Preparation of Chitooligosaccharides with Low Degree of Deacetylation by Recombinant Chitinase from Pyrococcus furiosus, Its Compositions and Structure Analysis

doi:10.7506/spkx1002-6630-201818018

Abstract

Abstract: The chitinase-encoding gene of Pyrococcus furiosus was synthesized and the recombinant enzyme solubly expressed in Escherichia coli. Chitosan with low degree of deacetylation was hydrolyzed by this enzyme and the composition and structure of the resulting chitooligosaccharides were analyzed. Size exclusion high performance liquid chromatography (SE-HPLC) results showed that the relative molecular masses of the hydrolysis products ranged from 1 000 to 5 000. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis revealed that the hydrolysates contained chitooligosaccharides with a degree of polymerization of 2?9 and different degrees of deacetylation. Structural characterization by nuclear magnetic resonance (NMR) indicated that the reducing end of all oligosaccharide fractions was mainly composed of two successive N-acetylglucosamines. In summary, we have prepared chitooligosaccharides with low degree deacetylation having determined reducing end structure using recombinant chitinase from P. furiosus, which can lay the foundation for the study of the structural-activity relationship of chitooligosaccharides with complex structures.

Key words: Pyrococcus furiosus, chitinase, chitooligosaccharides, size exclusion high performance liquid chromatography (SE-HPLC), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), nuclear magnetic resonance (NMR)

CLC Number:

TS245.9

CHENG Gong, JIAO Siming, REN Lishi, FENG Cui, KANG Jiyao, DU Yuguang. Preparation of Chitooligosaccharides with Low Degree of Deacetylation by Recombinant Chitinase from Pyrococcus furiosus, Its Compositions and Structure Analysis[J]. FOOD SCIENCE, 2018, 39(18): 115-119.

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Preparation of Chitooligosaccharides with Low Degree of Deacetylation by Recombinant Chitinase from Pyrococcus furiosus, Its Compositions and Structure Analysis

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