| 产几丁质酶细菌的筛选、鉴定及酶学性质研究 |
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| 引用本文： | 舒玉凤,李婵娟,张丽.产几丁质酶细菌的筛选、鉴定及酶学性质研究[J].湖北农业科学,2016(15):3887-3890. |
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| 作者姓名： | 舒玉凤 李婵娟 张丽 |
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| 作者单位： | 武汉设计工程学院,武汉,430205 |
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| 摘 要： | 采用胶体几丁质平板产生透明圈法,从竹林附近的土壤中分离得到一株高产几丁质酶的优良菌株,以胶体几丁质为反应底物,对其所产的几丁质酶粗酶性质进行了初步研究。结果表明,该酶最适p H 7.5,在p H 6.5~9.5的缓冲液中放置12 h后,残余酶活仍在60%以上,在p H 7.0~8.5的缓冲液中残余酶活仍在80%以上,说明该酶在中性和碱性条件下具有较好的稳定性;该酶在30℃时酶活最高,在低于60℃的处理条件下残余酶活仍在80%以上,且80℃处理后仍有37%的残余酶活,说明该酶的热稳定性较好;配制蟹壳培养基发酵该菌,发现该菌能直接降解蟹壳产生壳寡糖,且每克蟹壳降解后可产生5.3 mg还原糖。通过16S r DNA基因序列比对,鉴定该菌为芽孢杆菌属(Bacillus sp.)。
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| 关 键 词： | 几丁质酶 酶学性质 菌种鉴定 蟹壳降解 |
Isolation and Identification of a Chitinase-producing Bacteria and Characterization of Its Chitinase |
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| Abstract: | A bacterium producing high chitinase was isolated from soil around bamboo forest by method of colloidal chitin plate. The characteristics of the crude chitinase that produced by this strain was studied by using colloidal chitin as the sub-strate. The results showed that the optimum pH of the enzyme was 7.5. The crude enzyme still had 60℅ of the highest activ-ity after overnight incubation at pH 6.5~9.5, especially it still had 80℅ of the highest activity in the buffer of pH 7.0~8.5, which indicated that the enzyme was very stable under neutral or alkaline conditions. The crude chitinase displayed the maxi-mum activity at 30℃. After incubation below 60℃ for 30 min,it remained more than 80℅ of the highest activity. And after incubation at 80℃ for 30 min,it still had 37℅ of the highest activity which indicated the chitinase also had better thermal stability. After cultivation at crab shell medium,the bacteria could directly degradate crab shell to chitosan oligosaccharide, producing 5.3 mg reducing sugar per gram of crab shells. The strain was identified as Bacillus sp. by measuring its 16S rD-NA sequence. |
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| Keywords: | chitinase enzymatic properties strain identification crab shells degradation |
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