Production of Structurally Defined Chito-Oligosaccharides with a Single N-Acetylation at Their Reducing End Using a Newly Discovered Chitinase from Paenibacillus pabuli

Jing Li, Damao Wang, Shu Chieh Chang, Pi Hui Liang, Vaibhav Srivastava, Shih Yun Guu, Jiun Jie Shie, Kay Hooi Khoo, Vincent Bulone, Yves S.Y. Hsieh

Research output: Contribution to journalArticlepeer-review

Abstract

Partially acetylated chito-oligosaccharides (paCOSs) are bioactive compounds with potential medical applications. Their biological activities are largely dependent on their structural properties, in particular their degree of polymerization (DP) and the position of the acetyl groups along the glycan chain. The production of structurally defined paCOSs in a purified form is highly desirable to better understand the structure/bioactivity relationship of these oligosaccharides. Here, we describe a newly discovered chitinase from Paenibacillus pabuli (PpChi) and demonstrate by mass spectrometry that it essentially produces paCOSs with a DP of three and four that carry a single N-acetylation at their reducing end. We propose that this specific composition of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) residues, as in GlcN(n)GlcNAc1, is due to a subsite specificity toward GlcN residues at the -2, -3, and -4 positions of the partially acetylated chitosan substrates. In addition, the enzyme is stable, as evidenced by its long shelf life, and active over a large temperature range, which is of high interest for potential use in industrial processes. It exhibits a kcat of 67.2 s-1 on partially acetylated chitosan substrates. When PpChi was used in combination with a recently discovered fungal auxilary activity (AA11) oxidase, a sixfold increase in the release of oligosaccharides from the lobster shell was measured. PpChi represents an attractive biocatalyst for the green production of highly valuable paCOSs with a well-defined structure and the expansion of the relatively small library of chito-oligosaccharides currently available.

Original languageEnglish
Pages (from-to)3371-3379
JournalJournal of Agricultural and Food Chemistry
Volume69
Issue number11
DOIs
Publication statusPublished - Mar 24 2021

Keywords

  • Chitin
  • Chitinase
  • Chito-oligosaccharides
  • Chitosan

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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