2019 ; Vol.29-2: 235~243
|Author||Yan Liu, Xiaoxu Tian, Chao Peng, Zongjun Du|
|Place of duty||College of Marine Science, Shandong University, P.R. China|
|Title||Isolation and Characterization of an Eosinophilic GH 16 β-Agarase (AgaDL6) from an Agar-Degrading Marine Bacterium Flammeovirga sp. HQM9|
J. Microbiol. Biotechnol.2019 ;
|Abstract||A special eosinophilic agarase exo-type β-agarase gene, AgaDL6, was cloned from a marine
agar-degrading bacterium, Flammeovirga sp. HQM9. The gene comprised 1,383-bp nucleotides
encoding a putative agarase AgaDL6 of 461 amino acids with a calculated molecular mass of
52.8 kDa. Sequence analysis revealed a β-agarase domain that belongs to the glycoside
hydrolase family (GH) 16 and a carbohydrate-binding module (CBM_4_9) unique to agarases.
AgaDL6 was heterologously expressed in Escherichia coli BL21 (DE3). Enzyme activity analysis
of the purified protein showed that the optimal temperature and pH of AgaDL6 were 50°C
and 3.0, respectively. AgaDL6 showed thermal stability by retaining more than 98% of activity
after incubation for 2 h at 50°C, a feature quite different from other agarases. AgaDL6 also
exhibited outstanding acid stability, retaining 100% of activity after incubation for 24 h at pH
2.0 to 5.0, a property distinct from other agarases. This is the first agarase characterized to
have such high acid stability. In addition, we observed no obvious stimulation or inhibition of
AgaDL6 in the presence of various metal ions and denaturants. AgaDL6 is an exo-type β-1,4
agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products.
These characteristics make AgaDL6 a potentially valuable enzyme in the cosmetic, food, and
|Key_word||Agarase, GH16, CBM_4_9, eosinophilic, thermostable enzyme|
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