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中国精品科技期刊2020
谢啟发,薛小旭,黄志发,等. 海洋产几丁质酶厦门加西利亚单胞菌Chi34的筛选、鉴定及酶学性质分析[J]. 食品工业科技,2023,44(12):116−123. doi: 10.13386/j.issn1002-0306.2022090266.
引用本文: 谢啟发,薛小旭,黄志发,等. 海洋产几丁质酶厦门加西利亚单胞菌Chi34的筛选、鉴定及酶学性质分析[J]. 食品工业科技,2023,44(12):116−123. doi: 10.13386/j.issn1002-0306.2022090266.
XIE Qifa, XUE Xiaoxu, HUANG Zhifa, et al. Screening, Identification and Characterization of a Marine Chitinase-Producing Strain Gallaecimonas xiamenensis Chi34[J]. Science and Technology of Food Industry, 2023, 44(12): 116−123. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022090266.
Citation: XIE Qifa, XUE Xiaoxu, HUANG Zhifa, et al. Screening, Identification and Characterization of a Marine Chitinase-Producing Strain Gallaecimonas xiamenensis Chi34[J]. Science and Technology of Food Industry, 2023, 44(12): 116−123. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022090266.

海洋产几丁质酶厦门加西利亚单胞菌Chi34的筛选、鉴定及酶学性质分析

Screening, Identification and Characterization of a Marine Chitinase-Producing Strain Gallaecimonas xiamenensis Chi34

  • 摘要: 目的:海洋是自然界中含几丁质最多的生态系统,孵育了大量可降解利用几丁质的微生物,因此利用海洋微生物几丁质酶降解几丁质,是获得高附加值几丁寡糖的重要方法。本试验以连云港海域海泥为样品,以期筛选获得稳定的产几丁质酶菌株。方法:利用平板筛选法和摇瓶发酵复筛法筛选获得产几丁质酶菌株Chi34,利用形态学分析和16S rDNA序列分析对菌株进行鉴定,同时还研究了温度、pH、金属离子、EDTA及SDS等因素对Chi34几丁质酶的影响,最后通过TLC实验分析了Chi34几丁质酶降解几丁质胶体的产物。结果:菌株Chi34鉴定为厦门加西利亚单胞菌(Gallaecimonas xiamenensis),其几丁质酶酶活力为0.631 U/mL,最适反应温度为35 ℃,最适反应pH为6.0, Na+、Ca2+、Mn2+、K+对Chi34几丁质酶具有显著的激活作用,Cu2+、Fe3+、Ba2+、Zn2+、Cd2+、Co2+ 、EDTA和SDS等对Chi34几丁质酶有抑制作用,Chi34几丁质酶降解几丁质胶体得到的产物主要是几丁二糖和少量的几丁单糖。结论:本研究的几丁质酶能催化几丁质胶体产生几丁二糖和几丁单糖,可为几丁质的高值化利用提供一定的理论参考。

     

    Abstract: Objective: The marine ecosystem contains the most chitin in nature, which incubates a large number of microorganisms that can degrade and utilize chitin, therefore, the degradation of chitin by marine microbial chitinase is an important way to produce high value-added chitooligosaccharides. The aim of this study was to screen a stable chitinase-producing strain from the marine mud in Lianyungang. Methods: The chitinase-producing strain Chi34 was screened by plate screening and flask shaking fermentation re-screening and identified by morphological analysis and 16S rDNA sequence analysis. Then, effects of temperature, pH, metal ions, EDTA and SDS on Chi34 chitinase were studied. Finally, the TLC experiment was taken to analyze products of colloidal chitin hydrolyzed by Chi34 chitinase. Result: The isolated strain Chi34 was identified as Gallaecimonas xiamenensis and the activity of Chi34 chitinase was 0.631 U/mL. The optimum pH and temperature for the chitinolytic activity were 6.0 and 35 ℃, respectively. Chi34 chitinase was highly activated by Na+, Ca2+, Mn2+, and K+, while inhibited by Cu2+, Fe3+, Ba2+, Zn2+, Cd2+, Co2+, EDTA, and SDS. The main products of chitin hydrolyzed by Chi34 chitinase were (GlcNAc)2 with little GlcNAc. Conclusion: The Chi34 chitinase could hydrolyze chitin to produce (GlcNAc)2 and GlcNAc, which would provide a certain theoretical reference for the high value utilization of chitin.

     

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