Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5

被引:4
作者
Chao, Yaxi [1 ]
Wang, Shuyan [1 ]
Wu, Suqi [1 ,2 ]
Wei, Jiaqiang [1 ]
Chen, Hao [1 ,2 ]
机构
[1] Minist Nat Resources, Inst Oceanog 1, Key Lab Marine Bioact Subst, Qingdao 266061, Shandong, Peoples R China
[2] Pilot Natl Lab Marine Sci & Technol Qingdao, Lab Marine Biol & Biotechnol, Qingdao 266237, Shandong, Peoples R China
关键词
alginate lyase; Vibrio sp; alginate; poly--L-guluronic acid lyase; oligosaccharides; OLIGOSACCHARIDE; PURIFICATION; AGARIVORANS;
D O I
10.1007/s13131-018-1226-0
中图分类号
P7 [海洋学];
学科分类号
0707 ;
摘要
Seven bacterial clones with alginate-utilizing activity were isolated from rotten kelp. By activity test, the Vibrio sp. QD-5 with the potential alginate-degrading capability was chosen to carry out the draft genome sequencing, and the result showed that the Vibrio sp. QD-5 containing an alginate lyase gene cluster. One of these genes, aly-IV, was cloned and characterized for the first time. After overexpression, Aly-IV, with a molecular mass of about 62 kDa and a theoretical isoelectric point (pI) of 5.12, was purified to a specific activity of 1 256.78 U/mg and showed highest activity at 35 degrees C in the Tris-HCl buffer at pH of 8.9. Moreover, the enzyme activity was enhanced by the metal ions of Na+, K+ and Mg2+ under certain concentration. Aly-IV degraded favorably polyG blocks in an endo-type, yielding monomer and dimer as the main products. Due to its high substrate specificity, Aly-IV could be used as a potential tool for production of polyG oligosaccharides with low degree of polymerization (DP) and for determining the fine structure of alginate.
引用
收藏
页码:65 / 74
页数:10
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