Abstract
Eisenia hydrolysis-enhancing protein (EHEP), which is a novel protein that has been identified in Aplysia kurodai, protects β-glucosidases from phlorotannin inhibition to facilitate the production of glucose from the laminarin abundant in brown algae. Hence, EHEP has attracted attention for its potential applications in producing biofuel from brown algae. In this study, EHEP was purified from the natural digestive fluid of A. kurodai and was crystallized using the sitting-drop vapor-diffusion method. Native and SAD (single-wavelength anomalous diffraction) data sets were successfully collected at resolutions of 1.20 and 2.48Å using wavelengths of 1.0 and 2.1Å, respectively, from crystals obtained in initial screening. The crystals belonged to space group P212121 and contained one EHEP molecule in the asymmetric unit. All 20 S-atom sites in EHEP were located and the phases were determined by the SAD method using the S atoms in the natural protein as anomalous scatterers (native-SAD). After phase improvement, interpretable electron densities were obtained and 58% of the model was automatically built.
| Original language | English |
|---|---|
| Pages (from-to) | 20-24 |
| Number of pages | 5 |
| Journal | Acta Crystallographica Section F: Structural Biology Communications |
| Volume | 76 |
| DOIs | |
| Publication status | Published - Jan 1 2020 |
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Keywords
- biofuel
- EHEP
- native-SAD
- phlorotannin binding
- solutionless crystal mount
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Genetics
- Condensed Matter Physics
Cite this
Crystallographic analysis of Eisenia hydrolysis-enhancing protein using a long wavelength for native-SAD phasing. / Sun, Xiaomei; Ye, Yuxin; Sakurai, Naofumi; Kato, Koji; Yuasa, Keizo; Tsuji, Akihiko; Yao, Min.
In: Acta Crystallographica Section F: Structural Biology Communications, Vol. 76, 01.01.2020, p. 20-24.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Crystallographic analysis of Eisenia hydrolysis-enhancing protein using a long wavelength for native-SAD phasing
AU - Sun, Xiaomei
AU - Ye, Yuxin
AU - Sakurai, Naofumi
AU - Kato, Koji
AU - Yuasa, Keizo
AU - Tsuji, Akihiko
AU - Yao, Min
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Eisenia hydrolysis-enhancing protein (EHEP), which is a novel protein that has been identified in Aplysia kurodai, protects β-glucosidases from phlorotannin inhibition to facilitate the production of glucose from the laminarin abundant in brown algae. Hence, EHEP has attracted attention for its potential applications in producing biofuel from brown algae. In this study, EHEP was purified from the natural digestive fluid of A. kurodai and was crystallized using the sitting-drop vapor-diffusion method. Native and SAD (single-wavelength anomalous diffraction) data sets were successfully collected at resolutions of 1.20 and 2.48Å using wavelengths of 1.0 and 2.1Å, respectively, from crystals obtained in initial screening. The crystals belonged to space group P212121 and contained one EHEP molecule in the asymmetric unit. All 20 S-atom sites in EHEP were located and the phases were determined by the SAD method using the S atoms in the natural protein as anomalous scatterers (native-SAD). After phase improvement, interpretable electron densities were obtained and 58% of the model was automatically built.
AB - Eisenia hydrolysis-enhancing protein (EHEP), which is a novel protein that has been identified in Aplysia kurodai, protects β-glucosidases from phlorotannin inhibition to facilitate the production of glucose from the laminarin abundant in brown algae. Hence, EHEP has attracted attention for its potential applications in producing biofuel from brown algae. In this study, EHEP was purified from the natural digestive fluid of A. kurodai and was crystallized using the sitting-drop vapor-diffusion method. Native and SAD (single-wavelength anomalous diffraction) data sets were successfully collected at resolutions of 1.20 and 2.48Å using wavelengths of 1.0 and 2.1Å, respectively, from crystals obtained in initial screening. The crystals belonged to space group P212121 and contained one EHEP molecule in the asymmetric unit. All 20 S-atom sites in EHEP were located and the phases were determined by the SAD method using the S atoms in the natural protein as anomalous scatterers (native-SAD). After phase improvement, interpretable electron densities were obtained and 58% of the model was automatically built.
KW - biofuel
KW - EHEP
KW - native-SAD
KW - phlorotannin binding
KW - solutionless crystal mount
UR - http://www.scopus.com/inward/record.url?scp=85077765928&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85077765928&partnerID=8YFLogxK
U2 - 10.1107/S2053230X19016716
DO - 10.1107/S2053230X19016716
M3 - Article
C2 - 31929182
AN - SCOPUS:85077765928
VL - 76
SP - 20
EP - 24
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
SN - 1744-3091
ER -