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中国精品科技期刊2020
梁颖,刘睿,宋超东,等. 高产壳聚糖酶菌株筛选鉴定、条件优化及酶学性质分析[J]. 食品工业科技,2024,45(20):157−166. doi: 10.13386/j.issn1002-0306.2023110321.
引用本文: 梁颖,刘睿,宋超东,等. 高产壳聚糖酶菌株筛选鉴定、条件优化及酶学性质分析[J]. 食品工业科技,2024,45(20):157−166. doi: 10.13386/j.issn1002-0306.2023110321.
LIANG Ying, LIU Rui, SONG Chaodong, et al. Screening, Identification and Conditions Optimization of High Chitosanase Producing Strains, and Analysis of Its Enzymatic Properties[J]. Science and Technology of Food Industry, 2024, 45(20): 157−166. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110321.
Citation: LIANG Ying, LIU Rui, SONG Chaodong, et al. Screening, Identification and Conditions Optimization of High Chitosanase Producing Strains, and Analysis of Its Enzymatic Properties[J]. Science and Technology of Food Industry, 2024, 45(20): 157−166. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110321.

高产壳聚糖酶菌株筛选鉴定、条件优化及酶学性质分析

Screening, Identification and Conditions Optimization of High Chitosanase Producing Strains, and Analysis of Its Enzymatic Properties

  • 摘要: 为了筛选出高产壳聚糖酶的菌株,并对其最优产酶条件及酶学性质进行研究。本研究从广西北海养殖虾塘底泥采样,经初筛、复筛选出高产壳聚糖酶菌株,并进行鉴定,再通过单因素实验、Plackett-Burman 及正交试验优化产酶工艺,最后对其酶学性质及产物进行研究。结果表明,筛选到1株高产壳聚糖酶的菌株Gxun-5.3.3,鉴定为蜡样芽孢杆菌(Bacillus cereus);菌株Gxun-5.3.3产酶的最优工艺为:麦芽糖5 g/L、酵母粉3 g/L、壳聚糖 15 g/L、初始pH6.0、发酵时间84 h,优化后酶活可达62.03±1.18 U/mL,较优化前(39.87±1.24 U/mL)提高了55.58%;酶学性质研究表明,该酶最适反应温度为60 ℃,最适反应pH为5.0,金属离子中Fe2+、Mn2+对壳聚糖酶有显著(P<0.05)的激活作用,而Al3+、Cu2+则有较强的抑制作用;酶解壳聚糖产物经薄层层析(Thin Layer Chromatography,TLC)测定为聚合度2~4的壳寡糖,表明菌株所产壳寡糖酶为内切型。本研究为B. cereus Gxun-5.3.3壳聚糖酶的生产和壳寡糖的酶法制备提供了理论依据。

     

    Abstract: To screen and identify high chitosanase producing strains and to optimize the fermentation condition and analyze the enzymatic properties of the target strain. In this study, high chitosanase-producing strain was screened from the sediment of shrimp ponds in Beihai, Guangxi through primary and secondary screening. The strain underwent identification, the enzyme production process of the strain was optimized through single-factor, Plackett-Burman and orthogonal optimization methods. The enzymatic properties and products of chitosanase produced by the strain were studied. The results showed that the strain of Gxun-5.3.3 with high chitosanase production was obtained by screening and was identified as Bacillus cereus. The optimal components for the enzyme-producing medium of B. cereus Gxun-5.3.3 was maltose 5 g/L, yeast powder 3 g/L, chitosan 15 g/L, and the initial pH and fermentation time were 6.0 and 84 h, respectively. Under the optimized conditions, the chitosanase activity reached 62.03±1.18 U/mL, which was 55.58% higher before optimization (39.87±1.24 U/mL). The protease produced by strain Gxun-5.3.3 had maximum activity at 60 ℃ and optimum pH at 5.0, respectively. Metal ions Fe2+ and Mn2+ had a significant (P<0.05) activation effect on chitosanase, but Al3+ and Cu2+ had a strong inhibition. The enzymatic chitosan products were determined by TLC (thin layer chromatography) as chito-oligosaccharides with polymerization degree 2~4, indicating that the chitosanase produced by the strain was acting on endo-oligosaccharides. This study would provide a theoretical basis for the chitosanase production of B. cereus Gxun-5.3.3 and the enzymatic preparation of chito-oligosaccharides.

     

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