q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung
This study aims to create a quantitative polymerase chain reaction (q-PCR) methodology for monitoring the hydrogen-producing mixed cultures enriched from elephant dung using alpha-cellulose as a carbon source through five generations of repetitive sub-culture. The enriched thermophilic mixed culture...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/10/506 |
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| author | Khamanitjaree Saripan Chonticha Mamimin Tsuyoshi Imai Sureewan Sittijunda Alissara Reungsang |
| author_facet | Khamanitjaree Saripan Chonticha Mamimin Tsuyoshi Imai Sureewan Sittijunda Alissara Reungsang |
| author_sort | Khamanitjaree Saripan |
| collection | DOAJ |
| description | This study aims to create a quantitative polymerase chain reaction (q-PCR) methodology for monitoring the hydrogen-producing mixed cultures enriched from elephant dung using alpha-cellulose as a carbon source through five generations of repetitive sub-culture. The enriched thermophilic mixed cultures from the fifth cultivation cycle gave the highest hydrogen yield of 170.3 mL H<sub>2</sub>/g cellulose and were used to generate hydrogen from sawdust. <i>Clostridium</i> sp. and <i>Thermoanaerobacterium</i> sp. were the dominant bacteria in thermophilic mixed cultures with high hydrogen yield, according to polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE). q-PCR primers Chis150F and ClostIR, TherF and TherR, and BacdF and BacdR were developed to amplify the 16S rRNA genes of <i>Clostridium</i> sp., <i>Thermoanaerobacterium</i> sp., and <i>Bacillus</i> sp., respectively, for the quantification of hydrogen-producing bacteria in biohydrogen fermentation. Similar q-PCR analysis of <i>Clostridium</i> sp., <i>Thermoanaerobacterium</i> sp., and <i>Bacillus</i> sp. 16S rRNA gene amplification during hydrogen production from cellulose and sawdust revealed increasing gene copy number with time. The molecular approaches developed in this study can be used to monitor microbial communities in hydrogen fermentation processes efficiently. |
| first_indexed | 2024-03-09T12:19:46Z |
| format | Article |
| id | doaj.art-57804af3ddd14d4d9799761eda1a1bd5 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-09T12:19:46Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-57804af3ddd14d4d9799761eda1a1bd52023-11-30T22:42:01ZengMDPI AGFermentation2311-56372022-10-0181050610.3390/fermentation8100506q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant DungKhamanitjaree Saripan0Chonticha Mamimin1Tsuyoshi Imai2Sureewan Sittijunda3Alissara Reungsang4Environmental Science Program, Faculty of Science and Technology, Thepsatri Rajabhat University, A. Muang, Lopburi 40002, ThailandDepartment of Biotechnology, Faculty of Technology, Khon Kaen University, A. Muang, Khon Kaen 40002, ThailandGraduate School of Sciences and Technology for Innovation, Yamaguchi University, Yamaguchi 755-8611, JapanFaculty of Environment and Resource Studies, Mahidol University, Nakhon Pathom 73170, ThailandDepartment of Biotechnology, Faculty of Technology, Khon Kaen University, A. Muang, Khon Kaen 40002, ThailandThis study aims to create a quantitative polymerase chain reaction (q-PCR) methodology for monitoring the hydrogen-producing mixed cultures enriched from elephant dung using alpha-cellulose as a carbon source through five generations of repetitive sub-culture. The enriched thermophilic mixed cultures from the fifth cultivation cycle gave the highest hydrogen yield of 170.3 mL H<sub>2</sub>/g cellulose and were used to generate hydrogen from sawdust. <i>Clostridium</i> sp. and <i>Thermoanaerobacterium</i> sp. were the dominant bacteria in thermophilic mixed cultures with high hydrogen yield, according to polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE). q-PCR primers Chis150F and ClostIR, TherF and TherR, and BacdF and BacdR were developed to amplify the 16S rRNA genes of <i>Clostridium</i> sp., <i>Thermoanaerobacterium</i> sp., and <i>Bacillus</i> sp., respectively, for the quantification of hydrogen-producing bacteria in biohydrogen fermentation. Similar q-PCR analysis of <i>Clostridium</i> sp., <i>Thermoanaerobacterium</i> sp., and <i>Bacillus</i> sp. 16S rRNA gene amplification during hydrogen production from cellulose and sawdust revealed increasing gene copy number with time. The molecular approaches developed in this study can be used to monitor microbial communities in hydrogen fermentation processes efficiently.https://www.mdpi.com/2311-5637/8/10/506PCR-DGGEmicrobial communitythermophilic hydrogen productionsawdustlignocellulosic biomass |
| spellingShingle | Khamanitjaree Saripan Chonticha Mamimin Tsuyoshi Imai Sureewan Sittijunda Alissara Reungsang q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung Fermentation PCR-DGGE microbial community thermophilic hydrogen production sawdust lignocellulosic biomass |
| title | q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung |
| title_full | q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung |
| title_fullStr | q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung |
| title_full_unstemmed | q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung |
| title_short | q-PCR Methodology for Monitoring the Thermophilic Hydrogen Producers Enriched from Elephant Dung |
| title_sort | q pcr methodology for monitoring the thermophilic hydrogen producers enriched from elephant dung |
| topic | PCR-DGGE microbial community thermophilic hydrogen production sawdust lignocellulosic biomass |
| url | https://www.mdpi.com/2311-5637/8/10/506 |
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