Research Article
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Production and Activity Characterization of Lipase from Bacillus flexus InaCC-B486

Year 2024, , 397 - 404, 15.05.2024
https://doi.org/10.18596/jotcsa.1333916

Abstract

Lipases are widely used in a variety of industries, both to develop products and to improve process efficiency. The need for lipase increased along with the wider application of this enzyme. Therefore, studies related to the search for potential lipase-producing microbes that answer the needs of the industry are required to be carried out continuously. Enzymes produced by microbes are preferred because they can be produced quickly compared to other sources. Bacillus flexus InaCC-B486 was used to produce lipase in this study with olive oil as substrate. This research aimed to observe the production of lipase from B. flexus InaCC-B486 and characterize its activity. The result shows that the production of B. flexus InaCC-B486 lipase was optimal at day-4 which was 11.983 ± 0.101 U/mL. The activity of B. flexus InaCC-B486 lipase was optimal at an incubation time of 15 minutes (2.810 U/mL), pH of 8.0 (3.173 U/mL), and a temperature of 35 oC (3.173 U/mL). These findings can be used for further applications, both in research and industry, that use B. flexus InaCC-B486 as a resource for lipase production or any related applications.

Supporting Institution

National Research and Innovation Agency of Indonesia

References

  • 1. Coelho ALS, Orlandelli RC. Immobilized microbial lipases in the food industry: a systematic literature review. Crit Rev Food Sci Nutr. 2021 May 31;61(10):1689–703. Available from: <URL>.
  • 2. Negi S. Lipases: A Promising Tool for Food Industry. In: Parameswaran B, Varjani S, Raveendran S, editors. Green Bio-processes: Enzymes in Industrial Food Processing. Singapore: Springer Singapore; 2019. p. 181–98. Available from: <URL>.
  • 3. Bharathi D, Rajalakshmi G. Microbial lipases: An overview of screening, production and purification. Biocatal Agric Biotechnol. 2019;22:101368. Available from: <URL>.
  • 4. Ismail AR, Baek KH. Lipase immobilization with support materials, preparation techniques, and applications: Present and future aspects. Int J Biol Macromol. 2020;163:1624–39. Available from: <URL>.
  • 5. Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, et al. Applications of microbial enzymes in food industry. Vol. 56, Food Technology and Biotechnology. University of Zagreb; 2018. p. 16–30. Available from: <URL>
  • 6. Guerrand D. Lipases industrial applications: Focus on food and agroindustries. In: OCL - Oilseeds and fats, Crops and Lipids. EDP Sciences; 2017. Available from: <URL>
  • 7. Pessione E. The Less Expensive Choice: Bacterial Strategies to Achieve Successful and Sustainable Reciprocal Interactions. Vol. 11, Frontiers in Microbiology. Frontiers Media S.A.; 2021. Available from: <URL>
  • 8. Daroonpunt R, Saeng-in P, Tanasupawat S. Identification and lipolytic activity of Bacillus and Staphylococcus strains from shrimp paste (Ka-pi). J Appl Pharm Sci. 2019 Apr 1;9(4):24–9. Available from: <URL>.
  • 9. Mukesh kumar DJ, Andal Priyadharshini D, Suresh K, Saranya G, Rajendran K, Kalaichelven P. Production, purification and characterization of α-amylase and alkaline protease by Bacillus sp. HPE 10 in a concomitant production medium. Asian Journal of Plant Science and Research. 2012;2(3):376–82.
  • 10. Roy P, Chatterjee S, Saha NC, Gantait VV. Characterization of a Starch Hydrolysing Bacillus flexus U8 Isolated from Rhizospheric Soil of the Paddy Plants. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2020;90(5):1075–81. Available from: <URL>.
  • 11. Tambekar DH, Tambekar SD, Jadhav AS, Kharat PA. Alkalıphılıc Bacıllus flexus: A potentıal source of lıpase producer for ındustrıal and medıcal applıcatıon. Int J Pharm Sci Res. 2017;8(10):11. Available from: <URL>.
  • 12. Setyati WA, Martani E, Triyanto, Subagiyo, Zainuddin M. Selection, identification and optimization of the growth water probiotic consortium of mangrove ecosystems as bioremediation and biocontrol in shrimp ponds. Indonesian Fisheries Processing Journal. 2014;17(3):243–53. Available from: <URL>
  • 13. Sarastiti S, Suminto, Sarjito. Molecular identification bacteria as probiotic candidates isolated from intestinal tract of vaname shrimp (Litopenaeus vannamei) collected from Subang, West Java. Journal of Coastal and Marine Resources Management. 2020;4(1):9–15.
  • 14. Soleymani S, Alizadeh H, Mohammadian H, Rabbani E, Moazen F, Sadeghi HM, et al. Efficient Media for High Lipase Production: One Variable at a Time Approach. Vol. 9. Available from: <URL>
  • 15. Picazo-Espinosa R. Lipase activity assay: improvements to the classical p-nitrophenol palmitate spectrophotometrical method. In: 41st International Conference of the Slovak Society of Chemical Engineering (SSCHE). Tatranské Matliare; 2014. Available from: <URL>
  • 16. Zarevúcka M. Olive Oil as Inductor of Microbial Lipase. In: Boskou D, editor. Olive Oil - Constituents, Quality, Health Properties and Bioconversion. Rijeka: InTech; 2012 [cited 2023 Jul 17]. p. 457–70. Available from: <URL>
  • 17. Abbas N, Javed J, Abbas Z, Choudry S, Ali S. Lipase production from Bacillus subtilis using various agricultural waste. International Journal of Advanced engineering, Management and Science. 2017;3(5):405–9. Available from: <URL>
  • 18. Demirkan E, Aybey Çetinkaya A, Abdou M. Lipase from new isolate Bacillus cereus ATA179: Optimization of production conditions, partial purification, characterization and its potential in the detergent industry. Turkish Journal of Biology. 2021;45(3):287–300. Available from: <URL>
  • 19. Barik A, Sen SK, Rajhans G, Raut S. Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4. Basheer C, editor. Int J Anal Chem. 2022;2022:6403090. Available from: <URL>.
  • 20. Bora L, Bora M. Optimization of extracellular thermophilic highly alkaline lipase from thermophilic Bacillus sp. isolated from hotspring of Arunachal Pradesh, India. Brazilian Journal of Microbiology. 2012;43:30–42. Available from: <URL>
  • 21. Niyonzima FN, More S. Biochemical properties of the alkaline lipase of Bacillus flexus XJU-1 and its detergent compatibility. Biologia (Bratisl). 2014;69(9):1108–17. Available from: <URL>.
  • 22. Jaeger KE, Kovacic F. Determination of Lipolytic Enzyme Activities. In: Filloux A, Ramos JL, editors. Pseudomonas Methods and Protocols. New York, NY: Springer New York; 2014. p. 111–34. Available from: <URL>.
  • 23. Vardar-Yel N. Investigation of the activity of lipase variants on different 4-nitrophenyl esters by spectrophotometric assay. Caucasian Journal of Science. 2021 Dec 31;8(2):292–303. Available from: <URL>.
  • 24. Cheng M, Angkawidjaja C, Koga Y, Kanaya S. Requirement of lid2 for interfacial activation of a family I.3 lipase with unique two lid structures. FEBS Journal. 2012 Oct;279(19):3727–37. Available from: <URL>.
  • 25. Golaki BP, Aminzadeh S, Karkhane AA, Yakhchali B, Farrokh P, Khaleghinejad SH, et al. Cloning, expression, purification, and characterization of lipase 3646 from thermophilic indigenous Cohnella sp. A01. Protein Expr Purif. 2015 May 1;109:120–6. Available from: <URL>.
  • 26. Zhang W, Yang H, Liu W, Wang N, Yu X. Improved performance of magnetic cross-linked lipase aggregates by interfacial activation: A robust and magnetically recyclable biocatalyst for transesterification of jatropha oil. Molecules. 2017 Dec 1;22(12). Available from: <URL>.
  • 27. Sooch BS, Singh Kauldhar B. Influence of Multiple Bioprocess Parameters on Production of Lipase from Pseudomonas sp. BWS-5. Braz Arch Biol Technol. 2013;56(5):711–21. Available from: <URL>.
  • 28. Gusniah A, Veny H, Hamzah F. Activity and stability of immobilized lipase for utilization in transesterification of waste cooking oil. Bull Chem React Eng Catal. 2020 Apr 1;15(1):242–52. Available from: <URL>.
  • 29. Taskin M, Ucar MH, Unver Y, Kara AA, Ozdemir M, Ortucu S. Lipase production with free and immobilized cells of cold-adapted yeast Rhodotorula glutinis HL25. Biocatal Agric Biotechnol. 2016 Oct 1;8:97–103. Available from: <URL>.
  • 30. Turati DFM, Almeida AF, Terrone CC, Nascimento JMF, Terrasan CRF, Fernandez-Lorente G, et al. Thermotolerant lipase from Penicillium sp. section Gracilenta CBMAI 1583: Effect of carbon sources on enzyme production, biochemical properties of crude and purified enzyme and substrate specificity. Biocatal Agric Biotechnol. 2019;17:15–24. Available from: <URL>.
  • 31. Akhter K, Karim I, Aziz B, Bibi A, Khan J, Akhtar T. Optimization and characterization of alkaliphilic lipase from a novel Bacillus cereus NC7401 strain isolated from diesel fuel polluted soil. PLoS One. 2022 Aug 30;17(8):e0273368. Available from: <URL>.
  • 32. Emtenani S, Asoodeh A, Emtenani S. Molecular cloning of a thermo-alkaliphilic lipase from Bacillus subtilis DR8806: Expression and biochemical characterization. Process Biochem. 2013;48(11):1679–85. Available from: <URL>
  • 33. Salihu A, Alam MdZ. Solvent tolerant lipases: A review. Process Biochemistry [Internet]. 2015;50(1):86–96. Available from: <URL>.
  • 34. Hu J, Cai W, Wang C, Du X, Lin J, Cai J. Purification and characterization of alkaline lipase production by Pseudomonas aeruginosa HFE733 and application for biodegradation in food wastewater treatment. Biotechnol Biotechnol Equip. 2018 May 4;32(3):583–90. Available from: <URL>.
  • 35. Huang J, Yang Z, Zhu R, Qian X, Wang Y, Li Y, et al. Efficient heterologous expression of an alkaline lipase and its application in hydrolytic production of free astaxanthin. Biotechnol Biofuels. 2018;11(1):181. Available from: <URL>.
  • 36. Robinson PK. Enzymes: principles and biotechnological applications. Essays Biochem. 2015 Nov 15;59:1–41. Available from: <URL>.
  • 37. Sharma P, Sharma N, Pathania S, Handa S. Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry. J Genet Eng Biotechnol. 2017;15(2):369–77. Available from: <URL>.
  • 38. Bora L, Gohain D, Das R. Recent advances in production and biotechnological applications of thermostable and alkaline bacterial lipases. J Chem Technol Biotechnol. 2013 Nov 1;88(11):1959–70. Available from: <URL>.
  • 39. Kumar V, Yedavalli P, Gupta V, Rao NM. Engineering lipase A from mesophilic Bacillus subtilis for activity at low temperatures. Protein Eng Des Sel. 2014 Mar;27(3):73–82. Available from: <URL>.
  • 40. Fojan P, Jonson PH, Petersen MTN, Petersen SB. What distinguishes an esterase from a lipase: A novel structural approach. Biochimie. 2000;82:1033–41. Available from: <URL>.
Year 2024, , 397 - 404, 15.05.2024
https://doi.org/10.18596/jotcsa.1333916

Abstract

References

  • 1. Coelho ALS, Orlandelli RC. Immobilized microbial lipases in the food industry: a systematic literature review. Crit Rev Food Sci Nutr. 2021 May 31;61(10):1689–703. Available from: <URL>.
  • 2. Negi S. Lipases: A Promising Tool for Food Industry. In: Parameswaran B, Varjani S, Raveendran S, editors. Green Bio-processes: Enzymes in Industrial Food Processing. Singapore: Springer Singapore; 2019. p. 181–98. Available from: <URL>.
  • 3. Bharathi D, Rajalakshmi G. Microbial lipases: An overview of screening, production and purification. Biocatal Agric Biotechnol. 2019;22:101368. Available from: <URL>.
  • 4. Ismail AR, Baek KH. Lipase immobilization with support materials, preparation techniques, and applications: Present and future aspects. Int J Biol Macromol. 2020;163:1624–39. Available from: <URL>.
  • 5. Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, et al. Applications of microbial enzymes in food industry. Vol. 56, Food Technology and Biotechnology. University of Zagreb; 2018. p. 16–30. Available from: <URL>
  • 6. Guerrand D. Lipases industrial applications: Focus on food and agroindustries. In: OCL - Oilseeds and fats, Crops and Lipids. EDP Sciences; 2017. Available from: <URL>
  • 7. Pessione E. The Less Expensive Choice: Bacterial Strategies to Achieve Successful and Sustainable Reciprocal Interactions. Vol. 11, Frontiers in Microbiology. Frontiers Media S.A.; 2021. Available from: <URL>
  • 8. Daroonpunt R, Saeng-in P, Tanasupawat S. Identification and lipolytic activity of Bacillus and Staphylococcus strains from shrimp paste (Ka-pi). J Appl Pharm Sci. 2019 Apr 1;9(4):24–9. Available from: <URL>.
  • 9. Mukesh kumar DJ, Andal Priyadharshini D, Suresh K, Saranya G, Rajendran K, Kalaichelven P. Production, purification and characterization of α-amylase and alkaline protease by Bacillus sp. HPE 10 in a concomitant production medium. Asian Journal of Plant Science and Research. 2012;2(3):376–82.
  • 10. Roy P, Chatterjee S, Saha NC, Gantait VV. Characterization of a Starch Hydrolysing Bacillus flexus U8 Isolated from Rhizospheric Soil of the Paddy Plants. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2020;90(5):1075–81. Available from: <URL>.
  • 11. Tambekar DH, Tambekar SD, Jadhav AS, Kharat PA. Alkalıphılıc Bacıllus flexus: A potentıal source of lıpase producer for ındustrıal and medıcal applıcatıon. Int J Pharm Sci Res. 2017;8(10):11. Available from: <URL>.
  • 12. Setyati WA, Martani E, Triyanto, Subagiyo, Zainuddin M. Selection, identification and optimization of the growth water probiotic consortium of mangrove ecosystems as bioremediation and biocontrol in shrimp ponds. Indonesian Fisheries Processing Journal. 2014;17(3):243–53. Available from: <URL>
  • 13. Sarastiti S, Suminto, Sarjito. Molecular identification bacteria as probiotic candidates isolated from intestinal tract of vaname shrimp (Litopenaeus vannamei) collected from Subang, West Java. Journal of Coastal and Marine Resources Management. 2020;4(1):9–15.
  • 14. Soleymani S, Alizadeh H, Mohammadian H, Rabbani E, Moazen F, Sadeghi HM, et al. Efficient Media for High Lipase Production: One Variable at a Time Approach. Vol. 9. Available from: <URL>
  • 15. Picazo-Espinosa R. Lipase activity assay: improvements to the classical p-nitrophenol palmitate spectrophotometrical method. In: 41st International Conference of the Slovak Society of Chemical Engineering (SSCHE). Tatranské Matliare; 2014. Available from: <URL>
  • 16. Zarevúcka M. Olive Oil as Inductor of Microbial Lipase. In: Boskou D, editor. Olive Oil - Constituents, Quality, Health Properties and Bioconversion. Rijeka: InTech; 2012 [cited 2023 Jul 17]. p. 457–70. Available from: <URL>
  • 17. Abbas N, Javed J, Abbas Z, Choudry S, Ali S. Lipase production from Bacillus subtilis using various agricultural waste. International Journal of Advanced engineering, Management and Science. 2017;3(5):405–9. Available from: <URL>
  • 18. Demirkan E, Aybey Çetinkaya A, Abdou M. Lipase from new isolate Bacillus cereus ATA179: Optimization of production conditions, partial purification, characterization and its potential in the detergent industry. Turkish Journal of Biology. 2021;45(3):287–300. Available from: <URL>
  • 19. Barik A, Sen SK, Rajhans G, Raut S. Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4. Basheer C, editor. Int J Anal Chem. 2022;2022:6403090. Available from: <URL>.
  • 20. Bora L, Bora M. Optimization of extracellular thermophilic highly alkaline lipase from thermophilic Bacillus sp. isolated from hotspring of Arunachal Pradesh, India. Brazilian Journal of Microbiology. 2012;43:30–42. Available from: <URL>
  • 21. Niyonzima FN, More S. Biochemical properties of the alkaline lipase of Bacillus flexus XJU-1 and its detergent compatibility. Biologia (Bratisl). 2014;69(9):1108–17. Available from: <URL>.
  • 22. Jaeger KE, Kovacic F. Determination of Lipolytic Enzyme Activities. In: Filloux A, Ramos JL, editors. Pseudomonas Methods and Protocols. New York, NY: Springer New York; 2014. p. 111–34. Available from: <URL>.
  • 23. Vardar-Yel N. Investigation of the activity of lipase variants on different 4-nitrophenyl esters by spectrophotometric assay. Caucasian Journal of Science. 2021 Dec 31;8(2):292–303. Available from: <URL>.
  • 24. Cheng M, Angkawidjaja C, Koga Y, Kanaya S. Requirement of lid2 for interfacial activation of a family I.3 lipase with unique two lid structures. FEBS Journal. 2012 Oct;279(19):3727–37. Available from: <URL>.
  • 25. Golaki BP, Aminzadeh S, Karkhane AA, Yakhchali B, Farrokh P, Khaleghinejad SH, et al. Cloning, expression, purification, and characterization of lipase 3646 from thermophilic indigenous Cohnella sp. A01. Protein Expr Purif. 2015 May 1;109:120–6. Available from: <URL>.
  • 26. Zhang W, Yang H, Liu W, Wang N, Yu X. Improved performance of magnetic cross-linked lipase aggregates by interfacial activation: A robust and magnetically recyclable biocatalyst for transesterification of jatropha oil. Molecules. 2017 Dec 1;22(12). Available from: <URL>.
  • 27. Sooch BS, Singh Kauldhar B. Influence of Multiple Bioprocess Parameters on Production of Lipase from Pseudomonas sp. BWS-5. Braz Arch Biol Technol. 2013;56(5):711–21. Available from: <URL>.
  • 28. Gusniah A, Veny H, Hamzah F. Activity and stability of immobilized lipase for utilization in transesterification of waste cooking oil. Bull Chem React Eng Catal. 2020 Apr 1;15(1):242–52. Available from: <URL>.
  • 29. Taskin M, Ucar MH, Unver Y, Kara AA, Ozdemir M, Ortucu S. Lipase production with free and immobilized cells of cold-adapted yeast Rhodotorula glutinis HL25. Biocatal Agric Biotechnol. 2016 Oct 1;8:97–103. Available from: <URL>.
  • 30. Turati DFM, Almeida AF, Terrone CC, Nascimento JMF, Terrasan CRF, Fernandez-Lorente G, et al. Thermotolerant lipase from Penicillium sp. section Gracilenta CBMAI 1583: Effect of carbon sources on enzyme production, biochemical properties of crude and purified enzyme and substrate specificity. Biocatal Agric Biotechnol. 2019;17:15–24. Available from: <URL>.
  • 31. Akhter K, Karim I, Aziz B, Bibi A, Khan J, Akhtar T. Optimization and characterization of alkaliphilic lipase from a novel Bacillus cereus NC7401 strain isolated from diesel fuel polluted soil. PLoS One. 2022 Aug 30;17(8):e0273368. Available from: <URL>.
  • 32. Emtenani S, Asoodeh A, Emtenani S. Molecular cloning of a thermo-alkaliphilic lipase from Bacillus subtilis DR8806: Expression and biochemical characterization. Process Biochem. 2013;48(11):1679–85. Available from: <URL>
  • 33. Salihu A, Alam MdZ. Solvent tolerant lipases: A review. Process Biochemistry [Internet]. 2015;50(1):86–96. Available from: <URL>.
  • 34. Hu J, Cai W, Wang C, Du X, Lin J, Cai J. Purification and characterization of alkaline lipase production by Pseudomonas aeruginosa HFE733 and application for biodegradation in food wastewater treatment. Biotechnol Biotechnol Equip. 2018 May 4;32(3):583–90. Available from: <URL>.
  • 35. Huang J, Yang Z, Zhu R, Qian X, Wang Y, Li Y, et al. Efficient heterologous expression of an alkaline lipase and its application in hydrolytic production of free astaxanthin. Biotechnol Biofuels. 2018;11(1):181. Available from: <URL>.
  • 36. Robinson PK. Enzymes: principles and biotechnological applications. Essays Biochem. 2015 Nov 15;59:1–41. Available from: <URL>.
  • 37. Sharma P, Sharma N, Pathania S, Handa S. Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry. J Genet Eng Biotechnol. 2017;15(2):369–77. Available from: <URL>.
  • 38. Bora L, Gohain D, Das R. Recent advances in production and biotechnological applications of thermostable and alkaline bacterial lipases. J Chem Technol Biotechnol. 2013 Nov 1;88(11):1959–70. Available from: <URL>.
  • 39. Kumar V, Yedavalli P, Gupta V, Rao NM. Engineering lipase A from mesophilic Bacillus subtilis for activity at low temperatures. Protein Eng Des Sel. 2014 Mar;27(3):73–82. Available from: <URL>.
  • 40. Fojan P, Jonson PH, Petersen MTN, Petersen SB. What distinguishes an esterase from a lipase: A novel structural approach. Biochimie. 2000;82:1033–41. Available from: <URL>.
There are 40 citations in total.

Details

Primary Language English
Subjects Biochemistry and Cell Biology (Other)
Journal Section RESEARCH ARTICLES
Authors

Azra Zahrah Nadhirah Ikhwani 0000-0002-8310-5013

Idris Idris 0000-0003-2565-5903

Rizki Rabeca Elfirta 0000-0001-6017-7236

Pamungkas Rizki Ferdian 0000-0002-4391-2902

Publication Date May 15, 2024
Submission Date July 28, 2023
Acceptance Date November 1, 2023
Published in Issue Year 2024

Cite

Vancouver Ikhwani AZN, Idris I, Elfirta RR, Ferdian PR. Production and Activity Characterization of Lipase from Bacillus flexus InaCC-B486. JOTCSA. 2024;11(2):397-404.