Biochemical Characterization and Degradation Pattern of a Novel Endo-Type Bifunctional Alginate Lyase AlyA from Marine Bacterium Isoptericola halotolerans.
Biochemical Characterization and Degradation Pattern of a Novel Endo-Type Bifunctional Alginate Lyase AlyA from Marine Bacterium Isoptericola halotolerans.
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海洋细菌等孢菌的新型内型双功能藻酸盐裂解酶 AlyA 的生化特征和降解模式
DOI：
10.3390/md16080258
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发表时间：
2018-07-31
期刊：
影响因子：
5.4
通讯作者：
Ge L
中科院分区：
文献类型：
--
作者：
Zhu B;Ning L;Jiang Y;Ge L
褐藻胶裂解酶是降解褐藻胶制备具有多种生理活性寡糖的重要工具。特别地，双功能海藻酸裂解酶可以有效地将多糖水解成寡糖。本文从耐盐等翅菌NJ-05中克隆并鉴定了一种新的双功能藻酸盐裂解酶AlyA，该酶具有高活性和广泛的底物特异性，可用于制备低聚糖。为了进一步在工业上应用，该酶已被表征，其作用模式也已阐明。在pH7.5和55 °C时，酶活最高（7984.82 U/mg）。此外，它具有广泛的底物特异性，不仅对polyM（聚β-d-甘露糖醛酸）（7658.63 U/mg）显示出高活性，而且对polyG（聚α-l-古洛糖醛酸）（8643.29 U/mg）也显示出高活性。此外，AlyA对polyG（3.2mM）的Km值低于对海藻酸钠（5.6mM）和polyM（6.7mM）的Km值。利用薄层色谱（TLC）和电喷雾电离质谱（ESI-MS）研究了该酶的作用方式，结果表明该酶以内溶方式水解底物，释放出二糖、三糖和四糖等一系列寡糖。该研究为深入了解海藻酸裂解酶的底物识别和降解模式提供了新的思路，具有广泛的底物特异性。
Alginate lyases are important tools to prepare oligosaccharides with various physiological activities by degrading alginate. Particularly, the bifunctional alginate lyase can efficiently hydrolyze the polysaccharide into oligosaccharides. Herein, we cloned and identified a novel bifunctional alginate lyase, AlyA, with a high activity and broad substrate specificity from bacterium Isoptericola halotolerans NJ-05 for oligosaccharides preparation. For further applications in industry, the enzyme has been characterized and its action mode has been also elucidated. It exhibited the highest activity (7984.82 U/mg) at pH 7.5 and 55 °C. Additionally, it possessed a broad substrate specificity, showing high activities towards not only polyM (polyβ-d-mannuronate) (7658.63 U/mg), but also polyG (poly α-l-guluronate) (8643.29 U/mg). Furthermore, the Km value of AlyA towards polyG (3.2 mM) was lower than that towards sodium alginate (5.6 mM) and polyM (6.7 mM). TLC (Thin Layer Chromatography) and ESI-MS (Electrospray Ionization Mass Spectrometry) were used to study the action mode of the enzyme, showing that it can hydrolyze the substrates in an endolytic manner to release a series of oligosaccharides such as disaccharide, trisaccharide, and tetrasaccharide. This study provided extended insights into the substrate recognition and degrading pattern of the alginate lyases, with a broad substrate specificity.
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影响因子：
2.7
作者：
Kim, Hee Taek;Ko, Hyeok-Jin;Kim, Kyoung Heon
通讯作者：
Kim, Kyoung Heon
影响因子：
3.9
作者：
OTTERLEI, M;OSTGAARD, K;ESPEVIK, T
通讯作者：
ESPEVIK, T
DOI：
10.1016/0005-2744(74)90464-1
发表时间：
1974-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
ELYAKOVA, LA;FAVOROV, VV
通讯作者：
FAVOROV, VV
影响因子：
5
作者：
Haraguchi, K;Kodama, T
通讯作者：
Kodama, T
影响因子：
3.3
作者：
Inoue, Akira;Kagaya, Mayu;Ojima, Takao
通讯作者：
Ojima, Takao