Observation of in vitro cellulose synthesis by bacterial cellulose synthase with time-resolved small angle X-ray scattering

Hirotaka Tajima, Paavo A. Penttilä, Tomoya Imai*, Kyoko Yamamoto, Yoshiaki Yuguchi

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

3 Citations (Scopus)

Abstract

Cellulose synthase is the enzyme that produces cellulose in the living organisms like plant, and has two functions: polymerizing glucose residues (polymerization) and assembling these polymerized molecules into a crystalline microfibril with a “cellulose I” crystallographic structure (crystallization). Many studies, however, have shown that an in vitro reaction of cellulose synthase produces aggregates of a non-native crystallographic structure “cellulose II” despite the remaining polymerizing activity. This is partial denaturation or loss of crystallization function in cellulose synthase, which needs to be resolved to reconstitute its native activity. To this end, we aimed to clarify the process of cellulose II formation by bacterial cellulose synthase in vitro, using in situ small angle X-ray scattering (SAXS). An increase in scattering specific to synthesis was observed around two distinct regions of q (0.2–0.4 nm −1 and <0.1 nm −1 ) by time-resolved SAXS measurement. The scattering at higher q-region appears prior to lower-q scattering at beginning of the reaction, indicating the existence of smaller primitive aggregations at the initiation stage. This study demonstrates the use of in situ SAXS measurement to decipher the dynamics of biosynthesized cellulose chains, which is a remarkable example of polymer assembly in ambient conditions.

Original languageEnglish
Pages (from-to)765-777
Number of pages13
JournalInternational Journal of Biological Macromolecules
Volume130
DOIs
Publication statusPublished - 1 Jun 2019
MoE publication typeA1 Journal article-refereed

Keywords

  • Cellulose crystallization
  • Cellulose synthase
  • In situ observation
  • Small angle X-ray scattering (SAXS)

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