Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling
Abstract αL-rhamnosidase (EC 3.2.1.40) has been widely used in food processing and pharmaceutical preparation. The recombinant α-L-rhamnosidase N12-Rha from Aspergillus niger JMU-TS528 had significantly higher catalytic activity on α-1,6 glycosidic bond than α-1,2 glycosidic bond, and had no activit...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
SpringerOpen
2022-11-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-022-01489-5 |
| _version_ | 1798018417003855872 |
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| author | Bo Yu Shiyu Luo Yuhan Ding Zijie Gong Ting Nie |
| author_facet | Bo Yu Shiyu Luo Yuhan Ding Zijie Gong Ting Nie |
| author_sort | Bo Yu |
| collection | DOAJ |
| description | Abstract αL-rhamnosidase (EC 3.2.1.40) has been widely used in food processing and pharmaceutical preparation. The recombinant α-L-rhamnosidase N12-Rha from Aspergillus niger JMU-TS528 had significantly higher catalytic activity on α-1,6 glycosidic bond than α-1,2 glycosidic bond, and had no activity on α-1,3 glycosidic bond. The activities of hydrolyzed hesperidin and naringin were 7240 U/mL and 945 U/mL, respectively, which are 10.63 times that of native α-L-rhamnosidase. The activity could maintain more than 80% at pH 3–6 and 40–60℃. Quantum chemistry calculations showed that charge difference of the C-O atoms of the α-1,2, α-1,3 and α-1,6 bonds indicated that α-1,6 bond is most easily broken and α-1,3 bond is the most stable. Molecular dynamics simulations revealed that the key residue Trp359 that may affect substrate specificity and the main catalytic sites of N12-Rha are located in the (α/α)6-barrel domain. |
| first_indexed | 2024-04-11T16:23:50Z |
| format | Article |
| id | doaj.art-d0b53258553842c4b09c9249d57265f9 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-04-11T16:23:50Z |
| publishDate | 2022-11-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-d0b53258553842c4b09c9249d57265f92022-12-22T04:14:16ZengSpringerOpenAMB Express2191-08552022-11-0112111410.1186/s13568-022-01489-5Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modellingBo Yu0Shiyu Luo1Yuhan Ding2Zijie Gong3Ting Nie4Jiangxi-OAI Joint Research Institute, Nanchang UniversityCollege of Chemistry, Nanchang UniversityMedical College of Dalian UniversityCollege of Chemistry, Nanchang UniversityJiangxi-OAI Joint Research Institute, Nanchang UniversityAbstract αL-rhamnosidase (EC 3.2.1.40) has been widely used in food processing and pharmaceutical preparation. The recombinant α-L-rhamnosidase N12-Rha from Aspergillus niger JMU-TS528 had significantly higher catalytic activity on α-1,6 glycosidic bond than α-1,2 glycosidic bond, and had no activity on α-1,3 glycosidic bond. The activities of hydrolyzed hesperidin and naringin were 7240 U/mL and 945 U/mL, respectively, which are 10.63 times that of native α-L-rhamnosidase. The activity could maintain more than 80% at pH 3–6 and 40–60℃. Quantum chemistry calculations showed that charge difference of the C-O atoms of the α-1,2, α-1,3 and α-1,6 bonds indicated that α-1,6 bond is most easily broken and α-1,3 bond is the most stable. Molecular dynamics simulations revealed that the key residue Trp359 that may affect substrate specificity and the main catalytic sites of N12-Rha are located in the (α/α)6-barrel domain.https://doi.org/10.1186/s13568-022-01489-5α-L-rhamnosidaseCatalytic activityCharge difference |
| spellingShingle | Bo Yu Shiyu Luo Yuhan Ding Zijie Gong Ting Nie Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling AMB Express α-L-rhamnosidase Catalytic activity Charge difference |
| title | Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling |
| title_full | Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling |
| title_fullStr | Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling |
| title_full_unstemmed | Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling |
| title_short | Insights into glycosidic bond specificity of an engineered selective α-L-rhamnosidase N12-Rha via activity assays and molecular modelling |
| title_sort | insights into glycosidic bond specificity of an engineered selective α l rhamnosidase n12 rha via activity assays and molecular modelling |
| topic | α-L-rhamnosidase Catalytic activity Charge difference |
| url | https://doi.org/10.1186/s13568-022-01489-5 |
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