Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin
<i>Aspergillus fumigatus</i> HBFH5 is a thermophilic fungus which can efficiently degrade lignocellulose and which produces a variety of glycoside hydrolase. In the present study, a novel β-mannanase gene (AfMan5A) was expressed in <i>Pichia pastoris</i> and characterized. Af...
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MDPI AG
2021-02-01
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| author | Xinxi Gu Haiqiang Lu Wei Chen Xiangchen Meng |
| author_facet | Xinxi Gu Haiqiang Lu Wei Chen Xiangchen Meng |
| author_sort | Xinxi Gu |
| collection | DOAJ |
| description | <i>Aspergillus fumigatus</i> HBFH5 is a thermophilic fungus which can efficiently degrade lignocellulose and which produces a variety of glycoside hydrolase. In the present study, a novel β-mannanase gene (AfMan5A) was expressed in <i>Pichia pastoris</i> and characterized. AfMan5A is composed of 373 amino acids residues, and has a calculated molecular weight of 40 kDa. It has been observed that the amino acid sequence of AfMan5A showed 74.4% homology with the ManBK from <i>Aspergillus niger</i>. In addition, the recombined AfMan5A exhibited optimal hydrolytic activity at 60 °C and pH 6.0. It had no activity loss after incubation for 1h at 60 °C, while 65% of the initial activity was observed after 1 h at 70 °C. Additionally, it maintained about 80% of its activity in the pH range from 3.0 to 9.0. When carob bean gum was used as the substrate, the Km and Vmax values of AfMan5A were 0.21 ± 0.05 mg·mL<sup>−1</sup> and 15.22 ± 0.33 U mg<sup>−1</sup>·min<sup>−1</sup>, respectively. AfMan5A and AfSwol showed a strong synergistic interaction on galactomannan degradation, increasing the reduction of the sugars by up to 31%. Therefore, these findings contribute to new strategies for improving the hydrolysis of galactomannan using the enzyme cocktail. |
| first_indexed | 2024-03-09T00:53:52Z |
| format | Article |
| id | doaj.art-c3b93b756d134ee790e1effcb34458b4 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-09T00:53:52Z |
| publishDate | 2021-02-01 |
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| spelling | doaj.art-c3b93b756d134ee790e1effcb34458b42023-12-11T17:01:47ZengMDPI AGCatalysts2073-43442021-02-0111225410.3390/catal11020254Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with SwolleninXinxi Gu0Haiqiang Lu1Wei Chen2Xiangchen Meng3Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaKey Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China<i>Aspergillus fumigatus</i> HBFH5 is a thermophilic fungus which can efficiently degrade lignocellulose and which produces a variety of glycoside hydrolase. In the present study, a novel β-mannanase gene (AfMan5A) was expressed in <i>Pichia pastoris</i> and characterized. AfMan5A is composed of 373 amino acids residues, and has a calculated molecular weight of 40 kDa. It has been observed that the amino acid sequence of AfMan5A showed 74.4% homology with the ManBK from <i>Aspergillus niger</i>. In addition, the recombined AfMan5A exhibited optimal hydrolytic activity at 60 °C and pH 6.0. It had no activity loss after incubation for 1h at 60 °C, while 65% of the initial activity was observed after 1 h at 70 °C. Additionally, it maintained about 80% of its activity in the pH range from 3.0 to 9.0. When carob bean gum was used as the substrate, the Km and Vmax values of AfMan5A were 0.21 ± 0.05 mg·mL<sup>−1</sup> and 15.22 ± 0.33 U mg<sup>−1</sup>·min<sup>−1</sup>, respectively. AfMan5A and AfSwol showed a strong synergistic interaction on galactomannan degradation, increasing the reduction of the sugars by up to 31%. Therefore, these findings contribute to new strategies for improving the hydrolysis of galactomannan using the enzyme cocktail.https://www.mdpi.com/2073-4344/11/2/254mannanaseswolleninsynergisticgalactomannanhydrolysis |
| spellingShingle | Xinxi Gu Haiqiang Lu Wei Chen Xiangchen Meng Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin Catalysts mannanase swollenin synergistic galactomannan hydrolysis |
| title | Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin |
| title_full | Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin |
| title_fullStr | Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin |
| title_full_unstemmed | Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin |
| title_short | Characterization of a Novel Thermophilic Mannanase and Synergistic Hydrolysis of Galactomannan Combined with Swollenin |
| title_sort | characterization of a novel thermophilic mannanase and synergistic hydrolysis of galactomannan combined with swollenin |
| topic | mannanase swollenin synergistic galactomannan hydrolysis |
| url | https://www.mdpi.com/2073-4344/11/2/254 |
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