روان سازی و بازیافت نفت خام توسط بیوسورفاکتانت تولید شده از اسینتوباکتر جانسونی ABR6
محورهای موضوعی : زیست فناوری میکروبیالهام اکبری 1 , کیوان بهشتی مآل 2 , بهنام راسخ 3 , زرین دخت امامی کرونی 4 , میثم امیدی 5
1 - گروه میکروبیولوژی، دانشکده علوم زیستی، دانشگاه آزاد اسلامی واحد فلاورجان، اصفهان، ایران
2 - دانشیار، گروه میکروبیولوژی، دانشکده علوم زیستی، واحد فلاورجان، دانشگاه آزاد اسلامی، اصفهان، ایران
3 - گروه پژوهش میکروبیولوژی و بیوتکنولوژی، پژوهشگاه صنعت نفت، تهران، ایران
4 - دانشگاه آزاد اسلامی واحد فلاورجان، اصفهان، ایران.
5 - گروه مهندسی بافت، دانشکده فناوری های نوین پزشکی، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران
کلید واژه: بیوسورفاکتانت, روان سازی نفت خام, اسینتوباکتر جانسونی,
چکیده مقاله :
سابقه و هدف: پسماند های جامد و نیمه جامد به عنوان لجن نفتی در مراحل مختلف پالایش نفت خام تولید می شود. تجمع پسماند نفتی در پالایشگاه منجر به کاهش کارایی پالایش نفت و رهاسازی آن موجب آلودگی های زیست محیطی می شود. هدف از انجام این تحقیق روان سازی نفت خام در خطوط لوله و بازیافت نفت با استفاده از بیوسورفاکتانت تولید شده توسط یک سویه ی بومی می باشد.مواد و روش ها: جدایه های مولد بیوسورفاکتانت از مخازن نفت پالایشگاه اصفهان جداسازی شدند و غربال گری آن توسط آزمون گسترش نفت انجام شد. همچنین کشش سطحی با استفاده دستگاه تنسیومتر اندازه گیری شد. ساختار شیمیایی بیوسورفاکتانت با استفاده از آنالیزهای شیمیایی شناسایی شد. بازیافت نفت از لجن نفتی در شرایط ثابت اندازه گیری شد. تاثیرروان سازی بیوسور فاکتانت بر روی نفت خام در خطوط لوله در شرایط آزمایشگاهی ارزیابی گردید. همچنین پایداری بیوسورفاکتانت در شرایط محیطی مختلف اندازه گیری شد.یافته ها: بهترین باکتری تولید کننده ی بیوسورفاکتانت به عنوان اسینتوباکتر جانسونی ABR6شناسایی شد و توالی ژنومی 16S rDNA آن در بانک ژن با شماره ی دسترسی 100470 MK ثبت شد. آنالیز های شیمیایی اثبات کرد که بیوسورفاکتانت تولید شده لیپوپپتیدی است. به وسیله ی بیوسورفاکتانت حاصل از این باکتری 50 درصد نفت خام از لجن نفتی بازیافت شد و همچنین سرعت انتقال نفت خام در خطوط لوله از64 ثانیه به 35 ثانیه کاهش یافت. همچنین این بیوسورفاکتانت پایداری بالایی درw/v 5% نمک ، گسترهpH 8 تا 10 و دمای 60 درجه سلسیوس و شرایط اتوکلاو را داشت.نتیجه گیری: نتایج این پژوهش نشان داد که بیوسورفاکتانت تولید شده توسط سویه ی بومیاسینتوباکتر جانسونی ABR6علاوه بر فعالیت در شرایط افراطی، قابلیت بازیافت نفت و افزایش سرعت انتقال نفت خام در خطوط لوله را نیز دارد. بنابراین استفاده از بیوسورفاکتانت حاصل در صنایع نفت توصیه می شود.
Background and Objectives: Solid and semi-solid wastes are produced as oil sludge at different stages of crude oil refining. Accumulation of oil waste in the refinery reduces the efficiency of oil refining and its release causes environmental pollution. The purpose of this study is to lubricate crude oil in pipelines and recycle oil using a biosurfactant produced by an indigenous strain. Materials and Methods: The biosurfactant-producing isolates were obtained from petroleum reservoir in Isfahan Oil Refinery, Isfahan, Iran. Screening was performed by oil displacement method. Also the surface tension was measured by tensiometer. Biosurfactant chemical structure was identified by using chemical analysis. Oil recovery from the sludge was measured under controlled condition. The effect of biosurfactant lubrication was investigated on crude oil in the pipelines in vitro. The stability of biosurfactant in different environmental conditions was also determined. Results: The best biosurfactant-producing bacterium was identified as Acinetobacter johnsonii ABR6, and its 16S-rDNA genomic sequence was deposited in GenBank under the accession number of MK100470. Chemical analysis of TLC and FTIR confirmed that the produced biosurfactant was lipopeptide. The biosurfactant obtained from this bacterium recovered 50% of crude oil from petroleum sludge and also reduced transportation speed from 64 to 35 seconds. This biosurfactant had high stability in 5% w/v NaCl, pH range of 8 to 10, temperature of 60°C and autoclave conditions. Conclusion: The results of this study showed that the biosurfactant produced by the native strain of Acinetobacter Johnson ABR6, in addition to operating in extreme conditions, also has the ability to recover oil and increase the transfer rate of crude oil in pipelines. Therefore, the use of this biosurfactant can be considered as an asset in the petroleum industry
industry. Biodeg. 2012; 23,865–880.
2. Araújo S, Silva-Portela B, Lima, DC. A biosurfactant protein derived from a metagenomic
library with activity in oil degradation. Sci Rep. 2020; 10, 1340.
3. Bendaha M, Mebrek S, Naimi M. Isolation and comparison of rhamnolipids production in
Pseudomonas aeruginosa P.B:2 and Pseudomonas fluorescens P. . Inter Biodet
Biodeg,2012; 1–7444.
4. Banat IM and et al. Cost effective technologies and renewable substrates for biosurfactants'
production. fmicb, 2014; 5 697. 12
5. Banat IM, Franzetti A, Gandolfi I, Bestetti G and et al. Microbial biosurfactants production,
applications and future potential. Appl Microb Biotech.2010; 87:427-444.
6. Fenibo E, Ijimo G, Selvarjan R, Chiekere C, Microbial Surfactants: The Next Generation
Multifunctional Biomolecules for Applications in the Petroleum Industry and Its Associated
Environmental Remediation. Microb. 2019; 7(11): 581.
7. Ghorbannezhad H, Moghimi H, Dastgheib M. Biodegradation of high-molecular-weight
aliphatic and aromatic hydrocarbons by Aspergillus calidoustus, J of Micro.World.2018. 10:
4,346-359. [In Persian]
8. Alvarez M, Jurelevicius D, Marques JM, Souza PM and et al. Bacillus amyloliquefaciens
TSBSO 3.8, a biosurfactant-producing strain with biotechnological potential for microbial
enhanced oil recovery. Coll Surf B Biointerf.2015; 136:14-21.
9. EI-Sheshtawy HS, Doheim M Selection of Pseudomonas aeruginosa for biosurfactant
production and studies of its antimicrobial activity. Egyp J Petr.2014; 23:1-6..
10. El-Sheshtawy HS, Aiad I, Osman ME, Abo-ELnasr AA, Kobisy AS. Production of
biosurfactant from Bacillus licheniformis for microbial enhanced oil recovery and inhibition
the growth of sulfate reducing bacteria. Egyp J Petr.2015; 24:155–162.
11. Gudiña EJ, Fernandes EC, Rodrigues AI, Teixeira JA and et al. Biosurfactant production by
Bacillus subtilis using corn steep liquor as culture medium. Front Microb. 2015; 59:1-7..
12. He C, Dong W, Li J, Li Y, Huang C, Ma Y. Characterization of rhamnolipid biosurfactants
produced by recombinant Pseudomonas aeruginosa strain DAB with removal of crude oil.
Biotech Lett.2017; 39:1381-1388.
13. Akbari E, Beheshti-Maal K, Nayeri H. Production and optimization of alkaline lipase by a
novel psychrotolerant and halotolerant strain Planomicrobium okeanokoites ABN-IAUF-2
isolated from Persian Gulf. Inter Jl Med Res & Heal Scie.2016; 5:139–148.
14. Akbari E, Beheshti-Maal K, Nayeri H. A novel halo-alkalo-tolerant bacterium, Marinobacter
alkaliphilus ABN-IAUF-1, isolated from Persian Gulf suitable for alkaline lipase production.
IJEST.2018; 15:1767-1776.
15. Jafari A, Raheb J, Bardania H and et al. Isolation, cloning and expression of rhamnolipid
operon from Pseudomonas aeruginosa ATCC 9027 in logarithmic phase in E. coli BL21. Ame
J lif Scie.2015; 2:22-30.
16. Lai C-C, Huang Y-C, Wei Y-H, Chang J-S. Biosurfactant-enhanced removal of total
petroleum hydrocarbons from contaminated soil. J Hazard Mater. 2009;167:609–614.
17. Saikia RR, Deka S, Deka M, Banat IM. Isolation of biosurfactant producing Pseudomonas
aeruginosa RS29 from oil-contaminated soil and evaluation of different nitrogen sources in
biosurfactant production. Annal. Microb.2012; 62:753–763.
18. Lima TMS, Fonseca AF, Leão BA, Mounteer AH et al. Oil recovery from fuel oil storage tank
sludge using biosurfactants. J Bio. Biodeg. 2011; 2:1-5.
19. Amani H and Kariminezhad H. Study on emulsification of crude oil in water using emulsan
biosurfactant for pipeline transportation. Pet. Sci. Tech. 2016; 34,216–222
20. Mouafi F, Elsoud M, et al Optimization of biosurfactant production by Bacillus brevis using
response surface methodology. biotech. Report.2016; 7: 1092
21. Khademolhosseini R, Jafari A, Mosavi S, and et al. Physicochemical characterization and
optimization of glycolipid biosurfactant production by a native strain of Pseudomonas
aeruginosa HAK01 and its performance evaluation for the MEOR process. RSC Adv., 2019;
9, 7932-7947
22. Deepika, K. ., B.A. Kumar, S. Gnanender and P. . Bramhachari, Pseudomonas aeruginosa
K D1 an efficient biosurfactant producing bacteria isolated from Krishna Delta Mangrove
Sediments. Res. J. Environ. Sci.,2014, 8: 134-141
23. Ghorbannezhad H, Moghimi H, Dastgheib M. Biodegradation of high-molecular-weight
aliphatic and aromatic hydrocarbons by Aspergillus calidoustus, J of Micro.World.2018; 10:
4,346-359.[ In Persian]
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industry. Biodeg. 2012; 23,865–880.
2. Araújo S, Silva-Portela B, Lima, DC. A biosurfactant protein derived from a metagenomic
library with activity in oil degradation. Sci Rep. 2020; 10, 1340.
3. Bendaha M, Mebrek S, Naimi M. Isolation and comparison of rhamnolipids production in
Pseudomonas aeruginosa P.B:2 and Pseudomonas fluorescens P. . Inter Biodet
Biodeg,2012; 1–7444.
4. Banat IM and et al. Cost effective technologies and renewable substrates for biosurfactants'
production. fmicb, 2014; 5 697. 12
5. Banat IM, Franzetti A, Gandolfi I, Bestetti G and et al. Microbial biosurfactants production,
applications and future potential. Appl Microb Biotech.2010; 87:427-444.
6. Fenibo E, Ijimo G, Selvarjan R, Chiekere C, Microbial Surfactants: The Next Generation
Multifunctional Biomolecules for Applications in the Petroleum Industry and Its Associated
Environmental Remediation. Microb. 2019; 7(11): 581.
7. Ghorbannezhad H, Moghimi H, Dastgheib M. Biodegradation of high-molecular-weight
aliphatic and aromatic hydrocarbons by Aspergillus calidoustus, J of Micro.World.2018. 10:
4,346-359. [In Persian]
8. Alvarez M, Jurelevicius D, Marques JM, Souza PM and et al. Bacillus amyloliquefaciens
TSBSO 3.8, a biosurfactant-producing strain with biotechnological potential for microbial
enhanced oil recovery. Coll Surf B Biointerf.2015; 136:14-21.
9. EI-Sheshtawy HS, Doheim M Selection of Pseudomonas aeruginosa for biosurfactant
production and studies of its antimicrobial activity. Egyp J Petr.2014; 23:1-6..
10. El-Sheshtawy HS, Aiad I, Osman ME, Abo-ELnasr AA, Kobisy AS. Production of
biosurfactant from Bacillus licheniformis for microbial enhanced oil recovery and inhibition
the growth of sulfate reducing bacteria. Egyp J Petr.2015; 24:155–162.
11. Gudiña EJ, Fernandes EC, Rodrigues AI, Teixeira JA and et al. Biosurfactant production by
Bacillus subtilis using corn steep liquor as culture medium. Front Microb. 2015; 59:1-7..
12. He C, Dong W, Li J, Li Y, Huang C, Ma Y. Characterization of rhamnolipid biosurfactants
produced by recombinant Pseudomonas aeruginosa strain DAB with removal of crude oil.
Biotech Lett.2017; 39:1381-1388.
13. Akbari E, Beheshti-Maal K, Nayeri H. Production and optimization of alkaline lipase by a
novel psychrotolerant and halotolerant strain Planomicrobium okeanokoites ABN-IAUF-2
isolated from Persian Gulf. Inter Jl Med Res & Heal Scie.2016; 5:139–148.
14. Akbari E, Beheshti-Maal K, Nayeri H. A novel halo-alkalo-tolerant bacterium, Marinobacter
alkaliphilus ABN-IAUF-1, isolated from Persian Gulf suitable for alkaline lipase production.
IJEST.2018; 15:1767-1776.
15. Jafari A, Raheb J, Bardania H and et al. Isolation, cloning and expression of rhamnolipid
operon from Pseudomonas aeruginosa ATCC 9027 in logarithmic phase in E. coli BL21. Ame
J lif Scie.2015; 2:22-30.
16. Lai C-C, Huang Y-C, Wei Y-H, Chang J-S. Biosurfactant-enhanced removal of total
petroleum hydrocarbons from contaminated soil. J Hazard Mater. 2009;167:609–614.
17. Saikia RR, Deka S, Deka M, Banat IM. Isolation of biosurfactant producing Pseudomonas
aeruginosa RS29 from oil-contaminated soil and evaluation of different nitrogen sources in
biosurfactant production. Annal. Microb.2012; 62:753–763.
18. Lima TMS, Fonseca AF, Leão BA, Mounteer AH et al. Oil recovery from fuel oil storage tank
sludge using biosurfactants. J Bio. Biodeg. 2011; 2:1-5.
19. Amani H and Kariminezhad H. Study on emulsification of crude oil in water using emulsan
biosurfactant for pipeline transportation. Pet. Sci. Tech. 2016; 34,216–222
20. Mouafi F, Elsoud M, et al Optimization of biosurfactant production by Bacillus brevis using
response surface methodology. biotech. Report.2016; 7: 1092
21. Khademolhosseini R, Jafari A, Mosavi S, and et al. Physicochemical characterization and
optimization of glycolipid biosurfactant production by a native strain of Pseudomonas
aeruginosa HAK01 and its performance evaluation for the MEOR process. RSC Adv., 2019;
9, 7932-7947
22. Deepika, K. ., B.A. Kumar, S. Gnanender and P. . Bramhachari, Pseudomonas aeruginosa
K D1 an efficient biosurfactant producing bacteria isolated from Krishna Delta Mangrove
Sediments. Res. J. Environ. Sci.,2014, 8: 134-141
23. Ghorbannezhad H, Moghimi H, Dastgheib M. Biodegradation of high-molecular-weight
aliphatic and aromatic hydrocarbons by Aspergillus calidoustus, J of Micro.World.2018; 10:
4,346-359.[ In Persian]