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KMID : 0545120200300020196
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Journal of Microbiology and Biotechnology
2020 Volume.30 No. 2 p.196 ~ p.205
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Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
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Zhu Si-Yuan
Xu Yan
Yu Xiao-Wei
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Abstract
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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.
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KEYWORD
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Aspergillus niger, acidic lipase, expression, characterization, Agrobacterium-mediated transformation
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