بیوسنتز نانوذره مس توسط باکتری استنوتروفوموناس مالتوفیلیا
محورهای موضوعی : زیست فناوری میکروبی
زهرا امیرپور
1
,
منیر دودی
2
,
غلام رضا امیری
3
1 - کارشناس ارشد، گروه میکروب شناسی، واحد فلاورجان، دانشگاه آزاد اسلامی، اصفهان، ایران
2 - استادیار، گروه میکروب شناسی، واحد فلاورجان، دانشگاه آزاد اسلامی، اصفهان، ایران
3 - استادیار، گروه زیست شناسی، واحد فلاورجان، دانشگاه آزاد اسلامی، اصفهان، ایران
کلید واژه: XRD, UV-Vis, بیوسنتز, نانوذره مس, .TEM,
چکیده مقاله :
سابقه و هدف: فلزات سنگین از آلاینده های پایدار و با دوام محیط زیست هستند که به یک مشکل جهانی تبدیل شده اند. با توجه به این که میکروارگانیسم ها مقاومت بالایی نسبت به فلزات دارند و موجب پاک سازی محیط زیست و تولید نانو ذرات می شوند، پژوهش حاضر به منظور تولید نانو ذره مس از باکتری های مقاوم به این فلز از پساب دو کارگاه مسگری در اصفهان برنامه ریزی شد.مواد و روش ها: از پساب دو کارگاه مسگری در شهر اصفهان به صورت مقطعی نمونه برداری شد. عوامل فیزیکوشیمیایی پساب ها، حداقل غلظت ممانعت کننده از رشد باکتری ها (MIC) به مس و مقاومت آنها نسبت به چند آنتی بیوتیک بررسی گردید. در ادامه آزمون های شناسایی مورفولوژی، بیوشیمیایی و مولکولی بر روی نمونه ها انجام شد. سپس به منظورارزیابی ساخت نانوذرات مس، بیومس باکتری مقاوم به مس به محلول ذخیره سولفات مس افزوده شد و نتایج توسط دستگاه طیف سنج فرابنفش- مرئی (UV-VIS)، تفرق اشعه ایکس (XRD) و میکروسکوپ الکترونی گذاره (TEM) مورد ارزیابی قرار گرفت.یافته ها: از میان باکتری های مورد بررسی، باکتری باسیلوس تویوننسیس سویه NE2 با mM 3/5 MIC=و آرتروباکتر آژیلیس سویه NE1 با mM 4 MIC=از پساب مسگری شماره 2 و باکتری استنوتروفوموناس مالتوفیلیا سویه 5633 با mM 6 MIC=از پساب مسگری شماره 1 جداسازی شدند. از این میان تنها باکتری استنوتروفوموناس مالتوفیلیا سویه 5633 قادر به سنتز نانوذرات مس بود. پیک های ایجاد شده در محدوده ۴۳۰-۲۵۰ نانومتر، تایید کننده ذرات مس (Cu) و اکسید مس (CuO) بودند.نتیجه گیری: یافته های این پژوهش نشان داد که باکتری جداسازی شده می تواند کاندید مناسبی به منظور حذف مس از پساب ها و همچنین تولید کننده زیستی نانو ذره مس باشد.
Background & Objectives: Heavy metals are environmentally sustainable and durable pollutants that have become a world problem. As microorganisms show high resistance to heavy metals and can purify the environment and produce nanoparticles, the present study was designed to produce copper nanoparticles from copper-resistant bacteria isolated from wastewater of two copper workshops in Isfahan. Materials & Methods: In this cross-sectional study, wastewater samples were collected from two copper workshops in Isfahan. The physicochemical factors of the wastewater, the minimum inhibitory concentration of bacteria (MIC) to copper and their resistance to several antibiotics were investigated. Morphological, biochemical and molecular identification tests were carried out on samples. Then the biomass of copper-resistant bacteria was added to the copper sulfate pentahydrate stock (CuSO4.5H2O) and the results were evaluated by Ultraviolet-Visible spectrophotometer (UV-VIS), X-ray diffraction (XRD) and Transient Electron Microscopy (TEM). Results: Among the studied bacteria, the Bacillus toyonensis strain NE2 with the MIC of 3.5 mM and Arthrobacteragilis NE1 with MIC of 4 mM from copper workshop 2 and Stenotrophomonas maltophilia strain 5633 with the MIC of 6 mM from copper workshop 1 were isolated. Among these isolates, only S. maltophilia strain 5633 was able to synthesize copper nanoparticles. Peaks created in the range of 250-430 nm confirmed the presence of Copper (Cu) and Copper Oxide (CuO) particles. Conclusion: The findings of this study showed that the isolated bacteria could be a good candidate to remove copper from wastewater and to biosynthesize copper nanoparticle.
Bio Med. 2010; 6(2): 257-262.
2. Mandal D, Bolander ME, Mukhopadhyay D, Sarkar G, Mukherjee P. The use of
microorganisms for the formation of metal nanoparticles and their application. J Appl
Microbiol Biotech. 2006; 69(5): 485-492.
3. Mohaseli T, Pourseyedi SH. Synthesis green and silver nanoparticle characterization using
aqueous extracts of sesame plant. J Biotech Tarbiat Modarres Uni. 2012; 6(1): 10-20.
[In Persian]
4. Emtiazi G. 2013. Investigation of the effective genes in the resistance of heavy metals and steel
nano oxides in isolated Pseudomonads and their biological control by microbial antagonists.
Faculty Basic Sciences Isfahan University.
5. Pourali P, Baseri Salehi M, Afsharnezhad S, Behravan J. Biological production and assessment
of the antibacterial activity of gold nanoapraticles. J Microbial World. 2013; 6(3): 198-211.
[In Persian]
6. Hosseini MR , Shaffie M. Kinetic study of the biological copper removal from a copper
sulfate solution in the form of copper sulfide nanoparticles. J Sep Sci Eng. 2013; 4(2): 75-82.
[In Persian]
7. Suleiman M, Mousa M, Hussein A. Wastewater disinfection by synthesized copper oxide
nanoparticles stabilized with surfactant. J Mat Env Sci. 2015; 6(7): 1924-1937.
8. Mirhendi M. 2012. Production of iron, zinc and copper nanoparticles by microbial biofilm and
pure culture. Faculty Basic Sciences Isfahan University.
9. Abboud Y, Saffaj T, Chagraoui A, El Bouari A, Brouzi K. Biosynthesis, characterization and
antimicrobial activity of copper oxide nanoparticles (CONPs) produced using brown alga
extract (Bifurcaria bifurcata). J Appl Nanosci. 2014; 4: 571-576.
10. Grass G, Stanto CE, Morais PV. Isolation and characterization of bacteria resistant to metallic
copper surfaces. Appl Environ Microbiol. 2010; 5: 134-1348.
11. Shahsanaie Ganirani M, Ahadi AM, Doudi M. Biosorption of silver by bacteria
Stenoterophomonas maltophilia strains MS8 from the wastewater treatment plant in Isfahan
silver. J Microbial World. 2015; 8(2): 158-167. [In Persian]
12. Alboghobeysh H, Tahmourespour A, Doudi M. Antibiotic resistance in isolated bacteria from
urban sewage and copper smeltery industrial wastewater. J Gorgan Uni Med Sci .2013; 15(1):
95-102. [In Persian]
13. Rajbanshi A. Study on heavy metal resistant bacteria in Guheswori Sewage treatment plant. J
Our Nature. 2008; 6(1): 52-57.
14. Ahrabi M, Hosseini F, Akhavan Sepahi A. Isolation and identification of bacteria with
potential environmental cleanup selenite salts of environmental resources. J Biotech Cell
Molecule Bio. 2013; 4(13): 86-91. [In Persian]
15. Rahman A, Ismail A, Jumbianti D, Maghalena S, Sudrajat H. Synthesis of copper oxid nano
particles by using Phormidium cyanobacterium. Indonesia J Chem. 2009; 9(3): 355-360.
16. Vashney R, Bhadauria S, Gaur MS, Pasricha R. Characterization of copper nanoparticles
synthesized by a novel microbiological method. J Min Met Mat Society. 2010; 62(12):
102-104.
17. Qrayelu D, Moein darbari M. Factors contributing to the spread of appropriate samples for
laboratory imaging TEM. Iran J Lab Know. 2013; 1: 30-36. [In Persian]
18. Amiri GHR, Yousefi MH, Abolhassani MR, Manouchehri S, Keshavarz MH, Fatahian S.
Magnetic properties and microwave absorption in Ni–Zn and Mn–Zn ferrite nanoparticles
synthesized by low-temperature solid-state reaction. J Mag Mag Mater. 2010; 323(6): 730-734.
19. Laila MA, Wagdy KB, Thanaa HA, Karima FM. Heavy metal resistance and gene expression
analysis of metal resistance gene in Gram-negative and Gram-positive bacteria present in
Egyption soil. J Appl Sci Env Sanit. 2011; 6(2): 201-211.
20. Edward raja C, Selvam GS, Omine K. Isolation, identification and characterization of heavy
metal resistant bacteria from sewage. Int J Sym Geo disaster prevent Geo Asia. 2009; 205-211.
21. Sengupta S, Chattopadhyay MK. Antibiotic resistance of bacteria: A global challenge. J Sci Ed.
2012; 17(2): 177-191.
22. Seeger M, Altimira F, Bravo G, Gonzalez M, Rojas LA, Yanez C. Characterization of
copper-resistant bacteria and bacterial communities from copper-polluted agricultural soils of
central Chile. J Bio Med Cent Microbiol. 2012; 12(193): 1-12.
23. Grass G, Morais PV, Santo CE. Isolation and characterization of bacteria resistant to metallic
Copper surfaces. J Appl Env Microbiol. 2010; 77(5): 1341-1348.
24. Virender S, Chauhan PK, Kanta R, Dhewa T. Isolation and characterization of Pseudomonas
resistant to heavy metal contaminants. Int J Pharmacol Sci Rev Res. 2010; 3(2): 164-167.
25. Egbebi AO, Famurewa O. Heavy metal resistance among Klebsiella isolates in some parts of
southwest Nigeria. Asia J Med Sci. 2011; 3(5): 183-185.
26. Varenyam A, Qiuzhuo Z, Wei-Ning X, Duanchao W. Identification of heavy metal resistant
bacteria isolated from Yangtz River, China. Int J Agri Bio. 2014; 16(3): 619-623.
27. Nasr Azadani A, Tahmourespour A, Hoodaji M. Determination of the threshold of bacterial
tolerance to lead, zinc and cadmium in three types of industrial wastewater. J Ecol. 2010; 36
(56): 75-86. [In Persian]
28. Anyanwu CU, Nwachukwu ON. Heavy metal resistance in bacteria isolated from contaminated
and uncontaminated soils. Int J Res Chem En. 2011; 1(1): 173-178.
29. Wang G, Shi Z, Cao Z, Qin D, Zhu W, Wang Q, Li M. Correlation models between
environmental factors and bacterial resistance to antimony and copper. Plos One. 2013;
8(10): 1-11.
30. Akhavan SA, Sharifian S, Zolfaghari MR, Khalili DM. Evaluation of resistance in intestinal
coli forms isolated from industrial wastewater, domestic wastewater and various parts of the
wastewater treatment system of Arak city. J Cell Mol Res. 2014; 27(2): 167-178. [In Persian]
31. Zayats AV, Smolyaninov I, Maradudin A. Nano-optics of surface plasmon polaritons. J Phy
Rep. 2005; 408(3): 131-314.
32. Acharyulu NPS, Dubey RS, Swaminadham V, Kalyani RL, Kollu P, Pammi SVN. Green
synthesis of CuO nanoparticles using Phyllanthus amarus leaf extract and their antibacterial
activity against multidrug resistance bacteria. Int J Eng Res Tech. 2014; 4(3): 639-641.
33. Shaffiey SF, Ahmadi M, Shaffiey SR, Shapoori M, Varshoie H, Azari F. Synthesis of copper
oxide (CuO) nanoparticles and surveying its bactericidal properties against Aeromonas
hydrophila bacteria. J Fasa Uni Med Sci. 2014; 5(1): 36-43. [In Persian]
34. Eftekhari BS. The physical properties of nano-systems approach in the design of drug
delivery. J Month Nanotech. 2016; 15(3): 32-35. [In
_||_
Bio Med. 2010; 6(2): 257-262.
2. Mandal D, Bolander ME, Mukhopadhyay D, Sarkar G, Mukherjee P. The use of
microorganisms for the formation of metal nanoparticles and their application. J Appl
Microbiol Biotech. 2006; 69(5): 485-492.
3. Mohaseli T, Pourseyedi SH. Synthesis green and silver nanoparticle characterization using
aqueous extracts of sesame plant. J Biotech Tarbiat Modarres Uni. 2012; 6(1): 10-20.
[In Persian]
4. Emtiazi G. 2013. Investigation of the effective genes in the resistance of heavy metals and steel
nano oxides in isolated Pseudomonads and their biological control by microbial antagonists.
Faculty Basic Sciences Isfahan University.
5. Pourali P, Baseri Salehi M, Afsharnezhad S, Behravan J. Biological production and assessment
of the antibacterial activity of gold nanoapraticles. J Microbial World. 2013; 6(3): 198-211.
[In Persian]
6. Hosseini MR , Shaffie M. Kinetic study of the biological copper removal from a copper
sulfate solution in the form of copper sulfide nanoparticles. J Sep Sci Eng. 2013; 4(2): 75-82.
[In Persian]
7. Suleiman M, Mousa M, Hussein A. Wastewater disinfection by synthesized copper oxide
nanoparticles stabilized with surfactant. J Mat Env Sci. 2015; 6(7): 1924-1937.
8. Mirhendi M. 2012. Production of iron, zinc and copper nanoparticles by microbial biofilm and
pure culture. Faculty Basic Sciences Isfahan University.
9. Abboud Y, Saffaj T, Chagraoui A, El Bouari A, Brouzi K. Biosynthesis, characterization and
antimicrobial activity of copper oxide nanoparticles (CONPs) produced using brown alga
extract (Bifurcaria bifurcata). J Appl Nanosci. 2014; 4: 571-576.
10. Grass G, Stanto CE, Morais PV. Isolation and characterization of bacteria resistant to metallic
copper surfaces. Appl Environ Microbiol. 2010; 5: 134-1348.
11. Shahsanaie Ganirani M, Ahadi AM, Doudi M. Biosorption of silver by bacteria
Stenoterophomonas maltophilia strains MS8 from the wastewater treatment plant in Isfahan
silver. J Microbial World. 2015; 8(2): 158-167. [In Persian]
12. Alboghobeysh H, Tahmourespour A, Doudi M. Antibiotic resistance in isolated bacteria from
urban sewage and copper smeltery industrial wastewater. J Gorgan Uni Med Sci .2013; 15(1):
95-102. [In Persian]
13. Rajbanshi A. Study on heavy metal resistant bacteria in Guheswori Sewage treatment plant. J
Our Nature. 2008; 6(1): 52-57.
14. Ahrabi M, Hosseini F, Akhavan Sepahi A. Isolation and identification of bacteria with
potential environmental cleanup selenite salts of environmental resources. J Biotech Cell
Molecule Bio. 2013; 4(13): 86-91. [In Persian]
15. Rahman A, Ismail A, Jumbianti D, Maghalena S, Sudrajat H. Synthesis of copper oxid nano
particles by using Phormidium cyanobacterium. Indonesia J Chem. 2009; 9(3): 355-360.
16. Vashney R, Bhadauria S, Gaur MS, Pasricha R. Characterization of copper nanoparticles
synthesized by a novel microbiological method. J Min Met Mat Society. 2010; 62(12):
102-104.
17. Qrayelu D, Moein darbari M. Factors contributing to the spread of appropriate samples for
laboratory imaging TEM. Iran J Lab Know. 2013; 1: 30-36. [In Persian]
18. Amiri GHR, Yousefi MH, Abolhassani MR, Manouchehri S, Keshavarz MH, Fatahian S.
Magnetic properties and microwave absorption in Ni–Zn and Mn–Zn ferrite nanoparticles
synthesized by low-temperature solid-state reaction. J Mag Mag Mater. 2010; 323(6): 730-734.
19. Laila MA, Wagdy KB, Thanaa HA, Karima FM. Heavy metal resistance and gene expression
analysis of metal resistance gene in Gram-negative and Gram-positive bacteria present in
Egyption soil. J Appl Sci Env Sanit. 2011; 6(2): 201-211.
20. Edward raja C, Selvam GS, Omine K. Isolation, identification and characterization of heavy
metal resistant bacteria from sewage. Int J Sym Geo disaster prevent Geo Asia. 2009; 205-211.
21. Sengupta S, Chattopadhyay MK. Antibiotic resistance of bacteria: A global challenge. J Sci Ed.
2012; 17(2): 177-191.
22. Seeger M, Altimira F, Bravo G, Gonzalez M, Rojas LA, Yanez C. Characterization of
copper-resistant bacteria and bacterial communities from copper-polluted agricultural soils of
central Chile. J Bio Med Cent Microbiol. 2012; 12(193): 1-12.
23. Grass G, Morais PV, Santo CE. Isolation and characterization of bacteria resistant to metallic
Copper surfaces. J Appl Env Microbiol. 2010; 77(5): 1341-1348.
24. Virender S, Chauhan PK, Kanta R, Dhewa T. Isolation and characterization of Pseudomonas
resistant to heavy metal contaminants. Int J Pharmacol Sci Rev Res. 2010; 3(2): 164-167.
25. Egbebi AO, Famurewa O. Heavy metal resistance among Klebsiella isolates in some parts of
southwest Nigeria. Asia J Med Sci. 2011; 3(5): 183-185.
26. Varenyam A, Qiuzhuo Z, Wei-Ning X, Duanchao W. Identification of heavy metal resistant
bacteria isolated from Yangtz River, China. Int J Agri Bio. 2014; 16(3): 619-623.
27. Nasr Azadani A, Tahmourespour A, Hoodaji M. Determination of the threshold of bacterial
tolerance to lead, zinc and cadmium in three types of industrial wastewater. J Ecol. 2010; 36
(56): 75-86. [In Persian]
28. Anyanwu CU, Nwachukwu ON. Heavy metal resistance in bacteria isolated from contaminated
and uncontaminated soils. Int J Res Chem En. 2011; 1(1): 173-178.
29. Wang G, Shi Z, Cao Z, Qin D, Zhu W, Wang Q, Li M. Correlation models between
environmental factors and bacterial resistance to antimony and copper. Plos One. 2013;
8(10): 1-11.
30. Akhavan SA, Sharifian S, Zolfaghari MR, Khalili DM. Evaluation of resistance in intestinal
coli forms isolated from industrial wastewater, domestic wastewater and various parts of the
wastewater treatment system of Arak city. J Cell Mol Res. 2014; 27(2): 167-178. [In Persian]
31. Zayats AV, Smolyaninov I, Maradudin A. Nano-optics of surface plasmon polaritons. J Phy
Rep. 2005; 408(3): 131-314.
32. Acharyulu NPS, Dubey RS, Swaminadham V, Kalyani RL, Kollu P, Pammi SVN. Green
synthesis of CuO nanoparticles using Phyllanthus amarus leaf extract and their antibacterial
activity against multidrug resistance bacteria. Int J Eng Res Tech. 2014; 4(3): 639-641.
33. Shaffiey SF, Ahmadi M, Shaffiey SR, Shapoori M, Varshoie H, Azari F. Synthesis of copper
oxide (CuO) nanoparticles and surveying its bactericidal properties against Aeromonas
hydrophila bacteria. J Fasa Uni Med Sci. 2014; 5(1): 36-43. [In Persian]
34. Eftekhari BS. The physical properties of nano-systems approach in the design of drug
delivery. J Month Nanotech. 2016; 15(3): 32-35. [In