3 Biotech

, 9:5 | Cite as

Characterisation of Streptomyces violascens OC125-8 lipase for oily wastewater treatment

  • Rukiye BoranEmail author
  • Aysel Ugur
  • Nurdan Sarac
  • Ozgur Ceylan
Original Article


In this study, the lipase-producing bacterium Streptomyces violascens (GenBank number MF621564) was identified, and the extracellular S. violascens OC125-8 lipase produced by this strain was characterised for use in wastewater treatment. The lipase was partially purified by ammonium sulphate precipitation at a final yield of 3.28-fold purification and a recovery of 56%. The S. violascens OC125-8 lipase exhibited optimum catalytic activity at 40 °C and pH 8.0; it was stable at 30–40 °C with more than 86% residual activity after 1 h; it was also stable over a relatively broad pH range of pH 7.0–11.0, retaining 83.3–100% activity. Vmax and Km values were calculated as 0.61 µmol/min/mg and 0.259 mM, respectively. Enzyme activity significantly increased in the presence of Fe2+ ion but was inhibited by Ca2+, Mn2+, Cu2+ and Mg2+. The addition of a serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), strongly inhibited enzyme activity while ethylenediaminetetraacetic acid (EDTA), a metal chelating agent, had no inhibitory effect. The enzyme was fairly stable in the presence of surfactants as well as sodium perborate. Examination of commercial detergent tolerance revealed that the lipase was strongly stable in Tursil (88%), Pril (97%) and Fairy (98.5%), while the lipase was activated in Omo (113.4%) and Ariel (128.3%). Moreover, the lipase showed highest activity towards olive oil (100%), sunflower oil (90%) and burned sunflower oil (55%), while corn oil (44%) and burned olive oil (15%) were less hydrolysed by the enzyme. These properties demonstrate that S. violascens OC125-8 lipase is an ideal choice for oily wastewater management.


Streptomyces violascens Lipase Characterisation Wastewater Oil degradation 



We would like to express our gratitude to Dr. Şimşek for his kind help during the identification of bacterial strain.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


  1. Abd El-Gawad HS (2014) Oil and grease removal from industrial wastewater using new utility approach. Adv Environ Chem 2014:1–6. CrossRefGoogle Scholar
  2. Ascherl F (2014) Borates in detergency. Accessed 13 Oct 2017
  3. Ayaz B, Ugur A, Boran R (2015) Purification and characterization of organic solvent-tolerant lipase from Streptomyces sp. OC119-7 for biodiesel production. Biocatal Agric Biotechnol 4(1):103–108. CrossRefGoogle Scholar
  4. Ben Elhoul MB, Jaouadi NZ, Rekika H, Bejar W, Touioui SB, Hmidi M, Badis A, Bejar S, Jaouadi B (2015) A novel detergent-stable solvent-tolerant serine thiol alkaline protease from Streptomyces koyangensis TN650. Int J Biol Macromol 79:871–882. CrossRefPubMedGoogle Scholar
  5. Bentley SD, Chater KF, Cerdeño-Tárraga AM, Challis GL, Thomson NR, James KD, Harris DE, Quail MA, Kieser H, Harper D, Bateman A, Brown S, Chandra G, Chen CW, Collins M, Cronin A, Fraser A, Goble A, Hidalgo J, Hornsby T, Howarth S, Huang CH, Kieser T, Larke L, Murphy L, Oliver K, O’Neil S, Rabbinowitsch E, Rajandream MA, Rutherford K, Rutter S, Seeger K, Saunders D, Sharp S, Squares R, Squares S, Taylor K, Warren T, Wietzorrek A, Woodward J, Barrell BG, Parkhill J, Hopwood DA (2002) Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417:141–147. CrossRefGoogle Scholar
  6. Bhumibhamon O, Koprasertsak A, Funthong S (2002) Biotreatment of high fat and oil wastewater by lipase producing microorganisms. Nat Sci 36:261–267Google Scholar
  7. Bielen A, C´etkovic´ H, Long PF, Schwab H, Abramic´ M, Vujaklija D (2009) The SGNH-hydrolase of Streptomyces coelicolor has (aryl)esterase and a true lipase activity. Biochimie 91:390–400. CrossRefPubMedGoogle Scholar
  8. Boran R, Ugur A (2010) Partial purification and characterization of the organic-solvent-tolerant lipase produced by Pseudomonas fluorescens RB02-3 isolated from milk. Prep Biochem Biotechnol 40(4):229–241. CrossRefPubMedGoogle Scholar
  9. Bradford MM (1976) A rapid and sensitive for the quantitation of microgram quantititesof protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  10. Burgess JE, Pletschke BI (2008) Hydrolytic enzymes in sewage sludge treatment: a mini-review. Water SA 34:343–350Google Scholar
  11. Clark LC, Seipke RF, Prieto P, Willemse J, van Wezel GP, Hutchings MI, Hoskisson PA (2013) Mammalian cell entry genes in Streptomyces may provide clues to the evolution of bacterial virulence. Sci Rep 3 (1109):1–8. CrossRefGoogle Scholar
  12. Colla LM, Rizzardi J, Pinto MH, Reinehr CO, Bertolin TE, Costa JAV (2010) Simultaneous production of lipases and bio-surfactants by submerged and solid-state bioprocesses. Bioresour Technol 101:8308–8314. CrossRefPubMedGoogle Scholar
  13. Dandavate V, Jinjala J, Keharia H, Madamwar D (2009) Production, partial purification and characterization of organic solvent tolerant lipase from Burkholderia multivorans V2 andits application for ester synthesis. Bioresour Technol 100(13):3374–3381. CrossRefPubMedGoogle Scholar
  14. Habbeche A, Saoudi B, Jaouadi B, Haberra S, Kerouaz B, Boudelaa M, Badis A, Ladjama A (2014) Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost. J Biosci Bioeng 117:413–421. CrossRefPubMedGoogle Scholar
  15. Jamie A, Alshami AS, Maliabari ZO, Ateih MA, Al Hamouza OCS (2016) Immobilization and enhanced catalytic activity of lipase on modified MWCNT for oily wastewater treatment. Environ Prog Sustain Energy 35:1441–1449. CrossRefGoogle Scholar
  16. Karhu M (2015) Treatment and characterisation of oily wastewaters, PhD, Faculty of Technology, University of OuluGoogle Scholar
  17. Leal MCMR, Freire DMG, Cammarota MC, Sant’Anna GL Jr (2006) Effect of enzymatic hydrolysis on anaerobic treatment of dairy wastewater. Process Biochem 41(5):1173–1178. CrossRefGoogle Scholar
  18. Lesuisse E, Schanck K, Colson C (1993) Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme. Eur J Biochem 216:155–160CrossRefGoogle Scholar
  19. Mander P, Cho SS, Simkhada JR, Choi YH, Park DJ, Ha JW, Yoo JC (2012) An organic solvent-tolerant alkaline lipase from Streptomyces sp. CS268 and its application in biodiesel production. Biotechnol Bioprocess Eng 17(1):67–75. CrossRefGoogle Scholar
  20. Mander P, Yoo H-Y, Kim SW, Choi YH, Cho SS, Yoo JC (2014) Transesterification of waste cooking oil by an organic solvent-tolerant alkaline lipase from Streptomyces sp. CS273. Appl Biochem Biotechnol 172(3):1377–1389. CrossRefPubMedGoogle Scholar
  21. Meng Y, Luan F, Yuan H, Chen X, Li X (2017) Enhancing anaerobic digestion performance of crude lipid in food waste by enzymatic pretreatment. Bioresour Technol 224:48–55. CrossRefPubMedGoogle Scholar
  22. Mugdha A, Usha M (2012) Enzymatic treatment of wastewater containing dyestuffs using different delivery systems. Sci Revs Chem Commun 2(1):31–40Google Scholar
  23. Nelson C, Cox M (2004) Principles of biochemistry, 4th edn. W. H. Freeman, New YorkGoogle Scholar
  24. Nzila A, Thukair A, Sankara S, AbdurRazzak S (2017) Characterization of aerobic oil and grease-degrading bacteria in wastewater. Environ Technol 38(6):661–670. CrossRefPubMedGoogle Scholar
  25. Ogino H, Nakagawa S, Shinya K, Muto T, Fujimura N, Yasuda M, Ishikawa H (2000) Purification and characterization of organic solvent-stable lipase from organic solvent-tolerant Pseudomonas aeruginosa LST-03. J Biosci Bioeng 89:451–457. CrossRefPubMedGoogle Scholar
  26. Rajanikanth A, Damodharam T (2017) Partial purification and characterization of detergent compatible lipase from marine Streptomyces fungicidicus RPBS-A4 for application in oil based stain removal. IJCAR 6(6):4345–4351. CrossRefGoogle Scholar
  27. Rigo E, Rigoni RE, Lodea P, Oliveira DD, Freire DMG, Luccio MD (2008) Application of different lipases as pretreatment in anaerobic treatment of wastewater. J Environ Eng Ecol Sci 25(9):1243–1248. CrossRefGoogle Scholar
  28. Rosa DR, Duarte ICS, Katia Saavedra N, Varesche MB, Zaiat M, Cammarota MC, Freire DMG (2009) Performance and molecular evaluation of an anaerobic system with suspended biomass for treating wastewater with high fat content after enzymatic hydrolysis. Bioresour Technol 100:6170–6176. CrossRefPubMedGoogle Scholar
  29. Roskill (2002) The economics of Boron, 10th edn. Roskill Information Services, LondonGoogle Scholar
  30. Sarac N, Ugur A, Sen B (2017) A green alternative for biodiesel production: transesterification with Streptomyces sp. AU-1 lipase. Rom Biotechnol Lett 22(6):13060–13067Google Scholar
  31. Sen B, Sarac N, Ugur A (2016) Partial purification, characterization and biodiesel application of Streptomyces lienomycini lipase. Rom Biotechnol Lett 22(6):12103–12109Google Scholar
  32. Tang L, Xia Y, Wu X, Chen X, Zhang X, Li H (2017) Screening and characterization of a novel thermostable lipase with detergent-additive potential from the metagenomic library of a mangrove soil. Gene 625:64–71. CrossRefPubMedGoogle Scholar
  33. Ugur A, Boran R (2014) Production and characterization of a cold-active and n-hexane activated lipase from a newly isolated Serratia grimesii RB06-22. Biocatal Biotransform 32(4):222–230. CrossRefGoogle Scholar
  34. Ugur A, Sarac N, Boran R, Ayaz B, Ceylan O, OkmenG (2014) New lipase for biodiesel production: Partial purification and characterization of LipSB 25-4. ISRN Biochem. (ID 289749)CrossRefGoogle Scholar
  35. Ungcharoenwiwat P, Kittikun AH (2015) Purification and characterization of lipase from Burkholderia sp. EQ3 isolated from wastewater from a canned fish factory and its application for the synthesis of wax esters. J Mol Catal B Enzym 115:96–104. CrossRefGoogle Scholar
  36. Valladão ABG, Freire DMG, Cammarota MC (2007) Enzymatic pre-hydrolysis—applied to the anaerobic treatment of effluents from poultry slaughterhouses. Int Biodeterior Biodegrad 60:219–225. CrossRefGoogle Scholar
  37. Van Oort MG, Deveer AMTJ, Dijkman R, Tjeenk ML, Verheij HM, De Haas GH, Wenzig E, Goetz F (1989) Purification and substrate specificity of Staphylococcus hyicus lipase. Biochemistry 28:9278–9285. CrossRefPubMedGoogle Scholar
  38. Yuan D, Lan D, Xin R, Yang B, Wang Y (2016) Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007. Biotechnol Appl Biochem 63:41–50. CrossRefPubMedGoogle Scholar
  39. Zhang Y, Meng K, Wang Y, Luo H, Wu PP, Fan N, Li Y, Yao J B (2008) A novel proteolysis-resistant lipase from keratinolytic Streptomyces fradiae var. k11. Enzyme Microb Technol 42:346–352. CrossRefGoogle Scholar
  40. Zhang Y, Cui B, Wang S, Chu Z, Fan X, Hua Y, Lan Y (2012) Relation between enzyme activity of sediments and lake eutrophication in grass-type lakes in North China. Clean-Soil Air Water 40(10):1145–1153. CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Medical Laboratory Program, Department of Medical Services and Techniques, Vocational School of Health ServiceAksaray UniversityAksarayTurkey
  2. 2.Section of Medical Microbiology, Department of Basic Sciences, Faculty of DentistryGazi UniversityAnkaraTurkey
  3. 3.Department of Biology, Faculty of ScienceMuğla Sıtkı Koçman UniversityMuglaTurkey
  4. 4.Food Quality Control and Analysis Program, Ula Ali Koçman Vocational SchoolMuğla Sıtkı Koçman UniversityMuglaTurkey

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