| dc.creator | Ping,Ruo | |
| dc.creator | Sin Teoh,Leong | |
| dc.creator | Kwai Chan,Mooi | |
| dc.creator | Miswan,Noorizan | |
| dc.creator | Yin Khoo,Boon | |
| dc.date | 2016-05-01 | |
| dc.date.accessioned | 2019-05-03T12:45:23Z | |
| dc.date.available | 2019-05-03T12:45:23Z | |
| dc.identifier | https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000300001 | |
| dc.identifier.uri | http://revistaschilenas.uchile.cl/handle/2250/85539 | |
| dc.description | Background: Human is an essential cellular enzyme that 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 that MutS 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. | |
| dc.format | text/html | |
| dc.language | en | |
| dc.publisher | Pontificia Universidad Católica de Valparaíso | |
| dc.relation | 10.1016/j.ejbt.2016.01.007 | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.source | Electronic Journal of Biotechnology v.19 n.3 2016 | |
| dc.subject | Cell density | |
| dc.subject | Pichia pastoris | |
| dc.subject | Total protein concentration | |
| dc.subject | Recombinant protein expression | |
| dc.title | Comparing the expression of human DNA topoisomerase I in KM71H and X33 strains of Pichia pastoris | |
