Preparation and Characterization of Chitinase from Marine Bacteria <i>Aeromonas</i> sp. YS-54

CHEN Sunan; LENG Kailiang; YAN Mingyan; LI Yinping; YU Yuan

doi:10.13386/j.issn1002-0306.2023090122

Volume 45 Issue 16

Aug. 2024

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Science and Technology of Food Industry > 2024 > 45(16): 182-190. > DOI: 10.13386/j.issn1002-0306.2023090122

CHEN Sunan, LENG Kailiang, YAN Mingyan, et al. Preparation and Characterization of Chitinase from Marine Bacteria Aeromonas sp. YS-54[J]. Science and Technology of Food Industry, 2024, 45(16): 182−190. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023090122.

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Preparation and Characterization of Chitinase from Marine Bacteria Aeromonas sp. YS-54

1.
College of Marine Science and Biological Engineering, Qingdao University of Science and Technology,Qingdao 266042, China
2.
Key Laboratory of Sustainable Development of Polar Fishery, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China

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Received Date: September 12, 2023
Available Online: June 19, 2024

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Abstract

Abstract

To achieve efficient degradation of crustacean raw materials and eco-preparation of N-acetyl-oligosaccharides, YS-54 strain screened from offshore soil in Qingdao was used for chitinase preparation in the present study. Chitinase was prepared through 60% ammonium sulfate precipitation and Sephadex G-100 gel filtration chromatography. Enzymatic properties such as optimal temperature, optimal pH, and substrate specificity were characterized. Mass spectrometry was used to determine the polymerization degree of enzymatic products. The 16S rRNA identification result showed that YS-54 strain belonged to the Aeromonas species. Chitinase from Aeromonas sp. YS-54 showed a specific activity of 23.44 U/mg. The molecular weights of chitinase were 63 and 75 kDa as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Chitinase was stable under the condition of 40 ℃ and pH5.0. The enzymatic activity was significantly activated by Ba²⁺, Co²⁺, Mg²⁺ and TritonX-100 (P<0.05), while inhibited by Fe³⁺, Cu²⁺ and SDS. The specific activity of chitinase toward α-chitin was 7.99 U/mg, 34.09% of that toward colloidal chitin. The polymerization degree of enzymatic hydrolysis products from colloidal chitin and α-chitin were 1~4 and 1~3, respectively. Chitinase from Aeromonas sp. YS-54 was stable in broad range of temperature and pH, meanwhile showed high catalytic efficiency toward α-chitin. The hydrolysis characteristics of chitinase from Aeromonas sp. YS-54 would provide technical support for the comprehensive utilization of crustacean waste.
- chitinase,
- enzymatic property,
- substrate specificity,
- N-acetyl-oligosaccharides,
- marine bacteria Aeromonas sp.

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