<i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications
Actinomycetes are an attractive source of lignocellulose-degrading enzymes. The search for actinomycetes producing extremozyme cellulase using cheap lignocellulosic waste remains a priority goal of enzyme research. In this context, the extremophilic actinomycete NBRM9 showed promising cellulolytic a...
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AIMS Press
2024-03-01
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| Series: | AIMS Microbiology |
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| Online Access: | https://www.aimspress.com/article/doi/10.3934/microbiol.2024010?viewType=HTML |
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| author | Mohamed H. El-Sayed Doaa A. Elsayed Abd El-Rahman F. Gomaa |
| author_facet | Mohamed H. El-Sayed Doaa A. Elsayed Abd El-Rahman F. Gomaa |
| author_sort | Mohamed H. El-Sayed |
| collection | DOAJ |
| description | Actinomycetes are an attractive source of lignocellulose-degrading enzymes. The search for actinomycetes producing extremozyme cellulase using cheap lignocellulosic waste remains a priority goal of enzyme research. In this context, the extremophilic actinomycete NBRM9 showed promising cellulolytic activity in solid and liquid assays. This actinomycete was identified as <i>Nocardiopsis synnemataformans</i> based on its phenotypic characteristics alongside phylogenetic analyses of 16S rRNA gene sequencing (OQ380604.1). Using bean straw as the best agro-waste, the production of cellulase from this strain was statistically optimized using a response surface methodology, with the maximum activity (13.20 U/mL) achieved at an incubation temperature of 40 °C, a pH of 9, an incubation time of 7 days, and a 2% substrate concentration. The partially purified cellulase (PPC) showed promising activity and stability over a wide range of temperatures (20–90 °C), pH values (3–11), and NaCl concentrations (1–19%), with optimal activity at 50 °C, pH 9.0, and 10% salinity. Under these conditions, the enzyme retained >95% of its activity, thus indicating its extremozyme nature. The kinetics of cellulase showed that it has a V<sub>max</sub> of 20.19 ± 1.88 U/mL and a Km of 0.25 ± 0.07 mM. The immobilized PPC had a relative activity of 69.58 ± 0.13%. In the in vitro microtiter assay, the PPC was found to have a concentration-dependent anti-biofilm activity (up to 85.15 ± 1.60%). Additionally, the fermentative conversion of the hydrolyzed bean straw by <i>Saccharomyces cerevisiae</i> (KM504287.1) amounted to 65.80 ± 0.52% of the theoretical ethanol yield. Overall, for the first time, the present work reports the production of extremozymatic (thermo, alkali-, and halo-stable) cellulase from <i>N. synnemataformans</i> NBRM9. Therefore, this strain is recommended for use as a biotool in many lignocellulosic-based applications operating under harsh conditions. |
| first_indexed | 2024-04-24T11:00:16Z |
| format | Article |
| id | doaj.art-f1961672e24945a6b921b333b30e1ce5 |
| institution | Directory Open Access Journal |
| issn | 2471-1888 |
| language | English |
| last_indexed | 2024-04-24T11:00:16Z |
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| spelling | doaj.art-f1961672e24945a6b921b333b30e1ce52024-04-12T01:35:07ZengAIMS PressAIMS Microbiology2471-18882024-03-0110118721910.3934/microbiol.2024010<i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applicationsMohamed H. El-Sayed0Doaa A. Elsayed 1Abd El-Rahman F. Gomaa21. Department of Biology, College of Science and Arts, Northern Border University, Arar, Saudi Arabia 2. Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Cairo 11884, Egypt1. Department of Biology, College of Science and Arts, Northern Border University, Arar, Saudi Arabia3. Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt 4. The Key Laboratory of Biotechnology for Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu, 221116, PR ChinaActinomycetes are an attractive source of lignocellulose-degrading enzymes. The search for actinomycetes producing extremozyme cellulase using cheap lignocellulosic waste remains a priority goal of enzyme research. In this context, the extremophilic actinomycete NBRM9 showed promising cellulolytic activity in solid and liquid assays. This actinomycete was identified as <i>Nocardiopsis synnemataformans</i> based on its phenotypic characteristics alongside phylogenetic analyses of 16S rRNA gene sequencing (OQ380604.1). Using bean straw as the best agro-waste, the production of cellulase from this strain was statistically optimized using a response surface methodology, with the maximum activity (13.20 U/mL) achieved at an incubation temperature of 40 °C, a pH of 9, an incubation time of 7 days, and a 2% substrate concentration. The partially purified cellulase (PPC) showed promising activity and stability over a wide range of temperatures (20–90 °C), pH values (3–11), and NaCl concentrations (1–19%), with optimal activity at 50 °C, pH 9.0, and 10% salinity. Under these conditions, the enzyme retained >95% of its activity, thus indicating its extremozyme nature. The kinetics of cellulase showed that it has a V<sub>max</sub> of 20.19 ± 1.88 U/mL and a Km of 0.25 ± 0.07 mM. The immobilized PPC had a relative activity of 69.58 ± 0.13%. In the in vitro microtiter assay, the PPC was found to have a concentration-dependent anti-biofilm activity (up to 85.15 ± 1.60%). Additionally, the fermentative conversion of the hydrolyzed bean straw by <i>Saccharomyces cerevisiae</i> (KM504287.1) amounted to 65.80 ± 0.52% of the theoretical ethanol yield. Overall, for the first time, the present work reports the production of extremozymatic (thermo, alkali-, and halo-stable) cellulase from <i>N. synnemataformans</i> NBRM9. Therefore, this strain is recommended for use as a biotool in many lignocellulosic-based applications operating under harsh conditions.https://www.aimspress.com/article/doi/10.3934/microbiol.2024010?viewType=HTMLcellulasebean strawstatistical optimizationanti-biofilmbioethanol |
| spellingShingle | Mohamed H. El-Sayed Doaa A. Elsayed Abd El-Rahman F. Gomaa <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications AIMS Microbiology cellulase bean straw statistical optimization anti-biofilm bioethanol |
| title | <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications |
| title_full | <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications |
| title_fullStr | <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications |
| title_full_unstemmed | <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications |
| title_short | <i>Nocardiopsis synnemataformans</i> NBRM9, an extremophilic actinomycete producing extremozyme cellulase, using lignocellulosic agro-wastes and its biotechnological applications |
| title_sort | i nocardiopsis synnemataformans i nbrm9 an extremophilic actinomycete producing extremozyme cellulase using lignocellulosic agro wastes and its biotechnological applications |
| topic | cellulase bean straw statistical optimization anti-biofilm bioethanol |
| url | https://www.aimspress.com/article/doi/10.3934/microbiol.2024010?viewType=HTML |
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