Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome
Abstract Background Lignocellulose biomass is the most abundant and renewable material in nature. The objectives of this study were to characterize two endoglucanases TrepCel3 and TrepCel4, and determine the effect of the combination of them (1.2 mg TrepCel3, 0.8 mg TrepCel4) on in vitro rumen ferme...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-02-01
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| Series: | Journal of Animal Science and Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40104-022-00814-z |
| _version_ | 1811165770359504896 |
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| author | Zhenxiang Meng Chengjian Yang Jing Leng Weiyun Zhu Yanfen Cheng |
| author_facet | Zhenxiang Meng Chengjian Yang Jing Leng Weiyun Zhu Yanfen Cheng |
| author_sort | Zhenxiang Meng |
| collection | DOAJ |
| description | Abstract Background Lignocellulose biomass is the most abundant and renewable material in nature. The objectives of this study were to characterize two endoglucanases TrepCel3 and TrepCel4, and determine the effect of the combination of them (1.2 mg TrepCel3, 0.8 mg TrepCel4) on in vitro rumen fermentation characteristics. In this study, three nature lignocellulosic substrates (rice straw, RS; wheat straw, WS; leymus chinensis, LC) were evaluated for their in vitro digestibility, gas, NH3-N and volatile fatty acid (VFA) production, and microbial protein (MCP) synthesis by adding enzymatic combination. Methods Two endoglucanases’ genes were successfully expressed in Escherichia coli (E. coli) BL21 (DE3), and enzymatic characteristics were further characterized. The combination of TrepCel3 and TrepCel4 was incubated with lignocellulosic substrates to evaluate its hydrolysis ability. Results The maximum enzymatic activity of TrepCel3 was determined at pH 5.0 and 40 °C, while TrepCel4 was at pH 6.0 and 50 °C. They were stable over the temperature range of 30 to 60 °C, and active within the pH range of 4.0 to 9.0. The TrepCel3 and TrepCel4 had the highest activity in lichenan 436.9 ± 8.30 and 377.6 ± 6.80 U/mg, respectively. The combination of TrepCel3 and TrepCel4 exhibited the highest efficiency at the ratio of 60:40. Compared to maximum hydrolysis of TrepCel3 or TrepCel4 separately, this combination was shown to have a superior ability to maximize the saccharification yield from lignocellulosic substrates up to 188.4% for RS, 236.7% for wheat straw WS, 222.4% for LC and 131.1% for sugar beet pulp (SBP). Supplemental this combination enhanced the dry matter digestion (DMD), gas, NH3-N and VFA production, and MCP synthesis during in vitro rumen fermentation. Conclusions The TrepCel3 and TrepCel4 exhibited the synergistic relationship (60:40) and significantly increased the saccharification yield of lignocellulosic substrates. The combination of them stimulated in vitro rumen fermentation of lignocellulosic substrates. This combination has the potential to be a feed additive to improve agricultural residues utilization in ruminants. If possible, in the future, experiments in vivo should be carried out to fully evaluate its effect. |
| first_indexed | 2024-04-10T15:41:57Z |
| format | Article |
| id | doaj.art-511d43aa282d44449b916a66f182376b |
| institution | Directory Open Access Journal |
| issn | 2049-1891 |
| language | English |
| last_indexed | 2024-04-10T15:41:57Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Animal Science and Biotechnology |
| spelling | doaj.art-511d43aa282d44449b916a66f182376b2023-02-12T12:18:53ZengBMCJournal of Animal Science and Biotechnology2049-18912023-02-0114111710.1186/s40104-022-00814-zProduction, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenomeZhenxiang Meng0Chengjian Yang1Jing Leng2Weiyun Zhu3Yanfen Cheng4Laboratory of Gastrointestinal Microbiology, National Center for International Research On Animal Gut Nutrition, Nanjing Agricultural UniversityBuffalo Research Institute, Chinese Academy of AgriculturalKey Laboratory of Animal Nutrition and Feed Science of Yunnan Province, Yunnan Agricultural UniversityLaboratory of Gastrointestinal Microbiology, National Center for International Research On Animal Gut Nutrition, Nanjing Agricultural UniversityLaboratory of Gastrointestinal Microbiology, National Center for International Research On Animal Gut Nutrition, Nanjing Agricultural UniversityAbstract Background Lignocellulose biomass is the most abundant and renewable material in nature. The objectives of this study were to characterize two endoglucanases TrepCel3 and TrepCel4, and determine the effect of the combination of them (1.2 mg TrepCel3, 0.8 mg TrepCel4) on in vitro rumen fermentation characteristics. In this study, three nature lignocellulosic substrates (rice straw, RS; wheat straw, WS; leymus chinensis, LC) were evaluated for their in vitro digestibility, gas, NH3-N and volatile fatty acid (VFA) production, and microbial protein (MCP) synthesis by adding enzymatic combination. Methods Two endoglucanases’ genes were successfully expressed in Escherichia coli (E. coli) BL21 (DE3), and enzymatic characteristics were further characterized. The combination of TrepCel3 and TrepCel4 was incubated with lignocellulosic substrates to evaluate its hydrolysis ability. Results The maximum enzymatic activity of TrepCel3 was determined at pH 5.0 and 40 °C, while TrepCel4 was at pH 6.0 and 50 °C. They were stable over the temperature range of 30 to 60 °C, and active within the pH range of 4.0 to 9.0. The TrepCel3 and TrepCel4 had the highest activity in lichenan 436.9 ± 8.30 and 377.6 ± 6.80 U/mg, respectively. The combination of TrepCel3 and TrepCel4 exhibited the highest efficiency at the ratio of 60:40. Compared to maximum hydrolysis of TrepCel3 or TrepCel4 separately, this combination was shown to have a superior ability to maximize the saccharification yield from lignocellulosic substrates up to 188.4% for RS, 236.7% for wheat straw WS, 222.4% for LC and 131.1% for sugar beet pulp (SBP). Supplemental this combination enhanced the dry matter digestion (DMD), gas, NH3-N and VFA production, and MCP synthesis during in vitro rumen fermentation. Conclusions The TrepCel3 and TrepCel4 exhibited the synergistic relationship (60:40) and significantly increased the saccharification yield of lignocellulosic substrates. The combination of them stimulated in vitro rumen fermentation of lignocellulosic substrates. This combination has the potential to be a feed additive to improve agricultural residues utilization in ruminants. If possible, in the future, experiments in vivo should be carried out to fully evaluate its effect.https://doi.org/10.1186/s40104-022-00814-zEndoglucanaseIn vitroLignocellulosic substratesRumen fermentation |
| spellingShingle | Zhenxiang Meng Chengjian Yang Jing Leng Weiyun Zhu Yanfen Cheng Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome Journal of Animal Science and Biotechnology Endoglucanase In vitro Lignocellulosic substrates Rumen fermentation |
| title | Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| title_full | Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| title_fullStr | Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| title_full_unstemmed | Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| title_short | Production, purification, characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| title_sort | production purification characterization and application of two novel endoglucanases from buffalo rumen metagenome |
| topic | Endoglucanase In vitro Lignocellulosic substrates Rumen fermentation |
| url | https://doi.org/10.1186/s40104-022-00814-z |
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