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一种岩藻依聚糖降解海洋细菌菌株及其岩藻依聚糖酶的分离与鉴定

Isolation and characterization of a fucoidan-degrading marine bacterial strain and its fucoidanase.

作者信息

Sakai Takeshi, Kawai Takashi, Kato Ikunoshin

机构信息

Takara Bio Inc., Biotechnology Research Laboratories, Seta 3-4-1, Otsu-shi, Shiga-ken 520-2193, Japan.

出版信息

Mar Biotechnol (NY). 2004 Jul-Aug;6(4):335-46. doi: 10.1007/s10126-003-0033-5. Epub 2004 Jun 23.

Abstract

A marine bacterial strain that degraded fucoidan from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) was isolated in our laboratory. The strain was gram-negative, ubiquinone 8 was the predominant respiratory quinone, and the GC-content of its genomic DNA was 36%. The cells of the strain were rod-shaped (2.0 microm long x 1.0 microm wide), and each cell was motile by means of one polar flagellum. Phylogenetic analysis of its 16S ribosomal DNA sequence indicated that it was a member of the family Alteromonadaceae. It produced a type of extracellular fucoidanase, an endosulfated fucan-digesting enzyme. The enzyme was purified with 3500-fold purity at 12.0% yield. Optimum conditions for the enzyme reaction were approximately pH 6.5 to 8.0 and temperature 30 degrees to 35 degrees C. The enzyme was activated by calcium ions, and maximum activity was observed in the presence of greater than 30 mM calcium ion.

摘要

我们实验室分离出了一株能降解厚叶海带(褐藻纲、海带目、海带科)中岩藻依聚糖的海洋细菌菌株。该菌株为革兰氏阴性菌,主要呼吸醌为泛醌8,其基因组DNA的鸟嘌呤-胞嘧啶含量为36%。该菌株的细胞呈杆状(长2.0微米×宽1.0微米),每个细胞通过一根极生鞭毛运动。对其16S核糖体DNA序列的系统发育分析表明,它是交替单胞菌科的一员。它产生了一种胞外岩藻依聚糖酶,一种能消化硫酸化岩藻聚糖的内切酶。该酶经纯化后纯度提高了3500倍,产率为12.0%。酶反应的最佳条件约为pH 6.5至8.0,温度为30℃至35℃。该酶被钙离子激活,在钙离子浓度大于30 mM时观察到最大活性。

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