Abstract
Pyruvate dehydrogenase multienzyme complex was purified from Bacillus caldolyticus. The complex was composed of four polypeptides with molecular masses of 39.8, 41.7, 53.7, and 57.5kDa estimated by SDS-PAGE and they were presumed to be pyruvate decarboxylase (E1, dimeric), lipoate acetyltransferase (E2), and lipoamide dehydrogenase (E3) on the analogy of those from Bacillus stearothermophilus. E1 and E3 were stable at pH 5.7-10.2 and 4.5-11.3, respectively. Halves of E1 and E3 activity were abolished by incubation for 30min at 65°C and 85°C, respectively. Loss of overall activity was principally due to inactivation of E1. Structural changes in the complex incubated at high temperature were studied by fluorescence spectroscopy. The results suggested that the thermal denaturation of the complex proceeded through at least two different steps: inactivations of E1 and E3, and the former process is accompanied by a reduction of the complex size.
| Original language | English |
|---|---|
| Pages (from-to) | 1062-1066 |
| Number of pages | 5 |
| Journal | Bioscience, Biotechnology, and Biochemistry |
| Volume | 57 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - Jan 1 1993 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Analytical Chemistry
- Biochemistry
- Applied Microbiology and Biotechnology
- Molecular Biology
- Organic Chemistry
Cite this
Purification of Pyruvate Dehydrogenase Complex from an Extreme Thermophile, Bacillus Caldolyticus, and Its Thermal Stability. / Hiromasa, Yasuaki; Aso, Yoichi; Yamashita, Shoji.
In: Bioscience, Biotechnology, and Biochemistry, Vol. 57, No. 7, 01.01.1993, p. 1062-1066.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification of Pyruvate Dehydrogenase Complex from an Extreme Thermophile, Bacillus Caldolyticus, and Its Thermal Stability
AU - Hiromasa, Yasuaki
AU - Aso, Yoichi
AU - Yamashita, Shoji
PY - 1993/1/1
Y1 - 1993/1/1
N2 - Pyruvate dehydrogenase multienzyme complex was purified from Bacillus caldolyticus. The complex was composed of four polypeptides with molecular masses of 39.8, 41.7, 53.7, and 57.5kDa estimated by SDS-PAGE and they were presumed to be pyruvate decarboxylase (E1, dimeric), lipoate acetyltransferase (E2), and lipoamide dehydrogenase (E3) on the analogy of those from Bacillus stearothermophilus. E1 and E3 were stable at pH 5.7-10.2 and 4.5-11.3, respectively. Halves of E1 and E3 activity were abolished by incubation for 30min at 65°C and 85°C, respectively. Loss of overall activity was principally due to inactivation of E1. Structural changes in the complex incubated at high temperature were studied by fluorescence spectroscopy. The results suggested that the thermal denaturation of the complex proceeded through at least two different steps: inactivations of E1 and E3, and the former process is accompanied by a reduction of the complex size.
AB - Pyruvate dehydrogenase multienzyme complex was purified from Bacillus caldolyticus. The complex was composed of four polypeptides with molecular masses of 39.8, 41.7, 53.7, and 57.5kDa estimated by SDS-PAGE and they were presumed to be pyruvate decarboxylase (E1, dimeric), lipoate acetyltransferase (E2), and lipoamide dehydrogenase (E3) on the analogy of those from Bacillus stearothermophilus. E1 and E3 were stable at pH 5.7-10.2 and 4.5-11.3, respectively. Halves of E1 and E3 activity were abolished by incubation for 30min at 65°C and 85°C, respectively. Loss of overall activity was principally due to inactivation of E1. Structural changes in the complex incubated at high temperature were studied by fluorescence spectroscopy. The results suggested that the thermal denaturation of the complex proceeded through at least two different steps: inactivations of E1 and E3, and the former process is accompanied by a reduction of the complex size.
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UR - http://www.scopus.com/inward/citedby.url?scp=0010425590&partnerID=8YFLogxK
U2 - 10.1271/bbb.57.1062
DO - 10.1271/bbb.57.1062
M3 - Article
AN - SCOPUS:0010425590
VL - 57
SP - 1062
EP - 1066
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 7
ER -