Glutathione peroxidase-like activity of 33 kDa trypsin inhibitor from roots of sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57')

Wen C. Hou, Hsien Jung Chen, Chuan Hsiao Han, Ching Yang Lin, Yaw Huei Lin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The hydrogen peroxide, glutathione (GSH), 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS) were used for spectrophotometric assay of commercial glutathione peroxidase (GPx) activity at 570nm. A positive correlation (r2=0.998) was found between level of GPx activity and the absorbance changes (%). The 33kDa trypsin inhibitor (TI) was purified from the storage roots of sweet potato (SP) (Ipomoea batatas [L.] Lam 'Tainong 57') by trypsin-Sepharose 4B affinity chromatography and preparative acrylamide gel electrophoresis. The first 15 amino acids in the N-ternimal region of 33kDa TI were SSETPVLDINGDEVR, which were identical to that of deduced sequence of sopramin A or B. A positive correlation (r 2=0.979) was found between the amounts of 33kDa TI added and absorbance changes. Absorbance changes of 5.18, 16.35 and 25.83%, respectively, were obtained when 100, 200 and 250μg 33kDa TI were added, which were equal to 0.56, 1.21 and 1.76 GPx units. Using TI and GPx activity stainings, it was confirmed that 33kDa TI exhibited GPx-like activity. The physiological significance of TIs with GPx activities is discussed.

Original languageEnglish
Pages (from-to)1541-1546
Number of pages6
JournalPlant Science
Volume166
Issue number6
DOIs
Publication statusPublished - Jun 2004

Fingerprint

Ipomoea batatas
Trypsin Inhibitors
trypsin inhibitors
sweet potatoes
Glutathione Peroxidase
glutathione peroxidase
absorbance
Methylphenazonium Methosulfate
phenazines
Affinity chromatography
Acrylamide
acrylamides
affinity chromatography
Electrophoresis
bromides
Affinity Chromatography
Sepharose
Trypsin
trypsin
Hydrogen Peroxide

Keywords

  • Ammonium persulfate
  • APS
  • Bis
  • CBG
  • Commassie brilliant blue G-250
  • Glutathione
  • Glutathione peroxidase
  • GPx
  • GSH
  • N,N′-methylenediacrylamide
  • SDS-PAGE
  • Sodium dodecylsulfate-polyacrylamide gel electrophoresis
  • SP
  • Sweet potato
  • TEMED

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Biotechnology

Cite this

Glutathione peroxidase-like activity of 33 kDa trypsin inhibitor from roots of sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57'). / Hou, Wen C.; Chen, Hsien Jung; Han, Chuan Hsiao; Lin, Ching Yang; Lin, Yaw Huei.

In: Plant Science, Vol. 166, No. 6, 06.2004, p. 1541-1546.

Research output: Contribution to journalArticle

Hou, Wen C. ; Chen, Hsien Jung ; Han, Chuan Hsiao ; Lin, Ching Yang ; Lin, Yaw Huei. / Glutathione peroxidase-like activity of 33 kDa trypsin inhibitor from roots of sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57'). In: Plant Science. 2004 ; Vol. 166, No. 6. pp. 1541-1546.
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abstract = "The hydrogen peroxide, glutathione (GSH), 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS) were used for spectrophotometric assay of commercial glutathione peroxidase (GPx) activity at 570nm. A positive correlation (r2=0.998) was found between level of GPx activity and the absorbance changes ({\%}). The 33kDa trypsin inhibitor (TI) was purified from the storage roots of sweet potato (SP) (Ipomoea batatas [L.] Lam 'Tainong 57') by trypsin-Sepharose 4B affinity chromatography and preparative acrylamide gel electrophoresis. The first 15 amino acids in the N-ternimal region of 33kDa TI were SSETPVLDINGDEVR, which were identical to that of deduced sequence of sopramin A or B. A positive correlation (r 2=0.979) was found between the amounts of 33kDa TI added and absorbance changes. Absorbance changes of 5.18, 16.35 and 25.83{\%}, respectively, were obtained when 100, 200 and 250μg 33kDa TI were added, which were equal to 0.56, 1.21 and 1.76 GPx units. Using TI and GPx activity stainings, it was confirmed that 33kDa TI exhibited GPx-like activity. The physiological significance of TIs with GPx activities is discussed.",
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T1 - Glutathione peroxidase-like activity of 33 kDa trypsin inhibitor from roots of sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57')

AU - Hou, Wen C.

AU - Chen, Hsien Jung

AU - Han, Chuan Hsiao

AU - Lin, Ching Yang

AU - Lin, Yaw Huei

PY - 2004/6

Y1 - 2004/6

N2 - The hydrogen peroxide, glutathione (GSH), 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS) were used for spectrophotometric assay of commercial glutathione peroxidase (GPx) activity at 570nm. A positive correlation (r2=0.998) was found between level of GPx activity and the absorbance changes (%). The 33kDa trypsin inhibitor (TI) was purified from the storage roots of sweet potato (SP) (Ipomoea batatas [L.] Lam 'Tainong 57') by trypsin-Sepharose 4B affinity chromatography and preparative acrylamide gel electrophoresis. The first 15 amino acids in the N-ternimal region of 33kDa TI were SSETPVLDINGDEVR, which were identical to that of deduced sequence of sopramin A or B. A positive correlation (r 2=0.979) was found between the amounts of 33kDa TI added and absorbance changes. Absorbance changes of 5.18, 16.35 and 25.83%, respectively, were obtained when 100, 200 and 250μg 33kDa TI were added, which were equal to 0.56, 1.21 and 1.76 GPx units. Using TI and GPx activity stainings, it was confirmed that 33kDa TI exhibited GPx-like activity. The physiological significance of TIs with GPx activities is discussed.

AB - The hydrogen peroxide, glutathione (GSH), 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS) were used for spectrophotometric assay of commercial glutathione peroxidase (GPx) activity at 570nm. A positive correlation (r2=0.998) was found between level of GPx activity and the absorbance changes (%). The 33kDa trypsin inhibitor (TI) was purified from the storage roots of sweet potato (SP) (Ipomoea batatas [L.] Lam 'Tainong 57') by trypsin-Sepharose 4B affinity chromatography and preparative acrylamide gel electrophoresis. The first 15 amino acids in the N-ternimal region of 33kDa TI were SSETPVLDINGDEVR, which were identical to that of deduced sequence of sopramin A or B. A positive correlation (r 2=0.979) was found between the amounts of 33kDa TI added and absorbance changes. Absorbance changes of 5.18, 16.35 and 25.83%, respectively, were obtained when 100, 200 and 250μg 33kDa TI were added, which were equal to 0.56, 1.21 and 1.76 GPx units. Using TI and GPx activity stainings, it was confirmed that 33kDa TI exhibited GPx-like activity. The physiological significance of TIs with GPx activities is discussed.

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KW - APS

KW - Bis

KW - CBG

KW - Commassie brilliant blue G-250

KW - Glutathione

KW - Glutathione peroxidase

KW - GPx

KW - GSH

KW - N,N′-methylenediacrylamide

KW - SDS-PAGE

KW - Sodium dodecylsulfate-polyacrylamide gel electrophoresis

KW - SP

KW - Sweet potato

KW - TEMED

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U2 - 10.1016/j.plantsci.2004.02.007

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M3 - Article

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VL - 166

SP - 1541

EP - 1546

JO - Plant Science

JF - Plant Science

SN - 0168-9452

IS - 6

ER -