1. Characterization of a Glycoside Hydrolase Family 27 α-Galactosidase from Pontibacter Reveals Its Novel Salt–Protease Tolerance and Transglycosylation Activity
- Author
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Chunyan Li, Xu Bo, Ding Junmei, Junpei Zhou, Junjun Li, Xianghua Tang, Yu Liu, Rui Zhang, Qian Lu, Qian Wu, Zunxi Huang, and Han Nanyu
- Subjects
0301 basic medicine ,Sucrose ,Glycosylation ,medicine.medical_treatment ,Gene Expression ,Sodium Chloride ,03 medical and health sciences ,chemistry.chemical_compound ,Pentanols ,Raffinose ,Escherichia coli ,medicine ,Trypsin ,Glycoside hydrolase ,Cloning, Molecular ,chemistry.chemical_classification ,Guar gum ,Protease ,Alpha-galactosidase ,biology ,Bacteroidetes ,General Chemistry ,Recombinant Proteins ,030104 developmental biology ,Enzyme ,chemistry ,Biochemistry ,alpha-Galactosidase ,biology.protein ,Locust bean gum ,General Agricultural and Biological Sciences ,medicine.drug - Abstract
α-Galactosidases are of great interest in various applications. A glycoside hydrolase family 27 α-galactosidase was cloned from Pontibacter sp. harbored in a saline soil and expressed in Escherichia coli. The purified recombinant enzyme (rAgaAHJ8) was little or not affected by 3.5-30.0% (w/v) NaCl, 10.0-100.0 mM Pb(CH3COO)2, 10.0-60.0 mM ZnSO4, or 8.3-100.0 mg mL(-1) trypsin and by most metal ions and chemical reagents at 1.0 and 10.0 mM concentrations. The degree of synergy on enzymatic degradation of locust bean gum and guar gum by an endomannanase and rAgaAHJ8 was 1.22-1.54. In the presence of trypsin, the amount of reducing sugars released from soybean milk treated by rAgaAHJ8 was approximately 3.8-fold compared with that treated by a commercial α-galactosidase. rAgaAHJ8 showed transglycosylation activity when using sucrose, raffinose, and 3-methyl-1-butanol as the acceptors. Furthermore, potential factors for salt adaptation of the enzyme were presumed.
- Published
- 2016