[Construction of Aspergillus niger lipase mutants with oil-water interface independence].
Based on previous bioinformational analysis results, two Aspergillus niger lipase (ANL) mutants, ANL-Ser84Gly and ANL-Asp99Pro were constructed to screen ANL mutants with oil-water interface independence. ANL-Ser84Gly still displayed a pronounced interfacial activation, while ANL-Asp99Pro displayed no interfacial activation. The specific activity of ANL-Ser84Gly towards p-nitrophenyl palmitate (-myristate, -laurate and -decanoate) decreased by 29.8% (53.1, 60.1 and 77.1, respectively) than that of ANL, while the specific activity of ANL-Asp99Pro towards p-nitrophenyl palmitate increased by 2.2-fold. The mutation in the hinge region at both sides of the lid domain also destabilized various secondary structure factors of ANL-S84G and ANL-D99P, which resulted in a substantial decrease in thermostability. The achievement to construct oil-water interface-independent ANL mutants would help to further understand lipase interfacial activation mechanism.
Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China.
De
D
Chen
Zhengyu
Z
Shu
Longyin
L
Xue
Ruifeng
R
Lin
Jiguang
J
Wu
Yongmei
Y
Jiang
Xin
X
Li
Yuexin
Y
Lin
Jianzhong
J
Huang
D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000235:genetics*; D009821:Oils; D013379:Substrate Specificity; D014867:Water
D005656:Fungal Proteins; D050505:Mutant Proteins; D009821:Oils; D014867:Water; D008049:Lipase