Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system
Human hexokinase II (HKII) is one of the key enzymes in the glycolytic pathway. It has been postulated that HKII is a potential target for anti-dengue (DENV) drug development, as well as involved in cancer and tumor cell growth. In this work, the human hexokinase II (HKII) gene was cloned into p...
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| Format: | Proceeding Paper |
| Language: | English English |
| Published: |
Association for Computing Machinery
2019
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/79728/ http://irep.iium.edu.my/79728/9/79728_Expression%20and%20Purification%20of%20Soluble_new_complete.pdf http://irep.iium.edu.my/79728/2/79728_Expression%20and%20Purification%20of%20Soluble%20Bacterially-Expressed_scopus.pdf |
| _version_ | 1848788827474231296 |
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| author | Tanbin, Suriyea Ahmad Fuad, Fazia Adyani |
| author_facet | Tanbin, Suriyea Ahmad Fuad, Fazia Adyani |
| author_sort | Tanbin, Suriyea |
| building | IIUM Repository |
| collection | Online Access |
| description | Human hexokinase II (HKII) is one of the key enzymes in the
glycolytic pathway. It has been postulated that HKII is a potential
target for anti-dengue (DENV) drug development, as well as
involved in cancer and tumor cell growth. In this work, the
human hexokinase II (HKII) gene was cloned into pETite N-His
SUMO vector and transformed into the E.coli strain HI-control
10G for the propagation of clones. Two different expression
hosts, E.coli HI-controlTM BL21 (DE3) and BL21 (DE3) pLysS
were used to optimize HKII expression. In order to obtain the
soluble recombinant HKII in a functional form, we optimized
protein expression at three different temperatures; 17°C, 25°C and
37°C, at 24 hours incubation time. The soluble protein was
expressed in the presence of 0.5 mM isopropyl-2-Dthiogalactopyranoside (IPTG) in TB media at 17°C for 24 hrs.
The expressed protein was then purified to homogeneity by a
combination of Immobilized Metal Ion Affinity Chromatography
(IMAC), size exclusion chromatography (SEC) and ion-exchange
chromatography (IEX), resulting in pure bacterially-expressed
HK2. Taken together, this study has successfully produced
soluble bacterially-expressed human HKII that can be utilized for
further therapeutic studies. |
| first_indexed | 2025-11-14T17:47:01Z |
| format | Proceeding Paper |
| id | iium-79728 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-14T17:47:01Z |
| publishDate | 2019 |
| publisher | Association for Computing Machinery |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-797282022-03-17T08:08:19Z http://irep.iium.edu.my/79728/ Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system Tanbin, Suriyea Ahmad Fuad, Fazia Adyani QH Natural history TP248.13 Biotechnology Human hexokinase II (HKII) is one of the key enzymes in the glycolytic pathway. It has been postulated that HKII is a potential target for anti-dengue (DENV) drug development, as well as involved in cancer and tumor cell growth. In this work, the human hexokinase II (HKII) gene was cloned into pETite N-His SUMO vector and transformed into the E.coli strain HI-control 10G for the propagation of clones. Two different expression hosts, E.coli HI-controlTM BL21 (DE3) and BL21 (DE3) pLysS were used to optimize HKII expression. In order to obtain the soluble recombinant HKII in a functional form, we optimized protein expression at three different temperatures; 17°C, 25°C and 37°C, at 24 hours incubation time. The soluble protein was expressed in the presence of 0.5 mM isopropyl-2-Dthiogalactopyranoside (IPTG) in TB media at 17°C for 24 hrs. The expressed protein was then purified to homogeneity by a combination of Immobilized Metal Ion Affinity Chromatography (IMAC), size exclusion chromatography (SEC) and ion-exchange chromatography (IEX), resulting in pure bacterially-expressed HK2. Taken together, this study has successfully produced soluble bacterially-expressed human HKII that can be utilized for further therapeutic studies. Association for Computing Machinery 2019-03-28 Proceeding Paper NonPeerReviewed application/pdf en http://irep.iium.edu.my/79728/9/79728_Expression%20and%20Purification%20of%20Soluble_new_complete.pdf application/pdf en http://irep.iium.edu.my/79728/2/79728_Expression%20and%20Purification%20of%20Soluble%20Bacterially-Expressed_scopus.pdf Tanbin, Suriyea and Ahmad Fuad, Fazia Adyani (2019) Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system. In: "9th International Conference on Biomedical Engineering and Technology, ICBET 2019", 28 - 30 Mar 2019, Meiji UniversityTokyo, Japan. https://dl.acm.org/doi/pdf/10.1145/3326172.3326219 10.1145/3326172.3326219 |
| spellingShingle | QH Natural history TP248.13 Biotechnology Tanbin, Suriyea Ahmad Fuad, Fazia Adyani Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title | Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title_full | Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title_fullStr | Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title_full_unstemmed | Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title_short | Expression and purification of soluble bacterially-expressed human hexokinase II in E.coli system |
| title_sort | expression and purification of soluble bacterially-expressed human hexokinase ii in e.coli system |
| topic | QH Natural history TP248.13 Biotechnology |
| url | http://irep.iium.edu.my/79728/ http://irep.iium.edu.my/79728/ http://irep.iium.edu.my/79728/ http://irep.iium.edu.my/79728/9/79728_Expression%20and%20Purification%20of%20Soluble_new_complete.pdf http://irep.iium.edu.my/79728/2/79728_Expression%20and%20Purification%20of%20Soluble%20Bacterially-Expressed_scopus.pdf |