title: Characterization of a Glycoside Hydrolase Family 27 α-Galactosidase from Pontibacter Reveals Its Novel Salt-Protease Tolerance and Transglycosylation Activity.
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.
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Junpei
authors.initials: J
authors.lastname: Zhou
authors.affiliation: College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Lu
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wu
authors.affiliation: College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Li
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Li
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Xianghua
authors.initials: X
authors.lastname: Tang
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Junmei
authors.initials: J
authors.lastname: Ding
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Nanyu
authors.initials: N
authors.lastname: Han
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Kunming, Yunnan 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University , Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Zunxi
authors.initials: Z
authors.lastname: Huang
mesh_terms: D041963:Bacteroidetes / Q000201:enzymology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006031:Glycosylation; D000439:Pentanols / Q000378:metabolism; D011887:Raffinose / Q000378:metabolism; D011994:Recombinant Proteins; D012965:Sodium Chloride / Q000494:pharmacology*; D013395:Sucrose / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology*; D000519:alpha-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D000439:Pentanols; D011994:Recombinant Proteins; D012965:Sodium Chloride; D013395:Sucrose; D000519:alpha-Galactosidase; D014357:Trypsin; D011887:Raffinose
keywords: glycoside hydrolase family 27; protease tolerance; salt tolerance; transglycosylation; α-galactosidase