Molecular cloning and expression of a new -neoagarobiose hydrolase from Agarivorans gilvus WH0801 and enzymatic production of 3,6-anhydro-l-galactose
Molecular cloning and expression of a new -neoagarobiose hydrolase from Agarivorans gilvus WH0801 and enzymatic production of 3,6-anhydro-l-galactose
复制标题
来自 Agarivorans gilvus WH0801 的新型 α-新琼脂二糖水解酶的分子克隆和表达以及 3,6-脱水-L-半乳糖的酶促生产
DOI：
10.1002/bab.1363
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发表时间：
2016-03-01
影响因子：
2.8
通讯作者：
Wei, Dongzhi
中科院分区：
文献类型：
--
作者：
Liu, Nan;Yang, Meng;Wei, Dongzhi
A new -neoagarobiose hydrolase (NABH) called AgaWH117 was cloned from Agarivorans gilvus WH0801. The gene encoding this hydrolase consists of 1,086bp and encodes a protein containing 361 amino acids. This new NABH showed 74% amino acid sequence identity with other known NABHs. The molecular mass of the recombinant AgaWH117 was estimated to be 41kDa. Purified AgaWH117 showed endolytic activity during neoagarobiose degradation, yielding 3,6-anhydro-l-galactose (l-AHG) and d-galactose as products. It showed a maximum activity at a temperature of 30 degrees C and a pH of 6.0 and was stable at temperatures below 30 degrees C. Its K-m and V-max values were 2.094mg/mL and 6.982U/mg, respectively. The cloning strategy used and AgaWH117 isolated in this study will provide information on the saccharification process of marine biomass. This study provides a method to produce l-AHG from agarose by using AgaWH117 without an acid and describes its one-step purification by using Bio-Gel P2 chromatography.
