x CrossRefPubMedPubMedCentralGoogle Scholar
  22. Ozdal M, Ozdal OG, Gurkok S (2017b) Statistical optimization of beta-carotene production by Arthrobacter agilis A17 using response surface methodology and Box-Behnken design. AIP Conf Proc 1833:020101.  https://doi.org/10.1063/1.4981750 CrossRefGoogle Scholar
  23. Pacífico C, Fernandes P, de Carvalho CCCR (2018) Mycobacterial response to organic solvents and possible implications on cross-resistance with antimicrobial agents. Front Microbiol 9:961.  https://doi.org/10.3389/fmicb.2018.00961 CrossRefPubMedPubMedCentralGoogle Scholar
  24. Patil S, Nikam M, Patil H, Anokhina T, Kochetkov V, Chaudhari A (2017) Bioactive pigment production by Pseudomonas spp. MCC 3145: statistical media optimization, biochemical characterization, fungicidal and DNA intercalation-based cytostatic activity. Process Biochem 58:298–305.  https://doi.org/10.1016/j.procbio.2017.05.003 CrossRefGoogle Scholar
  25. Pierson LS, Pierson EA (2010) Metabolism and function of phenazines in bacteria: impacts on the behavior of bacteria in the environment and biotechnological processes. Appl Microbiol Biotechnol 86:1659–1670.  https://doi.org/10.1007/s00253-010-2509-3 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Rakicka M, Rywińska A, Cybulski K, Rymowicz W (2016) Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants. Braz J Microbiol 47:417–423.  https://doi.org/10.1016/j.bjm.2016.01.011 CrossRefPubMedPubMedCentralGoogle Scholar
  27. Santos IC, Chaumette A, Smuts J, Hildenbrand ZL, Schug KA (2019) Analysis of bacteria stress responses to contaminants derived from shale energy extraction. Environ Sci Process Impacts 21:269–278.  https://doi.org/10.1039/C8EM00338F CrossRefPubMedGoogle Scholar
  28. Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. MIDI Technical Note 101. MIDI Inc., NewarkGoogle Scholar
  29. Sheng L, Tang G, Su P, Zhang J, Xiao Q, Tong Q, Ma M (2016) Understanding the influence of Tween 80 on pullulan fermentation by Aureobasidium pullulans CGMCC1234. Carbohydr Polym 136:1332–1337.  https://doi.org/10.1016/j.carbpol.2015.10.058 CrossRefPubMedGoogle Scholar
  30. Sumarsih S, Nimatuzahroh F, Puspitasari M, Rusdiana M (2017) Effect of aliphatic and aromatic hydrocarbons on the oxygenase production from hydrocarbonoclastic bacteria. J Chem Technol Biotechnol 52:1062–1069Google Scholar
  31. Svenningsen NB, Pérez-Pantoja D, Nikel PI, Nicolaisen MH, de Lorenzo V, Nybroe O (2015) Pseudomonas putida mt-2 tolerates reactive oxygen species generated during matric stress by inducing a major oxidative defense response. BMC Microbiol 15:202.  https://doi.org/10.1186/s12866-015-0542-1 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Thumar JT, Singh SP (2009) Organic solvent tolerance of an alkaline protease from salt-tolerant alkaliphilic Streptomyces clavuligerus strain Mit-1. J Ind Microbiol Biotechnol 36:211–218.  https://doi.org/10.1007/s10295-008-0487-6 CrossRefPubMedGoogle Scholar
  33. Torres S, Pandey A, Castro GR (2011) Organic solvent adaptation of Gram positive bacteria: applications and biotechnological potentials. Biotechnol Adv 29:442–452.  https://doi.org/10.1016/j.biotechadv.2011.04.002 CrossRefPubMedGoogle Scholar
  34. Vinckx T, Wei Q, Matthijs S, Cornelis P (2010) The Pseudomonas aeruginosa oxidative stress regulator OxyR influences production of pyocyanin and rhamnolipids: protective role of pyocyanin. Microbiol 156:678–686.  https://doi.org/10.1099/mic.0.031971-0 CrossRefGoogle Scholar
  35. Wang Y, Zhang B, Lu L, Huang Y, Xu G (2013) Enhanced production of pigments by addition of surfactants in submerged fermentation of Monascus purpureus H1102. J Sci Food Agric 93:3339–3344.  https://doi.org/10.1002/jsfa.6182 CrossRefPubMedGoogle Scholar
  36. Wu CH, Yet-Pole I, Chiu YH, Lin CW (2014) Enhancement of power generation by toluene biodegradation in a microbial fuel cell in the presence of pyocyanin. J Taiwan Inst Chem Engrs 45:2319–2324.  https://doi.org/10.1016/j.jtice.2014.05.019 CrossRefGoogle Scholar
  37. Xu J, Du W, Zhao X, Liu D (2016) Renewable microbial lipid production from Oleaginous Yeast: some surfactants greatly improved lipid production of Rhodosporidium toruloides. World J Microbiol Biotechnol 32:107.  https://doi.org/10.1007/s11274-016-2076-6 CrossRefPubMedGoogle Scholar
  38. Zhang BB, Cheung PC (2011) A mechanistic study of the enhancing effect of Tween 80 on the mycelial growth and exopolysaccharide production by Pleurotus tuber-regium. Bioresour Technol 102:8323–8326.  https://doi.org/10.1016/j.biortech.2011.06.021 CrossRefPubMedGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Department of Biology, Science FacultyAtaturk UniversityErzurumTurkey

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