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微生物学报

褐藻胶降解菌的筛选、鉴定及产酶条件优化
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海洋公益性行业科研专项(201505026-5);福建省高校产学合作重大项目(2018N5008);泉州市科技计划重点项目(2015Z140)


Screening, identification and fermentation optimization of an alginate-degrading strain
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    摘要:

    [目的]筛选一株能降解褐藻胶的菌株,并优化产酶条件以提高褐藻胶裂解酶活力。[方法]从漳州海域采集到海水和海泥,以海藻酸钠为唯一碳源,通过富集培养、初筛、复筛筛选到一株能够降解褐藻胶的菌株。依据16S rRNA序列分析、生理生化特征、菌体形态及菌落特征对该菌进行鉴定。通过单因素和正交试验对该菌的产酶条件进行优化。[结果]该菌属于海科贝特氏菌,命名为Cobetia marina HQZ08。该菌株最佳的产酶培养基组成为:海藻酸钠7.00 g/L、蛋白胨3.00 g/L、NaCl 30.00 g/L,K2HPO4·3H2O 1.25 g/L。最佳发酵条件为:接种量2%,接种龄12 h,培养基起始pH为7.0,培养温度25℃,培养时间24 h。优化后褐藻胶裂解酶活力达到68.5 U/mL,TLC法分析酶解产物为褐藻胶寡糖。[结论]HQZ08菌株可以用于降解褐藻胶,产生聚合度为2-6的褐藻胶寡糖。

    Abstract:

    [Objective] To screen an alginate-degrading strain and to improve the activity of its alginate lyase by optimizing the fermentation conditions.[Methods] A strain capable of degrading alginate was screened by the steps of enrichment culture, screening, and re-screening by using seawater and sea mud collected from the sea area of Zhangzhou (Fujian, China). Then, the strain was identified according to its 16S rRNA sequence analysis, physiological and biochemical characteristics, mycelium morphology and colony characteristics. The conditions of enzyme production of the strain were optimized by single factor and orthogonal test. [Results] The strain, belonging to the genus Cobetia marina, was named Cobetia marina HQZ08. The optimal medium of the strain was composed of 7.00 g/L sodium alginate, 3.00 g/L peptone, 30.00 g/L NaCl, 1.25 g/L K2HPO4·3H2O. The optimal fermentation conditions were as follows:the optimum inoculum was 2%, the inoculation age was 12 h, the initial pH of the culture medium was 7.0, the culture temperature was 25℃, and the incubation time was 24 h. The enzymatic activity of alginate lyase reached 68.5 U/mL after optimization. Enzymatic hydrolysates were determined to be alginate oligosaccharides. [Conclusion] HQZ08 strain can be used to degrade alginate to produce alginate oligosaccharides with a degree of polymerization of 2-6.

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赵婉琳,叶静,张娜,肖美添,赵鹏,黄雅燕. 褐藻胶降解菌的筛选、鉴定及产酶条件优化. 微生物学报, 2019, 59(1): 169-180

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  • 收稿日期:2018-03-10
  • 最后修改日期:2018-07-05
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  • 在线发布日期: 2018-12-29
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