Abstract
Korean mistletoe lectins (KMLs) possess many biological activities, including anti-tumor, apoptosis-inducing, anti-metastatic, and anti-angiogenic activities. Recombinant KML-1A (rKML-1A) has been previously expressed in soluble form in Escherichia coli cells. However, the expression of rKML-1B in soluble form remains to be accomplished. In this study, we describe the production, purification, and characterization of recombinant KML-1B by using a baculovirus expression system, which employs silkworm larvae and pupae as hosts. Approximately 495 and 702 µg/mL of a 36-kDa rKML-1B protein were produced without a signal peptide by silkworm larvae and pupae, respectively. Treatment of the recombinant protein with N- or O-deglycosylase led to a decrease in its molecular mass, indicating the N-and/or O-glycosylation of rKML-1B. Purified rKML-1B displayed radical scavenging activities toward 1,1-diphenyl-2-picrylhydrazyl and 2,2′-amino-di-[3-ethylbenzthiazoline sulfonate], with a half maximal inhibitory concentration (IC50) of approximately 95 µg/mL. This activity was discovered to be stable at 65°C. To our knowledge, this is the first report detailing the biological activity of plant-derived, pure rKML-1B.
| Original language | English |
|---|---|
| Pages (from-to) | 515-522 |
| Number of pages | 8 |
| Journal | Biotechnology and Bioprocess Engineering |
| Volume | 20 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Jun 25 2015 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
- Biomedical Engineering
Cite this
Expression of recombinant viscum album coloratum lectin B-chain in the silkworm expression system and evaluation of antioxidant activity. / Hong, Sun Mee; Choi, Ji Hyun; Jo, Sun Jung; Song, Seong Kyu; Lee, Man; Kusakabe, Takahiro.
In: Biotechnology and Bioprocess Engineering, Vol. 20, No. 3, 25.06.2015, p. 515-522.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expression of recombinant viscum album coloratum lectin B-chain in the silkworm expression system and evaluation of antioxidant activity
AU - Hong, Sun Mee
AU - Choi, Ji Hyun
AU - Jo, Sun Jung
AU - Song, Seong Kyu
AU - Lee, Man
AU - Kusakabe, Takahiro
PY - 2015/6/25
Y1 - 2015/6/25
N2 - Korean mistletoe lectins (KMLs) possess many biological activities, including anti-tumor, apoptosis-inducing, anti-metastatic, and anti-angiogenic activities. Recombinant KML-1A (rKML-1A) has been previously expressed in soluble form in Escherichia coli cells. However, the expression of rKML-1B in soluble form remains to be accomplished. In this study, we describe the production, purification, and characterization of recombinant KML-1B by using a baculovirus expression system, which employs silkworm larvae and pupae as hosts. Approximately 495 and 702 µg/mL of a 36-kDa rKML-1B protein were produced without a signal peptide by silkworm larvae and pupae, respectively. Treatment of the recombinant protein with N- or O-deglycosylase led to a decrease in its molecular mass, indicating the N-and/or O-glycosylation of rKML-1B. Purified rKML-1B displayed radical scavenging activities toward 1,1-diphenyl-2-picrylhydrazyl and 2,2′-amino-di-[3-ethylbenzthiazoline sulfonate], with a half maximal inhibitory concentration (IC50) of approximately 95 µg/mL. This activity was discovered to be stable at 65°C. To our knowledge, this is the first report detailing the biological activity of plant-derived, pure rKML-1B.
AB - Korean mistletoe lectins (KMLs) possess many biological activities, including anti-tumor, apoptosis-inducing, anti-metastatic, and anti-angiogenic activities. Recombinant KML-1A (rKML-1A) has been previously expressed in soluble form in Escherichia coli cells. However, the expression of rKML-1B in soluble form remains to be accomplished. In this study, we describe the production, purification, and characterization of recombinant KML-1B by using a baculovirus expression system, which employs silkworm larvae and pupae as hosts. Approximately 495 and 702 µg/mL of a 36-kDa rKML-1B protein were produced without a signal peptide by silkworm larvae and pupae, respectively. Treatment of the recombinant protein with N- or O-deglycosylase led to a decrease in its molecular mass, indicating the N-and/or O-glycosylation of rKML-1B. Purified rKML-1B displayed radical scavenging activities toward 1,1-diphenyl-2-picrylhydrazyl and 2,2′-amino-di-[3-ethylbenzthiazoline sulfonate], with a half maximal inhibitory concentration (IC50) of approximately 95 µg/mL. This activity was discovered to be stable at 65°C. To our knowledge, this is the first report detailing the biological activity of plant-derived, pure rKML-1B.
UR - http://www.scopus.com/inward/record.url?scp=84937945926&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84937945926&partnerID=8YFLogxK
U2 - 10.1007/s12257-014-0806-x
DO - 10.1007/s12257-014-0806-x
M3 - Article
AN - SCOPUS:84937945926
VL - 20
SP - 515
EP - 522
JO - Biotechnology and Bioprocess Engineering
JF - Biotechnology and Bioprocess Engineering
SN - 1226-8372
IS - 3
ER -