تولید بوتانول زیستی از ضایعات نان با استفاده از کلستریدیوم‌های آمیلولیتکی جدا شده از دریاچه پریشان
محورهای موضوعی : میکروب شناسی
مریم میرزاده
1
(کارشناسی ارشد، گروه میکروبیولوژی، واحد جهرم، دانشگاه آزاد اسلامی، جهرم، ایران.)
عباسعلی رضائیان
2
(استادیار، گروه میکروبیولوژی، واحد جهرم، دانشگاه آزاد اسلامی، جهرم، ایران)
کلید واژه: ضایعات نان, کلستریدیم, آنزیم آمیلاز, بوتانول زیستی, دریاچه پریشان,
چکیده مقاله :
زمینه و هدف:بوتانول زیستی به دلیل تجدیدپذیر بودن، به عنوان یک جایگزین برای سوخت‌های نفتی مورد توجه قرار گرفته است. هدف این پژوهش استفاده از باکتری‌های محیطی و ضایعات نان جهت تولید بوتانول زیستی می‌باشد. روش‌شناسی:در این پژوهش، به طور تصادفی از عمق 50 تا ۱۰۰ سانتی متری رسوبات دریاچه پریشان نمونه تهیه شد. پس از کشت در شرایط بی‌هوازی و تشخیص فنوتیپی جنس کلستریدیوم، از پرایمر 16S rRNA جهت شناسایی ژنوتیپی استفاده گردید. جدایه‌ها براساس فعالیت آمیلازی در محیط کشت نشاسته-آگار غربالگری شدند. برای بررسی تأثیر فاکتورهای محیطی (pH، دما و ماده اولیه) بر فعالیت آمیلازی جدایه‌های منتخب، از یک محیط کشت تغییر شکل یافته حاوی نشاسته استفاده گردید. سپس در یک محیط کشت بهینه، صرفاً شامل آب و نان خشک، بوتانول زیستی تولید و بوسیله تقطیر جزء به جزء جداسازی شد. صحّت وجود آن با تست اسیدکرومیک و دستگاه کروماتوگرافی گازی تأیید گردید. یافته‌ها:از مجموع 530 باکتری جدا شده، 3 جدایه کلستریدیوم بیشترین فعالیت آمیلازی و تولید بوتانول زیستی را داشتند که پس از تعیین توالی 16S rRNA، با نام‌های کلستریدیوم بیجرینکی (KM999944)، کلستریدیوم دیولیس (KM999945) و کلستریدیوم روزئوم (KM999946) در NCBI ثبت گردیدند. بالاترین محصول با غلظت g/l344/2 در محیط کشت حاوی g/l25 ضایعات نان، 7pH و دمای 35 درجه سلسیوس مربوط به کلستریدیوم روزئوم بوده است. نتیجه‌گیری:نتایج نشان داد که در شرایط بهینه، کلستریدیوم‌های محیطی، پتانسیل خوبی برای تولید بوتانول زیستی از مواد اولیه ارزان قیمتی همچون ضایعات نان دارند.
Background and Aim: Bio-butanol has been regarded as an alternative to petroleum fuels due to its renewability. The aim of this study is to produce bio-butanol using bread waste and environmental bacteria. Materials and Methods: In this study, samples were randomly taken from the depth of 50 to 100 cm of Parishan Lake sediments. After culturing under anaerobic conditions and phenotypic diagnosis of the Clostridium genus, 16S rRNA primer was used for genotypic identification. Isolates were screened based on amylase activity in starch-agar medium. To evaluate the effect of environmental factors (pH, temperature and raw material) on amylase activity of the selected isolates, a modified culture medium containing starch was used. Then, in an optimal culture medium, merely consisting of water and dry bread, bio-butanol was produced and separated by fractional distillation. The accuracy of its presence was confirmed by chromic acid test and gas chromatography. Results: Out of a total of 530 isolated bacteria, three clostridium isolates had the highest amylase activity and bio-butanol production, which after 16S rRNA sequencing, were registered as Clostridium Beijerinckii (KM999944), Clostridium diolis (KM999945) and Clostridium roseum (KM999946) in NCBI. The highest yield with a concentration of 2.344g/l in the culture medium containing 25g/l bread waste, pH7 and 35°C has been related to Clostridium roseum. Conclusion: The results showed that under optimal conditions, environmental clostridia have a good potential for the production of bio-butanol from cheap raw materials like bread waste.
Gaurav N, Sivasankari S, Kiran GS, Ninawe A, Selvin J. Utilization of bioresources for sustainable biofuels: A Review. Renew Sust Energ Rev. 2017; 73: 205-14.
Seo PW, Ahmed I, Jhung SH. Adsorptive removal of nitrogen-containing compounds from a model fuel using a metal-organic framework having a free carboxylic acid group. Chem Eng J. 2016; 299: 236-43.
Rakopoulos DC, Rakopoulos CD, Kyritsis DC. Butanol or DEE blends with either straight vegetable oil or biodiesel excluding fossil fuel: Comparative effects on diesel engine combustion attributes, cyclic variability and regulated emissions trade-off. Energy. 2016; 115: 314-25.
Liu C-M, Wu S-Y. From biomass waste to biofuels and biomaterial building blocks. Renew Energy. 2016; 96: 1056-62.
Rodionova MV, Poudyal RS, Tiwari I, Voloshin RA, Zharmukhamedov SK, Nam HG & et al. Biofuel production: challenges and opportunities. Int J Hydrogen Energy. 2017; 42(12): 8450-8461.
Whalen J, Xu CC, Shen F, Kumar A, Eklund M, Yan J. Sustainable biofuel production from forestry, agricultural and waste biomass feedstocks. Appl Energy. 2017; 198:
281-283.
Zabed H, Sahu JN, Suely A, Boyce AN, Faruq G. Bioethanol production from renewable sources: Current perspectives and technological progress. Renew Sust Energ Rev. 2017; 71: 475-501.
Cardona CA, Sánchez ÓJ. Fuel ethanol production: process design trends and integration opportunities. Bioresour Technol.2007; 98(12): 2415-2457.
Bharathiraja B, Jayamuthunagai J, Sudharsanaa T, Bharghavi A, Praveenkumar R, Chakravarthy M & et al. Biobutanol-An impending biofuel for future: A review on upstream and downstream processing tecniques. Renew Sust Energ Rev. 2017; 68:
788-807.
Rathour RK, Ahuja V, Bhatia RK, Bhatt AK. Biobutanol: New era of biofuels. Int J Energy Res. 2018; 42(15): 4532-45.
Feng H, Liu D, Yang X, An M, Zhang W, Zhang X. Availability analysis of using
iso-octane/n-butanol blends in spark-ignition engines. Renew Energy. 2016; 96: 281-94.
Da Silva Trindade WR, dos Santos RG. Review on the characteristics of butanol, its production and use as fuel in internal combustion engines. Renew Sust Energ Rev. 2017; 69: 642-51.
Brito M, Martins F. Life cycle assessment of butanol production. Fuel. 2017; 208: 476-82.
Jiménez-Bonilla P, Wang Y. In situ biobutanol recovery from clostridial fermentations: a critical review. Crit Rev Biotechnol. 2018; 38(3): 469-82.
Millat T, Winzer K. Mathematical modelling of clostridial acetone-butanol-ethanol fermentation. Appl Microbiol Biotechnol. 2017; 101(6): 2251-71.
Zhang Y-HP, Sun J, Ma Y. Biomanufacturing: history and perspective. J Ind Microbiol Biotechnol. 2017; 44(4-5): 773-84.
Procentese A, Raganati F, Olivieri G, Russo ME, de la Feld M, Marzocchella A. Renewable feedstocks for biobutanol production by fermentation. N Biotechnol. 2017;
39: 135-40.
Maiti S, Sarma SJ, Brar SK, Le Bihan Y, Drogui P, Buelna G & et al. Agro-industrial wastes as feedstock for sustainable bio-production of butanol by Clostridium beijerinckii. Food Bioprod Process. 2016; 98: 217-26.
Leung CCJ, Cheung ASY, Zhang AY-Z, Lam KF, Lin CSK. Utilisation of waste bread for fermentative succinic acid production. Biochem Eng J. 2012; 65: 10-5.
Vos PD, Garrity GM, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, Whitman WB. Bergey's Manual of Systematic Bacteriology. Springer. 2009: 738-828
Fuwa H. A new method for microdetermination CF amylase activity by the use of amylose as the substrate. J Biochem. 1954; 41(5): 583-603.
Abdollahi M, Karbalaei-Heidari H. Isolation, identification, biochemical properties and potential application of an organic solvent-tolerant lipase from Pseudomonas sp. strain NEB-1. Iran J Sci Technol Trans A Sci. 2014; 38(3): 221-9.
Smith BW, Roe JH. A photometric method for the determination of α-amylase in blood and urine, with use of the starch-iodine color. J Biol Chem.1949; 179(1): 53-9.
Bohringer P, Jacob A. The determination of alcohol using chromic acid. Zeitschr Flussiges Abst. 1964; 31: 233-6.
Qin Z, Duns GJ, Pan T, Xin F. Consolidated processing of biobutanol production from food wastes by solventogenic Clostridium sp. strain HN4. Bioresour Technol. 2018; 264: 148-53.
Luo W, Zhao Z, Pan H, Zhao L, Xu C, Yu X. Feasibility of butanol production from wheat starch wastewater by Clostridium acetobutylicum. Energy. 2018; 154: 240-8.
Husin H, Ibrahim MF, Kamal Bahrin E, Abd‐Aziz S. Simultaneous saccharification and fermentation of sago hampas into biobutanol by Clostridium acetobutylicumATCC 824. Energy Sci Eng. 2018;7(1): 66-75.
Johnravindar D, Elangovan N, Gopal NO, Muthaiyan A, Fei Q. Biobutanol production from cassava waste residue using Clostridium sp. AS3 in batch culture fermentation. Biofuels. 2019:1-8. DOI: https://doi.org/10.1080/17597269.2019.1608671.
Lin Z, Liu H, Wu J, Patakova P, Branska B, Zhang J. Effective continuous acetone-butanol-ethanol production with full utilization of cassava by immobilized symbiotic TSH06. Biotechnol Biofuels. 2019; 12(219): 1-11.
Yang M, Kuittinen S, Zhang J, Vepsäläinen J, Keinänen M, Pappinen A. Co-fermentation of hemicellulose and starch from barley straw and grain for efficient pentoses utilization in acetone-butanol-ethanol production. Bioresour Technol. 2015; 179: 128-35.
Badr HR, Toledo R, Hamdy MK. Continuous acetone-ethanol-butanol fermentation
by immobilized cells of Clostridium acetobutylicum. Biomass Bioenergy. 2001; 20(2):
119-32.
Li S, Guo Y, Lu F, Huang J, Pang Z. High-level butanol production from cassava starch by a newly isolated Clostridium acetobutylicum. Appl Biochem Biotechnol. 2015; 177(4): 831-41.
Al-Shorgani NKN, Kalil MS, Yusoff WMW. Fermentation of sago starch to biobutanol in a batch culture using Clostridium saccharoperbutylacetonicumN1-4 (ATCC 13564). Ann Microbiol. 2012; 62(3): 1059-70.
Ding J, Xu M, Xie F, Chen C, Shi Z. Efficient butanol production using corn-starch and waste Pichia pastoris semi-solid mixture as the substrate. Biochem Eng J. 2019; 143:
41-7.
Gaurav N, Sivasankari S, Kiran GS, Ninawe A, Selvin J. Utilization of bioresources for sustainable biofuels: A Review. Renew Sust Energ Rev. 2017; 73: 205-14.
Seo PW, Ahmed I, Jhung SH. Adsorptive removal of nitrogen-containing compounds from a model fuel using a metal-organic framework having a free carboxylic acid group. Chem Eng J. 2016; 299: 236-43.
Rakopoulos DC, Rakopoulos CD, Kyritsis DC. Butanol or DEE blends with either straight vegetable oil or biodiesel excluding fossil fuel: Comparative effects on diesel engine combustion attributes, cyclic variability and regulated emissions trade-off. Energy. 2016; 115: 314-25.
Liu C-M, Wu S-Y. From biomass waste to biofuels and biomaterial building blocks. Renew Energy. 2016; 96: 1056-62.
Rodionova MV, Poudyal RS, Tiwari I, Voloshin RA, Zharmukhamedov SK, Nam HG & et al. Biofuel production: challenges and opportunities. Int J Hydrogen Energy. 2017; 42(12): 8450-8461.
Whalen J, Xu CC, Shen F, Kumar A, Eklund M, Yan J. Sustainable biofuel production from forestry, agricultural and waste biomass feedstocks. Appl Energy. 2017; 198:
281-283.
Zabed H, Sahu JN, Suely A, Boyce AN, Faruq G. Bioethanol production from renewable sources: Current perspectives and technological progress. Renew Sust Energ Rev. 2017; 71: 475-501.
Cardona CA, Sánchez ÓJ. Fuel ethanol production: process design trends and integration opportunities. Bioresour Technol.2007; 98(12): 2415-2457.
Bharathiraja B, Jayamuthunagai J, Sudharsanaa T, Bharghavi A, Praveenkumar R, Chakravarthy M & et al. Biobutanol-An impending biofuel for future: A review on upstream and downstream processing tecniques. Renew Sust Energ Rev. 2017; 68:
788-807.
Rathour RK, Ahuja V, Bhatia RK, Bhatt AK. Biobutanol: New era of biofuels. Int J Energy Res. 2018; 42(15): 4532-45.
Feng H, Liu D, Yang X, An M, Zhang W, Zhang X. Availability analysis of using
iso-octane/n-butanol blends in spark-ignition engines. Renew Energy. 2016; 96: 281-94.
Da Silva Trindade WR, dos Santos RG. Review on the characteristics of butanol, its production and use as fuel in internal combustion engines. Renew Sust Energ Rev. 2017; 69: 642-51.
Brito M, Martins F. Life cycle assessment of butanol production. Fuel. 2017; 208: 476-82.
Jiménez-Bonilla P, Wang Y. In situ biobutanol recovery from clostridial fermentations: a critical review. Crit Rev Biotechnol. 2018; 38(3): 469-82.
Millat T, Winzer K. Mathematical modelling of clostridial acetone-butanol-ethanol fermentation. Appl Microbiol Biotechnol. 2017; 101(6): 2251-71.
Zhang Y-HP, Sun J, Ma Y. Biomanufacturing: history and perspective. J Ind Microbiol Biotechnol. 2017; 44(4-5): 773-84.
Procentese A, Raganati F, Olivieri G, Russo ME, de la Feld M, Marzocchella A. Renewable feedstocks for biobutanol production by fermentation. N Biotechnol. 2017;
39: 135-40.
Maiti S, Sarma SJ, Brar SK, Le Bihan Y, Drogui P, Buelna G & et al. Agro-industrial wastes as feedstock for sustainable bio-production of butanol by Clostridium beijerinckii. Food Bioprod Process. 2016; 98: 217-26.
Leung CCJ, Cheung ASY, Zhang AY-Z, Lam KF, Lin CSK. Utilisation of waste bread for fermentative succinic acid production. Biochem Eng J. 2012; 65: 10-5.
Vos PD, Garrity GM, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, Whitman WB. Bergey's Manual of Systematic Bacteriology. Springer. 2009: 738-828
Fuwa H. A new method for microdetermination CF amylase activity by the use of amylose as the substrate. J Biochem. 1954; 41(5): 583-603.
Abdollahi M, Karbalaei-Heidari H. Isolation, identification, biochemical properties and potential application of an organic solvent-tolerant lipase from Pseudomonas sp. strain NEB-1. Iran J Sci Technol Trans A Sci. 2014; 38(3): 221-9.
Smith BW, Roe JH. A photometric method for the determination of α-amylase in blood and urine, with use of the starch-iodine color. J Biol Chem.1949; 179(1): 53-9.
Bohringer P, Jacob A. The determination of alcohol using chromic acid. Zeitschr Flussiges Abst. 1964; 31: 233-6.
Qin Z, Duns GJ, Pan T, Xin F. Consolidated processing of biobutanol production from food wastes by solventogenic Clostridium sp. strain HN4. Bioresour Technol. 2018; 264: 148-53.
Luo W, Zhao Z, Pan H, Zhao L, Xu C, Yu X. Feasibility of butanol production from wheat starch wastewater by Clostridium acetobutylicum. Energy. 2018; 154: 240-8.
Husin H, Ibrahim MF, Kamal Bahrin E, Abd‐Aziz S. Simultaneous saccharification and fermentation of sago hampas into biobutanol by Clostridium acetobutylicumATCC 824. Energy Sci Eng. 2018;7(1): 66-75.
Johnravindar D, Elangovan N, Gopal NO, Muthaiyan A, Fei Q. Biobutanol production from cassava waste residue using Clostridium sp. AS3 in batch culture fermentation. Biofuels. 2019:1-8. DOI: https://doi.org/10.1080/17597269.2019.1608671.
Lin Z, Liu H, Wu J, Patakova P, Branska B, Zhang J. Effective continuous acetone-butanol-ethanol production with full utilization of cassava by immobilized symbiotic TSH06. Biotechnol Biofuels. 2019; 12(219): 1-11.
Yang M, Kuittinen S, Zhang J, Vepsäläinen J, Keinänen M, Pappinen A. Co-fermentation of hemicellulose and starch from barley straw and grain for efficient pentoses utilization in acetone-butanol-ethanol production. Bioresour Technol. 2015; 179: 128-35.
Badr HR, Toledo R, Hamdy MK. Continuous acetone-ethanol-butanol fermentation
by immobilized cells of Clostridium acetobutylicum. Biomass Bioenergy. 2001; 20(2):
119-32.
Li S, Guo Y, Lu F, Huang J, Pang Z. High-level butanol production from cassava starch by a newly isolated Clostridium acetobutylicum. Appl Biochem Biotechnol. 2015; 177(4): 831-41.
Al-Shorgani NKN, Kalil MS, Yusoff WMW. Fermentation of sago starch to biobutanol in a batch culture using Clostridium saccharoperbutylacetonicumN1-4 (ATCC 13564). Ann Microbiol. 2012; 62(3): 1059-70.
Ding J, Xu M, Xie F, Chen C, Shi Z. Efficient butanol production using corn-starch and waste Pichia pastoris semi-solid mixture as the substrate. Biochem Eng J. 2019; 143:
41-7.
