A novel surfactant-, NaCl-, and protease-tolerant β-mannanase from Bacillus sp. HJ14.
A glycoside hydrolase family 5 β-mannanase-encoding gene was cloned from Bacillus sp. HJ14 isolated from saline soil in Heijing town. Coding sequence of mature protein (without the predicted signal peptide from M1 to A30) was successfully expressed in Escherichia coli BL21 (DE3). Purified recombinant mannanase (rMan5HJ14) exhibited optimal activity at pH 6.5 and 65 °C. The enzyme showed good salt tolerance, retaining more than 56 % β-mannanase activity at 3.0-30.0 % (w/v) NaCl and more than 94 % of the initial activity after incubation with 3.0-30.0 % (w/v) NaCl at 37 °C for 60 min. Almost no mannanase activity was lost after incubation of rMan5HJ14 with trypsin, proteinase K, and Alcalase at 37 °C for 60 min. Surfactants and chelating agents, namely SDS, CTAB, Tween 80, Triton X-100, EDTA, and sodium tripolyphosphate, showed little or no effect (retaining >82.4 % activity) on enzymatic activity. Liquid detergents, namely Tupperware, Walch, Bluemoon, Tide, and OMO, also showed little or no effect (retaining >72.4 % activity) on enzymatic activity at 0.5-2.0 % (v/v). The enzyme further presents a high proportion (11.97 %) of acidic amino acid residues (D and E), which may affect the SDS and NaCl tolerance of the enzyme. Together, the mannanase may be an alternative for potential use in liquid detergent industry.
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, Yunnan, Kunming, 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.
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, Yunnan, Kunming, 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, Yunnan, Kunming, 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, People's Republic of China.
Junpei
J
Zhou
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, Yunnan, Kunming, 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, Yunnan, Kunming, 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, People's Republic of China.
Yuelin
Y
Mu
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, Yunnan, Kunming, 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, Yunnan, Kunming, 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, Yunnan, Kunming, 650500, People's Republic of China.|Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, People's Republic of China.
Junmei
J
Ding
College of Life Sciences, Yunnan Normal University, No.1 Yuhua District, Chenggong, Kunming, Yunnan, 650500, People's Republic of China.
Shucan
S
Deng
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, Yunnan, Kunming, 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
D000595:Amino Acid Sequence; D001407:Bacillus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D015870:Gene Expression; D016680:Genome, Bacterial; D006868:Hydrolysis; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D055049:Salt Tolerance; D017422:Sequence Analysis, DNA; D012965:Sodium Chloride / Q000494:pharmacology*; D013501:Surface-Active Agents / Q000494:pharmacology*; D044902:beta-Mannosidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D003902:Detergents; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D012965:Sodium Chloride; D044902:beta-Mannosidase; D010450:Endopeptidases