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Ketudat-Cairns, James R. . Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7. (). King Mongkut's University of Technology North Bangkok. Central Library. : , 2021.
Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7
Name: Istiftakhun Nikmah
Organization :
Suranaree University of Technology. School of Chemistry, Institute of Science and Center of Biomolecular Structure, Function, and Application
Name: Sittiruk Roytrakul
Organization :
National Center for Genetic Engineering and Biotechnology
Name: Fatchiyah Fatchiyah
Organization :
Brawijaya University. Faculty of Mathematics and Natural Science
Name: Ketudat-Cairns, James R.
Organization :
Chulabhorn Research. Laboratory of Biochemistry
Email :
cairns@sut.ac.th
keyword:
Alcohol oxidase 1
ThaSH:
Pichia pastoris
ThaSH:
Anthocyanins
Abstract:
Pichia pastoris is a popular host for recombinant expression of proteins due to
its ability to secrete proteins, grow at high cell density, grow on media with methanol
as a carbon source, and support methanol-induced protein expression with the alcohol
oxidase I (AOX1) promoter. Although the AOX1 protein is localized in the
peroxisome, we have noted secretion to the media upon expression of certain genes. In
this study, two rice serine carboxypeptidase-like (SCPL) acyltransferase genes, i.e.,
OsSCPL2a and OsSCPL7, in pPICZαBNH8 were used to transform into P. pastoris for
expression. Upon induction of expression, a protein of 68 kDa was observed in the
medium of cells expressing OsSCPL7, but not those expressing OsSCPL2a. The
protein was purified by immobilized metal affinity chromatography (IMAC) from the
media, excised from an SDS-PAGE gel, and identified by liquid chromatography and
tandem mass spectrometry (LC/MS/MS) as Pichia AOX1. The purified AOX1 was
found to convert cyanidin-3-O-glucoside (C3G) to compounds that no longer exhibit
absorbance in the 520-530 nm range, but absorb relatively strongly at 360 nm, as
detected in ultra-high-performance liquid chromatography (UHPLC). These results
suggest that AOX1 protein may be released into Pichia medium during expression of
certain proteins, but not other closely related proteins, and suggests that AOXI has the
ability to oxidize complex phenolic alcohols, such as anthocyanins.
King Mongkut's University of Technology North Bangkok. Central Library
Address:
BANGKOK
Email:
library@kmutnb.ac.th
Created:
2021
Modified:
2023-05-31
Issued:
2023-05-31
บทความ/Article
application/pdf
BibliograpyCitation :
In Thai Society for Biotechnology. Thai Society for Biotechnology and International Conference Online (TSB Online 2021) (pp.416-424). Bangkok : Thai Society for Biotechnology
eng
©copyrights King Mongkut's University of Technology North Bangkok
RightsAccess:
ใช้เวลา
0.039446 วินาที
Istiftakhun Nikmah
| Title | Contributor | Type |
|---|---|---|
| Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7
มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าพระนครเหนือ Istiftakhun Nikmah;Sittiruk Roytrakul;Fatchiyah Fatchiyah;Ketudat-Cairns, James R. | บทความ/Article |
Sittiruk Roytrakul
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|---|---|---|
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| Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7
มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าพระนครเหนือ Istiftakhun Nikmah;Sittiruk Roytrakul;Fatchiyah Fatchiyah;Ketudat-Cairns, James R. | บทความ/Article | |
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Fatchiyah Fatchiyah
| Title | Contributor | Type |
|---|---|---|
| Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7
มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าพระนครเหนือ Istiftakhun Nikmah;Sittiruk Roytrakul;Fatchiyah Fatchiyah;Ketudat-Cairns, James R. | บทความ/Article |
Ketudat-Cairns, James R.
| Title | Contributor | Type |
|---|---|---|
| Release of alcohol oxidase I (AOX1) protein from Pichia pastoris during expression of rice serine carboxypeptidase-like 7
มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าพระนครเหนือ Istiftakhun Nikmah;Sittiruk Roytrakul;Fatchiyah Fatchiyah;Ketudat-Cairns, James R. | บทความ/Article |