双氧水对米赫根毛霉脂肪酶在米曲霉中重组表达的调控研究.pdfVIP

现代食品科技 Modern Food Science and Technology 2015, Vol.31, No.4 Regulation of the Recombinant Expression of a Rhizomucor miehei Lipase Gene in Aspergillus oryzae by Hydrogen Peroxide WU Xin-liang, GUO Yong (School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China) Abstract: Rhizomucor miehei lipase (RML) is an important microbial lipase with a wide range of industrial applications. The effect of hydrogen peroxide (H O ) on RML expression in A. oryzae transformant ONL1 was explored in this study. The results of quantitative 2 2 fluorescence polymerase chain reaction (QF-PCR) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS) indicated that the RML activity of the cultivated A. oryzae ONL1 transformants was increased by five-fold after treatment with 10 mM H O for two hours. 2 2 The H O treatment did not appear to affect the translation of RML mRNA, with the elevated expression of RML being attributed to the H O 2 2 2 2 regulation of RML transcription. Therefore, hydrogen peroxide might regulate melO promoter-controlled heterologous gene expression at the transcriptional level. Because of the highly volatile nature of H O , a prolonged cultivation time led to a lower effect on RML activity. High 2 2 RML activity within a short cultivation time could be achieved by the continuous addition of H O in the to maintain the concentration of 10