Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris
Human is an essential cellular enzymethat is found in all human cells. As this enzyme is upregulated in cancer cells exceedingly, it is used as a target for cancer chemotherapeutic drug development. As such, producing the in-house enzyme for the purpose to speed up the search for more cost-effecti...
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| Format: | Article |
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Elsevier
2016
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| Online Access: | http://eprints.usm.my/36835/ |
| _version_ | 1848878023350157312 |
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| author | Ruo, Ping Ang Leong, Sin Teoh Mooi, Kwai Chana Miswan, Noorizan Boon, Yin Khoo |
| author_facet | Ruo, Ping Ang Leong, Sin Teoh Mooi, Kwai Chana Miswan, Noorizan Boon, Yin Khoo |
| author_sort | Ruo, Ping Ang |
| building | USM Institutional Repository |
| collection | Online Access |
| description | Human is an essential cellular enzymethat is found in all human cells. As this enzyme is upregulated
in cancer cells exceedingly, it is used as a target for cancer chemotherapeutic drug development. As such,
producing the in-house enzyme for the purpose to speed up the search for more cost-effective and target
specific hTopoI inhibitors is warranted. This study aims to compare the optimised conditions for the
expression of hTopoI in KM71H (MutS) and X33 (Mut+) strains of Pichia pastoris. P. pastoris transfected with
an hTopoI recombinant vector was used for the optimization of a higher level of hTopoI expression.
Results: In the process, fed-batch cultivation parameters that influence the expression of hTopoI, such as culture
temperature, methanol induction and feeding strategy, were optimised in the transfected KM71H and X33
P. pastoris strains in a shake flask system. The cell density and total protein concentration (protein level) of
transfected P. pastoris were compared to determine the optimum culture conditions for each transfected
P. pastoris strain. A higher hTopoI level was observed in the transfected KM71H culture supernatant
(2.26 ng/mL) when the culture was incubated in the optimum conditions.
Conclusions: This study demonstrated thatMutS strain (KM71H) expressed and secreted a higher level of hTopoI
heterologous protein in the presence of methanol compared to the Mut+ strain; X33 (0.75 ng/mL). However,
other aspects of optimization, such as pH, should also be considered in the future, to obtain the optimum
expression level of hTopoI in P. pastoris. |
| first_indexed | 2025-11-15T17:24:44Z |
| format | Article |
| id | usm-36835 |
| institution | Universiti Sains Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T17:24:44Z |
| publishDate | 2016 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | usm-368352017-09-29T07:12:18Z http://eprints.usm.my/36835/ Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris Ruo, Ping Ang Leong, Sin Teoh Mooi, Kwai Chana Miswan, Noorizan Boon, Yin Khoo R735-854 Medical education. Medical schools. Research T1-995 Technology(General) Human is an essential cellular enzymethat is found in all human cells. As this enzyme is upregulated in cancer cells exceedingly, it is used as a target for cancer chemotherapeutic drug development. As such, producing the in-house enzyme for the purpose to speed up the search for more cost-effective and target specific hTopoI inhibitors is warranted. This study aims to compare the optimised conditions for the expression of hTopoI in KM71H (MutS) and X33 (Mut+) strains of Pichia pastoris. P. pastoris transfected with an hTopoI recombinant vector was used for the optimization of a higher level of hTopoI expression. Results: In the process, fed-batch cultivation parameters that influence the expression of hTopoI, such as culture temperature, methanol induction and feeding strategy, were optimised in the transfected KM71H and X33 P. pastoris strains in a shake flask system. The cell density and total protein concentration (protein level) of transfected P. pastoris were compared to determine the optimum culture conditions for each transfected P. pastoris strain. A higher hTopoI level was observed in the transfected KM71H culture supernatant (2.26 ng/mL) when the culture was incubated in the optimum conditions. Conclusions: This study demonstrated thatMutS strain (KM71H) expressed and secreted a higher level of hTopoI heterologous protein in the presence of methanol compared to the Mut+ strain; X33 (0.75 ng/mL). However, other aspects of optimization, such as pH, should also be considered in the future, to obtain the optimum expression level of hTopoI in P. pastoris. Elsevier 2016-05 Article PeerReviewed Ruo, Ping Ang and Leong, Sin Teoh and Mooi, Kwai Chana and Miswan, Noorizan and Boon, Yin Khoo (2016) Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris. Electronic Journal of Biotechnology, 21. pp. 9-17. ISSN 0717-3458 http://dx.doi.org/10.1016/j.ejbt.2016.01.007 |
| spellingShingle | R735-854 Medical education. Medical schools. Research T1-995 Technology(General) Ruo, Ping Ang Leong, Sin Teoh Mooi, Kwai Chana Miswan, Noorizan Boon, Yin Khoo Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title_full | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title_fullStr | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title_full_unstemmed | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title_short | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris |
| title_sort | comparing the expression of human dna topoisomerase i in km71h and x33 strains of pichia pastoris |
| topic | R735-854 Medical education. Medical schools. Research T1-995 Technology(General) |
| url | http://eprints.usm.my/36835/ http://eprints.usm.my/36835/ |