2020 ; Vol.30-2: 196~205
|Author||Zhu Si-Yuan, Xu Yan, Yu Xiao-Wei|
|Place of duty||Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P.R. China|
|Title||Improved Homologous Expression of the Acidic Lipase from Aspergillus niger|
J. Microbiol. Biotechnol.2020 ;
|Abstract||In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A.
niger. The expression of ANL was significantly improved by the expression of the native ANL
with the introns, the addition of the Kozak sequence and the optimization of the signal
sequences. When the cDNA sequence of ANL fused with the glaA signal was expressed under
the gpdA promoter in A. niger, no lipase activity could be detected. We then tried to improve
the expression by using the full-length ANL gene containing three introns, and the lipase
activity in the supernatant reached 75.80 U/ml, probably as a result of a more stable mRNA
structure. The expression was further improved to 100.60 U/ml by introducing a Kozak
sequence around the start codon due to a higher translation efficiency. Finally, the effects of
three signal sequences including the cbhI signal, the ANL signal and the glaA signal on the
lipase expression were evaluated. The transformant with the cbhI signal showed the highest
lipase activity (314.67 U/ml), which was 1.90-fold and 3.13-fold higher than those with the
ANL signal and the glaA signal, respectively. The acidic lipase was characterized and its
highest activity was detected at pH 3.0 and a temperature of 45ºC. These results provided
promising strategies for the production of the acidic lipase from A. niger.|
|Key_word||Aspergillus niger, acidic lipase, expression, characterization, Agrobacterium-mediated transformation|
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