Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star                         
                        ™

Ihsan Wiratama; Heri Hermansyah; Anondho Wijarnako; Amarila Malik; Raditya Imamul Khalid; Muhamad Sahlan

doi:10.1063/1.5096732

Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™

Ihsan Wiratama, Heri Hermansyah, Anondho Wijarnako, Amarila Malik, Raditya Imamul Khalid, Muhamad Sahlan

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

Abstract

Cancer is a deadly disease so that the medicinal treatment constantly developed. Apoptin is a protein molecule that has potential to be used as a cancer drug because of its activity to induce cell death selectively to the cancer cells only. Cloning apoptin has been successfully performed by amplify gene using PCR with 12-histidine and 8-arginine to be added at C-terminal then ligated into plasmid pOXGW with Gateway system, and then expressed in Bacillus subtilis 168. Plasmids with pOXGW - apop - 12His8Arg can be expressed in B. subtilis. In this study, Bacillus subtilis carrying plasmid was produced with variations of xylose as substrate trigger on liquid medium and as a comparison, Escherichia coli Bl21 Star ^™ transformed with a plasmid pOGW - apop - 12His and then performed for purification of apoptin. The results showed that the recombinant apoptin obtain from B. subtilis 168 compared to Escherichia coli Bl21 Star ^™ is slightly higher, i.e. 568μg/ml and 421μg/ml, respectively.

Original language	English
Title of host publication	3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices
Subtitle of host publication	Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018
Editors	Praswasti P.D.K. Wulan, Misri Gozan, Sotya Astutiningsih, Ghiska Ramahdita, Radon Dhelika, Prasetyanugraheni Kreshanti
Publisher	American Institute of Physics Inc.
ISBN (Electronic)	9780735418226
DOIs	https://doi.org/10.1063/1.5096732
Publication status	Published - 9 Apr 2019
Event	3rd International Symposium of Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2018 - Jakarta, Indonesia Duration: 6 Aug 2018 → 8 Aug 2018

Publication series

Name	AIP Conference Proceedings
Volume	2092
ISSN (Print)	0094-243X
ISSN (Electronic)	1551-7616

Conference

Conference	3rd International Symposium of Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2018
Country/Territory	Indonesia
City	Jakarta
Period	6/08/18 → 8/08/18

Keywords

Apoptin
Bacillus subtilis 168
Escherichia coli Bl21 Star
plasmid pOXGW - apop - 12His8Arg
POGW apop 12His
xylose

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1063/1.5096732

Cite this

Wiratama, I., Hermansyah, H., Wijarnako, A., Malik, A., Khalid, R. I., & Sahlan, M. (2019). Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™. In P. P. D. K. Wulan, M. Gozan, S. Astutiningsih, G. Ramahdita, R. Dhelika, & P. Kreshanti (Eds.), 3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices: Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018 Article 030028 (AIP Conference Proceedings; Vol. 2092). American Institute of Physics Inc.. https://doi.org/10.1063/1.5096732

Wiratama, Ihsan ; Hermansyah, Heri ; Wijarnako, Anondho et al. / Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™. 3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices: Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018. editor / Praswasti P.D.K. Wulan ; Misri Gozan ; Sotya Astutiningsih ; Ghiska Ramahdita ; Radon Dhelika ; Prasetyanugraheni Kreshanti. American Institute of Physics Inc., 2019. (AIP Conference Proceedings).

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title = "Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star {\texttrademark}",

abstract = " Cancer is a deadly disease so that the medicinal treatment constantly developed. Apoptin is a protein molecule that has potential to be used as a cancer drug because of its activity to induce cell death selectively to the cancer cells only. Cloning apoptin has been successfully performed by amplify gene using PCR with 12-histidine and 8-arginine to be added at C-terminal then ligated into plasmid pOXGW with Gateway system, and then expressed in Bacillus subtilis 168. Plasmids with pOXGW - apop - 12His8Arg can be expressed in B. subtilis. In this study, Bacillus subtilis carrying plasmid was produced with variations of xylose as substrate trigger on liquid medium and as a comparison, Escherichia coli Bl21 Star {\texttrademark} transformed with a plasmid pOGW - apop - 12His and then performed for purification of apoptin. The results showed that the recombinant apoptin obtain from B. subtilis 168 compared to Escherichia coli Bl21 Star {\texttrademark} is slightly higher, i.e. 568μg/ml and 421μg/ml, respectively. ",

keywords = "Apoptin, Bacillus subtilis 168, Escherichia coli Bl21 Star, plasmid pOXGW - apop - 12His8Arg, POGW apop 12His, xylose",

author = "Ihsan Wiratama and Heri Hermansyah and Anondho Wijarnako and Amarila Malik and Khalid, {Raditya Imamul} and Muhamad Sahlan",

note = "Publisher Copyright: {\textcopyright} 2019 Author(s).; 3rd International Symposium of Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2018 ; Conference date: 06-08-2018 Through 08-08-2018",

year = "2019",

month = apr,

day = "9",

doi = "10.1063/1.5096732",

language = "English",

series = "AIP Conference Proceedings",

publisher = "American Institute of Physics Inc.",

editor = "Wulan, {Praswasti P.D.K.} and Misri Gozan and Sotya Astutiningsih and Ghiska Ramahdita and Radon Dhelika and Prasetyanugraheni Kreshanti",

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Wiratama, I, Hermansyah, H, Wijarnako, A, Malik, A, Khalid, RI & Sahlan, M 2019, Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™. in PPDK Wulan, M Gozan, S Astutiningsih, G Ramahdita, R Dhelika & P Kreshanti (eds), 3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices: Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018., 030028, AIP Conference Proceedings, vol. 2092, American Institute of Physics Inc., 3rd International Symposium of Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2018, Jakarta, Indonesia, 6/08/18. https://doi.org/10.1063/1.5096732

Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™. / Wiratama, Ihsan; Hermansyah, Heri; Wijarnako, Anondho et al.
3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices: Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018. ed. / Praswasti P.D.K. Wulan; Misri Gozan; Sotya Astutiningsih; Ghiska Ramahdita; Radon Dhelika; Prasetyanugraheni Kreshanti. American Institute of Physics Inc., 2019. 030028 (AIP Conference Proceedings; Vol. 2092).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

TY - GEN

T1 - Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ™

AU - Wiratama, Ihsan

AU - Hermansyah, Heri

AU - Wijarnako, Anondho

AU - Malik, Amarila

AU - Khalid, Raditya Imamul

AU - Sahlan, Muhamad

PY - 2019/4/9

Y1 - 2019/4/9

N2 - Cancer is a deadly disease so that the medicinal treatment constantly developed. Apoptin is a protein molecule that has potential to be used as a cancer drug because of its activity to induce cell death selectively to the cancer cells only. Cloning apoptin has been successfully performed by amplify gene using PCR with 12-histidine and 8-arginine to be added at C-terminal then ligated into plasmid pOXGW with Gateway system, and then expressed in Bacillus subtilis 168. Plasmids with pOXGW - apop - 12His8Arg can be expressed in B. subtilis. In this study, Bacillus subtilis carrying plasmid was produced with variations of xylose as substrate trigger on liquid medium and as a comparison, Escherichia coli Bl21 Star ™ transformed with a plasmid pOGW - apop - 12His and then performed for purification of apoptin. The results showed that the recombinant apoptin obtain from B. subtilis 168 compared to Escherichia coli Bl21 Star ™ is slightly higher, i.e. 568μg/ml and 421μg/ml, respectively.

AB - Cancer is a deadly disease so that the medicinal treatment constantly developed. Apoptin is a protein molecule that has potential to be used as a cancer drug because of its activity to induce cell death selectively to the cancer cells only. Cloning apoptin has been successfully performed by amplify gene using PCR with 12-histidine and 8-arginine to be added at C-terminal then ligated into plasmid pOXGW with Gateway system, and then expressed in Bacillus subtilis 168. Plasmids with pOXGW - apop - 12His8Arg can be expressed in B. subtilis. In this study, Bacillus subtilis carrying plasmid was produced with variations of xylose as substrate trigger on liquid medium and as a comparison, Escherichia coli Bl21 Star ™ transformed with a plasmid pOGW - apop - 12His and then performed for purification of apoptin. The results showed that the recombinant apoptin obtain from B. subtilis 168 compared to Escherichia coli Bl21 Star ™ is slightly higher, i.e. 568μg/ml and 421μg/ml, respectively.

KW - Apoptin

KW - Bacillus subtilis 168

KW - Escherichia coli Bl21 Star

KW - plasmid pOXGW - apop - 12His8Arg

KW - POGW apop 12His

KW - xylose

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BT - 3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices

A2 - Wulan, Praswasti P.D.K.

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Wiratama I, Hermansyah H, Wijarnako A, Malik A, Khalid RI, Sahlan M. Purification of recombinant protein apoptin from two cell host Bacillus subtilis 168 and Escherichia coli Bl21 Star ^™. In Wulan PPDK, Gozan M, Astutiningsih S, Ramahdita G, Dhelika R, Kreshanti P, editors, 3rd Biomedical Engineering''s Recent Progress in Biomaterials, Drugs Development, and Medical Devices: Proceedings of the International Symposium of Biomedical Engineering, ISBE 2018. American Institute of Physics Inc. 2019. 030028. (AIP Conference Proceedings). doi: 10.1063/1.5096732