Expression of recombinant antioxidative peptides from hairy basil ocimum basilicum L. Seed waste in escherichia coli

¡ÒÃáÊ´§ÍÍ¡¢Í§ÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì·ÕèÁÕÄ·¸ÔìµéÒ¹ÍÍ¡«Ôപѹ¨Ò¡¡Ò¡àÁÅç´áÁ§ÅÑ¡ ocimum basilicum L. ã¹ escherichia coli

Name: Apapun Kongcharoen

keyword: Basil

; Escherichia coli

; Antioxidants

; Recombinant proteins

Abstract: Antioxidant activities in plants were widely investigated due to reduction of free radicals which are risks of some diseases. Previously report revealed a potential antioxidative peptide from seed waste of hairy basil. The peptide with amino acid sequence as QTFQYSRGWTN was selected for synthesizing recombinant antioxidant peptide and expressed in Escherichia coli MG1655. This recombinant E. coli named as OSW strain containing the DNA fragment encoding sevenÿcopies of the target peptide on pQE-30 Xa expression vector. The OSW strain could express the recombinant OSW peptide in the soluble fraction. The recombinant OSW peptide elution from Ni2+ affinity column wasÿcontaminated with other proteins. The expected 15 kDa OSW peptide band was further extracted from SDS gel, resulted in the loss of more target peptide, was verified by modified dot blot analysis with antiHis-HPR antibody. Moreover, the recombinant OSW peptide expression was verified by qRT-PCR. The elution of recombinant OSW peptide was examined for the DPPH and ABTS radical scavenging activities, and in vitro protective effect on DNA damage induced by hydroxyl radical. The recombinant OSW elution revealed significantly higher antioxidant activities that those of the chemical synthesized one, particularly the DNA damage prevention. However, for further study, this strain should be subjected for optimizing factors involved in recombinant peptide production and additional purification

Abstract: ¾×ªà»ç¹ÊÔè§ÁÕªÕÇÔµ·ÕèÁÕ¡ÒÃÈÖ¡ÉҤسÊÁºÑµÔ¡Òõéҹ͹ØÁÙÅÍÔÊÃÐÍÂèÒ§á¾ÃèËÅÒ à¹×èͧ¨Ò¡¤ÇÒÁÊÒÁÒö㹡ÒÃÂѺÂÑé§Í¹ØÁÙÅÍÔÊÃÐáÅлéͧ¡Ñ¹¡ÒÃà¡Ô´âäµèÒ§æ §Ò¹ÇԨѷÕè¼èÒ¹ÁÒä´éáÊ´§ãËéàË繶֧ķ¸Ôì¡ÒõéÒ¹ÍÍ¡«Ôപѹ¢Í§à¾»ä·´ì¨Ò¡¡Ò¡àÁÅç´áÁ§ÅÑ¡ ྻ䷴ì·ÕèÁÕÅӴѺ¡Ã´ÍÐÁÔâ¹ QTFQYSRGWTN ¶Ù¡¹ÓÁÒÊѧà¤ÃÒÐËì´éÇÂÇÔ¸Õ·Ò§à¤ÁÕ áÅÐãªéà·¤¹Ô¤´ÕàÍç¹àÍÊÒ¼ÊÁà¾×èÍÊÃéÒ§ Escherichia coli ÊÒ¾ѹ¸Øì·ÕèÁÕ¡ÒÃáÊ´§ÍÍ¡¢Í§ÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì´Ñ§¡ÅèÒǨӹǹ 7 ªØ´ º¹àÍç¡à¾ÃʪѹàǤàµÍÃì pQE-30 Xa àÃÕ¡ÇèÒÊÒ¾ѹ¸Øì OSW ¼Å¡Ò÷´Åͧ¾ºÇèÒÃÕ¤ÍÁºÔá¹¹·ì E. coli ÊÒ¾ѹ¸Øì OSW ÁÕ¡ÒÃáÊ´§ÍÍ¡¢Í§à¾»ä·´ì·Õèµéͧ¡Òà â´ÂÍÂÙèã¹ÃÙ»¢Í§â»ÃµÕ¹·ÕèÅÐÅÒÂÍÂÙèã¹à«ÅÅì ÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì¶Ù¡·ÓãËéºÃÔÊØ·¸Ôì´éÇ Ni2+ ¤ÍÅÑÁ¹ì«Öè§Âѧ¾º¡Òû¹à»×é͹¢Í§â»ÃµÕ¹Í×è¹æ ¨Ö§ãªé¡ÒÃʡѴྻ䷴ì·Õèµéͧ¡Òèҡ SDS à¨Å·Õ袹Ҵ 15 ¡ÔâÅ´Òŵѹ à¾×èÍãËéä´éྻ䷴ì·ÕèºÃÔÊØ·¸Ôì «Öè§à¡Ô´¡ÒÃÊÙ­àÊÕÂྻ䷴ì·Õèµéͧ¡ÒúҧÊèǹä»ÃÐËÇèÒ§¡Ãкǹ¡ÒùÕé ÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì¹Õé¶Ù¡¹Ó仵ÃǨÊͺ´éÇ´éÇÂÇÔ¸Õ¡ÒÃÇÔà¤ÃÒÐËìẺ dot blot ¹Í¡¨Ò¡¹ÕéÂѧÁÕ¡ÒõÃǨÊͺÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì´éÇÂÇÔ¸Õ qRT-PCR ÍÕ¡´éÇ áµèÍÂèÒ§äáçµÒÁ OSW ྻ䷴ì·Õèä´é¨Ò¡¡Ò÷ÓãËéºÃÔÊØ·¸Ôì¨Ò¡¤ÍÅÑÁ¹ì ä´é¶Ù¡¹Óä»·´Êͺķ¸Ôì¡Òõéҹ͹ØÁÙÅÍÔÊÃеèÍ DPPH áÅÐ ABTS ÃÇÁ件֧¡Ò÷´Êͺ¡ÒÃÍÍ¡Ä·¸Ôì»éͧ¡Ñ¹¡Òö١·ÓÅÒ¢ͧ´ÕàÍç¹àÍ ¼Å¡Ò÷´ÊͺáÊ´§ãËéàËç¹ÇèÒ ÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì OSW ·Õèä´é¨Ò¡¡Ò÷ÓãËéºÃÔÊØ·¸Ôì¨Ò¡¤ÍÅÑÁ¹ì ÁÕÄ·¸Ôì¡Òõéҹ͹ØÁÙÅÍÔÊÃÐ ABTS ´Õ¡ÇèÒྻ䷴ì·Õèä´é¨Ò¡¡ÒÃÊѧà¤ÃÒÐËì·Ò§à¤ÁÕÍÂèÒ§ÁÕ¹ÑÂÊӤѭ·Ò§Ê¶ÔµÔ·Õè¤ÇÒÁàª×èÍÁÑè¹ 95% áÅÐÊÒÁÒöÍÍ¡Ä·¸Ôì»éͧ¡Ñ¹¡Òö١·ÓÅÒ¢ͧ´ÕàÍç¹àÍä´é´Õ¡ÇèÒàªè¹¡Ñ¹ ´Ñ§¹Ñé¹ÃÕ¤ÍÁºÔá¹¹·ì E. coli ÊÒ¾ѹ¸Øì OSW ´Ñ§¡ÅèÒǹÕé¨Ö§ÊÒÁÒö¹Óä»ÈÖ¡ÉÒµèÍ㹡ÒÃËһѨ¨Ñ·ÕèàËÁÒÐÊÁ¢Í§áÊ´§ÍÍ¡áÅСÒ÷ÓãËéºÃÔÊØ·¸Ôì à¾×èÍ·Õè¨Ðà»ç¹·Ò§àÅ×͡㹡ÒÃãªé¼ÅÔµÃÕ¤ÍÁºÔá¹¹·ìྻ䷴ì·ÕèÁÕ»ÃÐÊÔ·¸ÔÀÒ¾ÁÒ¡ÂÔ觢Öé¹µèÍä»

Chulalongkorn University. Office of Academic Resources

Address: BANGKOK

Email: cuir@car.chula.ac.th

Role: advisor

Name: Ruethairat Boonsombat

Role: advisor

Created: 2015

Modified: 2023-02-15

Issued: 2023-01-25

ÇÔ·ÂÒ¹Ô¾¹¸ì/Thesis

application/pdf

URL: http://cuir.car.chula.ac.th/handle/123456789/76800

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biotechnology

Grantor: Chulalongkorn University

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