بهینه سازی تولید پلی هیدروکسی‌بوتیرات سویه جدید باسیلوس سرئوس جدا شده از پساب پتروشیمی
محورهای موضوعی : میکروب شناسی صنعتی
خسرو عیسی‌زاده
1
(دانشگاه آزاد اسلامی واحد لاهیجان، دانشکده ی علوم پایه، گروه میکروبیولوژی)
صبا امیری کجوری
2
(گروه میکروب شناسی، دانشکده علوم پایه، واحد لاهیجان، دانشگاه آزاد اسلامی، لاهیجان، ایران.)
زهیر حشمتی پور
3
(گروه میکروب شناسی، واحد تنکابن، دانشگاه آزاد اسلامی، تنکابن، ایران.)
میرساسان میرپور
4
(گروه میکروب شناسی، دانشکده علوم پایه، واحد لاهیجان، دانشگاه آزاد اسلامی، لاهیجان، ایران.)
سعید ضرابی
5
(گروه شیمی، دانشکده علوم پایه، واحد لاهیجان، دانشگاه آزاد اسلامی، لاهیجان، ایران.)
کلید واژه: بهینه سازی, پلی هیدروکسی‌بوتیرات, پساب پتروشیمی, پلاستیک زیستی,
چکیده مقاله :
سابقه و هدف: پلی هیدروکسی بوتیرات یک بیوپلی مر تولید شده توسط باکتری ها است. این ترکیبات با وجود داشتن خواص فیزیکی و شیمیایی مشابه با پلاستیک های مصنوعی کاملاً زیست تخریب پذیر و سازگار با محیط زیست می باشند. این مطالعه با هدف جداسازی و شناسایی یک سویه بومی تولید کننده پلی هیدروکسی بوتیرات با قابلیت تولید مناسب برای اهداف صنعتی انجام شد. مواد و روش ها: در این مطالعه تحلیلی، نمونه برداری از پساب شرکت پتروشیمی انجام شد. وجود پلی هیدروکسی بوتیرات در جدایه ها با رنگ آمیزی سودان سیاه مورد بررسی قرار گرفت. یک جدایه باسیلوس به منظور افزایش تولید پلی هیدروکسی بوتیرات انتخاب شد. این جدایه با روش های بیوشیمیایی و تعیین توالی ژن 16S rRNA شناسایی شد.تأیید نهایی سنتز پلی هیدروکسی بوتیرات توسط آنالیزهای طیف سنجی مادون قرمز و رزونانس مغناطیسی هسته پروتون انجام شد. به منظور تولید بیشتر پلی هیدروکسی بوتیرات تأثیر عوامل مختلف مانند منابع کربن و نیتروژن، pH و دما ارزیابی شدند.یافته ها: در مجموع 8 جدایه باکتریایی تولید کننده پلی هیدروکسی بوتیرات جداسازی شد که از این میان یک سویه جدید باسیلوس سرئوس به عنوان جدایه برتر انتخاب گردید. شرایط بهینه به منظور تولید بیشتر پلی هیدروکسی بوتیرات، استفاده از گلوکز به عنوان منبع کربن، سولفات آمونیوم به عنوان منبع نیتروژن، pH 7 و دمای 30 درجه سلسیوس بود.نتیجه گیری: مطالعه حاضر نتایج ارزشمندی را در مورد شرایط بهینه برای تولید پلی هیدروکسی بوتیرات فراهم آورده است که می تواند در سطح صنعتی برای تولید پلی هیدروکسی بوتیرات به عنوان یک جایگزین سریع پلاستیک های مصنوعی مبتنی بر نفت مورداستفاده قرار گیرد.
Background & Objectives: Polyhydroxybutyrate is a biopolymer Produced by bacteria. Also, with having chemical and physical characteristics similar with artificial plastics are completely biodegradable and compatible with life-environment. This study has been done with the purpose of separating and knowing one local strain with the ability of high-production for industrial purposes.Materials & Methods: In the present analytical research, the sampling from petrochemical wastewater has been done. The existence of polyhydroxybutyrate in separations has been studied with Sudan black staining. One separation of bacillus was chosen to increase the production of polyhydroxybutyrate. This separation was distinguished with biochemical methods and determining 16S rRNA gene sequencing. The final confirmation of Polyhydroxybutyrate synthesis was done through Fourier transform infrared spectroscopy and Proton nuclear magnetic resonance. To increase more production of polyhydroxybutyrate, the effect of different factors including carbone, nitrogen, pH and temperature were assessed. Results: As a whole, eight bacterial isolation producing polyhydroxybutyrate were separated that among them the one novel strain of bacillus cereus was chosen as better separation. The best conditions to increase more production of polyhydroxybutyrate, was the application and use of glucose as carbon source, ammonium sulphate as nitrogen source, 7 pH and 30°C temperature.Conclusion: Valuable data on optimized conditions for PHB production has been obtained through the present research that can be used at industrial level for PHB production, so it can be called a quick appearing alternation for petroleum based on synthetic plastic.
362- 367.
2. Shivalkar Y , Prabha R. Polyhydroxybutyrate as bio-degradable plastic – a review. J Environ
Sci Tox Food Technol. 2017, 11 (5): 10-12.
3. Sharma M, Dhingra H . Poly-β-hydroxybutyrate: a biodegradable polymer, biosynthesis and
biodegradation. BMJR. 2016; 14(3): 1-11.
4. Vijaya C, Reddy RM. Impact of soil composting using municipal solid waste on
biodegradation of plastics. Indian J Biotechnol. 2008; 7: 23 –239.
5. Reddy RL, Reddy VS, upta A. Study of bio-plastics as green and sustainable alternative to
plastics. Int J of Emerging Technol and Advanced Eng. 2013; 3(5): 82-89.
6. etachew A, Woldesenbet F. Production of biodegradable plastic by polyhydroxybutyrate
(PHB) accumulating bacteria using low cost agricultural waste material. BMC Res
Notes. 2016; 9(509): 1-9.
7. Al- addo HB, Sudesh , Samian MR. Screening of bacteria for PHA production using
waste glycerol as carbon source and the ability of new strain to produce P(3HB-co-3HV)
copolymer. Mal J Microbiol. 2016; 12(3): 245-253.
8. Panchal Sh. Recent developments on biodegradable polymers and their future trends. Int Res J
Sci Eng. 2016; 4 (1): 17-26.
9. Sharma M, Dhingra H . Poly-β-hydroxybutyrate: a biodegradable polyester, biosynthesis and
biodegradation. British Microbiology Res J. 2016; 14(3): 1-11.
10. Chaitanya , Mahmood S , ausar R, Sunilkumar N. Biotechnological production of
polyhydroxyalkanoates by various isolates: a review. Int J Pharma Sci Invent. 2014; 3(9):
111.
11. UMA Maheswari M, Dhandayuthapani . Fermentation optimization for production of
polyhydroxybutyrate (PHB) by newly isolated Azotobacter vinelandii DP. Int J Pharm
BioSci. 2013; 4(3): 779-787.
12. Shah R. Optimization and production of polyhydroxybutyrate (PHB) by Bacillus subtilis
1S1 from soil. Int J Curr Microbiol App Sci. 2014; 3(5): 377-387.
13. Jangra MR, Nehra S. Isolation, screening and characterization of new strains with
optimization studies to augment bacterial PHB production. Bull Env Pharmacol Life Sci.
2017; 6(8): 34-44.
14. Nehra , Chhabra N, Sidhu P , Lathwal P, Rana JS. Molecular identification and
characterization of poly-β-hydroxybutyrate (PHB) producing bacteria isolated from
contaminated soils. Asian J Microbiol Biotech Env Sci. 2015; 17(4): 281-290.
15. Bhagowati P, Pradhan Sh, Hirak R, Surajit D. Production, optimization and characterization
of polyhydroxybutyrate, a biodegradable plastic by Bacillus spp. Biosci Biotech
Biochem. 2015; 79(9): 1454- 1463.
16. Nehra , Jaglan A, Shaheen A, Yadav J, Lathwal P. Production of poly-β-hydroxybutyrate
(PHB) by bacteria isolated from rhizospheric soils. Int J Microbial Res Technol. 2015; 2(3): 38
-48.
17. Doetsch RN. Determinative methods of light microscopy. In: erhardt P, Murray R E,
Costilow RN, Nester EW, Wood WA, rieg NR, Phillips B. Manual of methods for general
bacteriology. 2nd ed. Washington. American Society of Microbiology; 1981: 22-31.
18. arrity M, Brenner DJ, rieg NR, Staley JT. Bergey's manual of systematic
bacteriology. 3nd ed. New York. Springer; 2005.
19. Vashist H, Sharma D, upta A. A review on commonly used biochemical test for bacteria.
Inov J Life Sci. 2013; 1(1): 1-7.
20. Ranganadha Reddy A, Venkateswarulu TC, Sudhakar P, rupanidhi S, Vidya Prabhakar .
Optimization of process parameters for polyhydroxybutyrate production from isolated
Acinetobacter nosocomialis RR20 through submerged fermentation. Current Trends in
Biotechnol Pharmacy. 2018; 12 (2): 159-168.
21. Zohri AN, El-Dean AM , Abuo-Dobara MI, Ali MI, Bakr MN, Ramy A H. Production of
polyhydroxyalkanoate by local strain of Bacillus egateriu AUMC b 272 utilizing sugar
beet wastewater and molasses. Egyptian Sugar J. 2019; 13: 45-70.
22. Chandani N, Mazumder PB, Bhattacharjee A. Biosynthesis of biodegradable polymer by a
potent soil bacterium from a stress-prone environment. J Applied Biology Biotechnol. 2018; 6
(2): 54-60.
23. El- Hamshary OIM, adi HA, AI-Twaty NH. Molecular characterization and uv improvement
of some bioplastic-producing bacteria isolated from plants in Taif city, Saudi Arabia.
Pharmacophore. 2018; 9(2): 7-18.
24. Hawas JM, El-Banna T, Belal EBA, El-Aziz AA. Production of bioplastic from some selected
bacterial strains. Int J Curr Microbiol App Sci. 2016; 5(1): 10-22.
25. Tamer TM, Omer AM, Hassan MA, Hassan ME, Sabet MM, Eldin MSM. Development
of thermos-sensittive poly N- isopropyl acrylamide grafted chitosan derivatives. J Applied
Pharmaceutical Sci. 2015; 5(3): 1-6.
26. Hassan MA, Bakhiet E , Salah A, Hussien HR. Production and characterization of
polyhydroxybutyrate (PHB) produced by Bacillus sp. isolated from Egypt. J Applied
Pharmaceutical Sci. 2016; 6(4): 46-51.
27. Weisburg W , Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for
phylogenetic study. J Bacteriol. 1991;173: 697–703.
28. Ahmed T, Shahid M, Azeem F, Rasul I, Shah AA, Noman M, Hameed A, Manzoor N,
Muhammad S. Biodegradation of plastics: current scenario and future prospects for
environmental safety. Environ Sci Pollut Res. 2018; 25(8):7287-7298.
29. Mascarenhas J, Aruna . Screening of polyhydroxyalkonates (PHA) accumulating
bacteria from diverse habitats. J lobal Biosci. 2017; 6: 4835-4848.
30. eshmiri S, Pordel S, Raeesi AR, Nabipour I, Darabi H, Jamali S, Dobaradaran S, Heidari
H, Ostovar A, Ramavandi B, Tahmasebi R, Marzban M, hajeian AM, Sanati AM, Farrokhi
Sh. Environmental pollution caused by gas and petrochemical industries and its effects on the
health of residents of Assaluyeh region, iranian energy capital: a review study. Iran South Med
J. 2018; 21(2): 162-185. [In Persian]
31. Desouky SE, El-Shiekh HH, Elabd MA, Shehab AM. Screening, optimization and extraction
of polyhydroxyalkanoates (PHAs) from Bacillus thuringienesis. J of Advances in Biology and
Biotechonol. 2014; 1(1): 40-54.
32. Thirumala M , Reddy SV, Mahmood S . Production and characterization of PHB from two
novel strains of Bacillus spp. isolated from soil and activated sludge. J Ind Microbiol
Biotechnol. 2010; 37(3):271–278.
33. Rostamzad A, Rezaee H, Hoshmndfar R. Isolation of biodegradable plastic producing bacteria
from soil contaminated sewage in Ilam milk manufactory. J Ilam Univ Med Sci. 2017; 25(3):
125-138. [In Persian]
34. Iman H , Nadhim HH, Saad H H. Bioplastic (poly -3-hydroxybutyrate) production by local
seudo onas aeruginosa isolates utilizing waste cooking oil. World J Pharma Res. 2017; 6
(8): 289-302.
35. Alshehrei F. Production of polyhydroxybutyrate (PHB) by bacteria isolated from soil of Saudi
Arabia. J Pure Appl Microbiol. 2019; 13(2): 897-904.
36. Bharathi B, owdhaman D, Ponnusami V. Isolation and identification of polyhydroxybutyrate
(PHB) producing Bacillus cereus BB613-A novel isolate. Int J ChemTech Res. 2016; 9(11):
224-228.
37. Sneha P, Changam SS, Balakumuran MD. Optimization, purification and characterization of
polyhydroxybutyrate (PHB) produced by Bacillus cereus isolated from sewage. Int J Chem
Tech Res. 2017; 10(7): 884-904.
38. Belal EB, Farid MA. Production of poly-β-hydroxybutyric acid (PHB) by Bacillus cereus. Int J
Curr Microbial App Sci. 2016; 5(7): 442-460.
39. Irsath H, Santhosh S, Hemalatha V, Vikramathithan M, Dhanasekar R, Dhandapani R. Production and optimization of polyhydroxybutyrate using Bacillus subtilis BP1 isolated from sewage
sample. Int J Pure App BioSci. 2015; 3(1): 158-166.
40. Hamieh A, Olama Z, Holail H. Microbial production of polyhydroxybutyrate, a biodegradable
plastic using agro-industrial waste products. lo Adv Res J Microbiol . 2013; 2(3): 54-64.
_||_
362- 367.
2. Shivalkar Y , Prabha R. Polyhydroxybutyrate as bio-degradable plastic – a review. J Environ
Sci Tox Food Technol. 2017, 11 (5): 10-12.
3. Sharma M, Dhingra H . Poly-β-hydroxybutyrate: a biodegradable polymer, biosynthesis and
biodegradation. BMJR. 2016; 14(3): 1-11.
4. Vijaya C, Reddy RM. Impact of soil composting using municipal solid waste on
biodegradation of plastics. Indian J Biotechnol. 2008; 7: 23 –239.
5. Reddy RL, Reddy VS, upta A. Study of bio-plastics as green and sustainable alternative to
plastics. Int J of Emerging Technol and Advanced Eng. 2013; 3(5): 82-89.
6. etachew A, Woldesenbet F. Production of biodegradable plastic by polyhydroxybutyrate
(PHB) accumulating bacteria using low cost agricultural waste material. BMC Res
Notes. 2016; 9(509): 1-9.
7. Al- addo HB, Sudesh , Samian MR. Screening of bacteria for PHA production using
waste glycerol as carbon source and the ability of new strain to produce P(3HB-co-3HV)
copolymer. Mal J Microbiol. 2016; 12(3): 245-253.
8. Panchal Sh. Recent developments on biodegradable polymers and their future trends. Int Res J
Sci Eng. 2016; 4 (1): 17-26.
9. Sharma M, Dhingra H . Poly-β-hydroxybutyrate: a biodegradable polyester, biosynthesis and
biodegradation. British Microbiology Res J. 2016; 14(3): 1-11.
10. Chaitanya , Mahmood S , ausar R, Sunilkumar N. Biotechnological production of
polyhydroxyalkanoates by various isolates: a review. Int J Pharma Sci Invent. 2014; 3(9):
111.
11. UMA Maheswari M, Dhandayuthapani . Fermentation optimization for production of
polyhydroxybutyrate (PHB) by newly isolated Azotobacter vinelandii DP. Int J Pharm
BioSci. 2013; 4(3): 779-787.
12. Shah R. Optimization and production of polyhydroxybutyrate (PHB) by Bacillus subtilis
1S1 from soil. Int J Curr Microbiol App Sci. 2014; 3(5): 377-387.
13. Jangra MR, Nehra S. Isolation, screening and characterization of new strains with
optimization studies to augment bacterial PHB production. Bull Env Pharmacol Life Sci.
2017; 6(8): 34-44.
14. Nehra , Chhabra N, Sidhu P , Lathwal P, Rana JS. Molecular identification and
characterization of poly-β-hydroxybutyrate (PHB) producing bacteria isolated from
contaminated soils. Asian J Microbiol Biotech Env Sci. 2015; 17(4): 281-290.
15. Bhagowati P, Pradhan Sh, Hirak R, Surajit D. Production, optimization and characterization
of polyhydroxybutyrate, a biodegradable plastic by Bacillus spp. Biosci Biotech
Biochem. 2015; 79(9): 1454- 1463.
16. Nehra , Jaglan A, Shaheen A, Yadav J, Lathwal P. Production of poly-β-hydroxybutyrate
(PHB) by bacteria isolated from rhizospheric soils. Int J Microbial Res Technol. 2015; 2(3): 38
-48.
17. Doetsch RN. Determinative methods of light microscopy. In: erhardt P, Murray R E,
Costilow RN, Nester EW, Wood WA, rieg NR, Phillips B. Manual of methods for general
bacteriology. 2nd ed. Washington. American Society of Microbiology; 1981: 22-31.
18. arrity M, Brenner DJ, rieg NR, Staley JT. Bergey's manual of systematic
bacteriology. 3nd ed. New York. Springer; 2005.
19. Vashist H, Sharma D, upta A. A review on commonly used biochemical test for bacteria.
Inov J Life Sci. 2013; 1(1): 1-7.
20. Ranganadha Reddy A, Venkateswarulu TC, Sudhakar P, rupanidhi S, Vidya Prabhakar .
Optimization of process parameters for polyhydroxybutyrate production from isolated
Acinetobacter nosocomialis RR20 through submerged fermentation. Current Trends in
Biotechnol Pharmacy. 2018; 12 (2): 159-168.
21. Zohri AN, El-Dean AM , Abuo-Dobara MI, Ali MI, Bakr MN, Ramy A H. Production of
polyhydroxyalkanoate by local strain of Bacillus egateriu AUMC b 272 utilizing sugar
beet wastewater and molasses. Egyptian Sugar J. 2019; 13: 45-70.
22. Chandani N, Mazumder PB, Bhattacharjee A. Biosynthesis of biodegradable polymer by a
potent soil bacterium from a stress-prone environment. J Applied Biology Biotechnol. 2018; 6
(2): 54-60.
23. El- Hamshary OIM, adi HA, AI-Twaty NH. Molecular characterization and uv improvement
of some bioplastic-producing bacteria isolated from plants in Taif city, Saudi Arabia.
Pharmacophore. 2018; 9(2): 7-18.
24. Hawas JM, El-Banna T, Belal EBA, El-Aziz AA. Production of bioplastic from some selected
bacterial strains. Int J Curr Microbiol App Sci. 2016; 5(1): 10-22.
25. Tamer TM, Omer AM, Hassan MA, Hassan ME, Sabet MM, Eldin MSM. Development
of thermos-sensittive poly N- isopropyl acrylamide grafted chitosan derivatives. J Applied
Pharmaceutical Sci. 2015; 5(3): 1-6.
26. Hassan MA, Bakhiet E , Salah A, Hussien HR. Production and characterization of
polyhydroxybutyrate (PHB) produced by Bacillus sp. isolated from Egypt. J Applied
Pharmaceutical Sci. 2016; 6(4): 46-51.
27. Weisburg W , Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for
phylogenetic study. J Bacteriol. 1991;173: 697–703.
28. Ahmed T, Shahid M, Azeem F, Rasul I, Shah AA, Noman M, Hameed A, Manzoor N,
Muhammad S. Biodegradation of plastics: current scenario and future prospects for
environmental safety. Environ Sci Pollut Res. 2018; 25(8):7287-7298.
29. Mascarenhas J, Aruna . Screening of polyhydroxyalkonates (PHA) accumulating
bacteria from diverse habitats. J lobal Biosci. 2017; 6: 4835-4848.
30. eshmiri S, Pordel S, Raeesi AR, Nabipour I, Darabi H, Jamali S, Dobaradaran S, Heidari
H, Ostovar A, Ramavandi B, Tahmasebi R, Marzban M, hajeian AM, Sanati AM, Farrokhi
Sh. Environmental pollution caused by gas and petrochemical industries and its effects on the
health of residents of Assaluyeh region, iranian energy capital: a review study. Iran South Med
J. 2018; 21(2): 162-185. [In Persian]
31. Desouky SE, El-Shiekh HH, Elabd MA, Shehab AM. Screening, optimization and extraction
of polyhydroxyalkanoates (PHAs) from Bacillus thuringienesis. J of Advances in Biology and
Biotechonol. 2014; 1(1): 40-54.
32. Thirumala M , Reddy SV, Mahmood S . Production and characterization of PHB from two
novel strains of Bacillus spp. isolated from soil and activated sludge. J Ind Microbiol
Biotechnol. 2010; 37(3):271–278.
33. Rostamzad A, Rezaee H, Hoshmndfar R. Isolation of biodegradable plastic producing bacteria
from soil contaminated sewage in Ilam milk manufactory. J Ilam Univ Med Sci. 2017; 25(3):
125-138. [In Persian]
34. Iman H , Nadhim HH, Saad H H. Bioplastic (poly -3-hydroxybutyrate) production by local
seudo onas aeruginosa isolates utilizing waste cooking oil. World J Pharma Res. 2017; 6
(8): 289-302.
35. Alshehrei F. Production of polyhydroxybutyrate (PHB) by bacteria isolated from soil of Saudi
Arabia. J Pure Appl Microbiol. 2019; 13(2): 897-904.
36. Bharathi B, owdhaman D, Ponnusami V. Isolation and identification of polyhydroxybutyrate
(PHB) producing Bacillus cereus BB613-A novel isolate. Int J ChemTech Res. 2016; 9(11):
224-228.
37. Sneha P, Changam SS, Balakumuran MD. Optimization, purification and characterization of
polyhydroxybutyrate (PHB) produced by Bacillus cereus isolated from sewage. Int J Chem
Tech Res. 2017; 10(7): 884-904.
38. Belal EB, Farid MA. Production of poly-β-hydroxybutyric acid (PHB) by Bacillus cereus. Int J
Curr Microbial App Sci. 2016; 5(7): 442-460.
39. Irsath H, Santhosh S, Hemalatha V, Vikramathithan M, Dhanasekar R, Dhandapani R. Production and optimization of polyhydroxybutyrate using Bacillus subtilis BP1 isolated from sewage
sample. Int J Pure App BioSci. 2015; 3(1): 158-166.
40. Hamieh A, Olama Z, Holail H. Microbial production of polyhydroxybutyrate, a biodegradable
plastic using agro-industrial waste products. lo Adv Res J Microbiol . 2013; 2(3): 54-64.