Characterization of a NaCl-tolerant β-N-acetylglucosaminidase from Sphingobacterium sp. HWLB1.
β-N-Acetylglucosaminidases serve important biological functions and various industrial applications. A glycoside hydrolase family 3 β-N-acetylglucosaminidase gene was cloned from Sphingobacterium sp. HWLB1 and expressed in Escherichia coli BL21 (DE3). The purified recombinant enzyme (rNag3HWLB1) showed apparent optimal activity at pH 7.0 and 40 °C. In the presence of 0.5-20.0 % (w/v) NaCl, the activity and stability of rNag3HWLB1 were slightly affected or not affected. The enzyme could even retain 73.6 % activity when 30.0 % (w/v) NaCl was added to the reaction mixture. The half-life of the enzyme was approximately 10 min at 37 °C without the addition of NaCl. However, the enzyme was stable at 37 °C in the presence of 3.0 % (w/v) NaCl. A large negatively charged surface in the catalytic pocket of the enzyme was observed and might contribute to NaCl tolerance and thermostability improvement. The degree of synergy between a commercial endochitinase and rNag3HWLB1 on chitin enzymatic degradation ranged from 3.11 to 3.74. This study is the first to report the molecular and biochemical properties of a NaCl-tolerant β-N-acetylglucosaminidase.
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Junpei
J
Zhou
College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, Kunming, Yunnan, 650500, People's Republic of China.
Zhifeng
Z
Song
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Rui
R
Zhang
College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, Kunming, Yunnan, 650500, People's Republic of China.
Limei
L
Ding
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Qian
Q
Wu
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Junjun
J
Li
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Xianghua
X
Tang
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Bo
B
Xu
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Junmei
J
Ding
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, 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, 650500, People's Republic of China.
Nanyu
N
Han
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China. huangzunxi@163.com.|College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, Kunming, Yunnan, 650500, People's Republic of China. huangzunxi@163.com.|Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, Yunnan, 650500, People's Republic of China. huangzunxi@163.com.|Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, People's Republic of China. huangzunxi@163.com.
Zunxi
Z
Huang
D000118:Acetylglucosaminidase / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D020134:Catalytic Domain; D002686:Chitin / Q000378:metabolism; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D055049:Salt Tolerance; D012965:Sodium Chloride; D045249:Sphingobacterium / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D002686:Chitin; D012965:Sodium Chloride; D000118:Acetylglucosaminidase
NaCl tolerance; Negatively charged surface; Sphingobacterium; Synergistic action; Thermostability; β-N-Acetylglucosaminidase