Crystallographic analysis of Eisenia hydrolysis-enhancing protein using a long wavelength for native-SAD phasing

Xiaomei Sun, Yuxin Ye, Naofumi Sakurai, Koji Kato, Keizo Yuasa, Akihiko Tsuji, Min Yao

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)20-24
Number of pages5
JournalActa Crystallographica Section F: Structural Biology Communications
Volume76
DOIs
Publication statusPublished - Jan 1 2020

Fingerprint

hydrolysis
Hydrolysis
Diffraction
proteins
Wavelength
diffraction
wavelengths
Proteins
Phaeophyta
algae
Algae
Glucosidases
Aplysia
Atoms
Crystals
Biofuels
glucose
crystals
Carrier concentration
atoms

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 journalArticle

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