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Optimization of the production of Aspergillus niger α-glucosidase expressed in Pichia pastoris

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Abstract

The α-glucosidase (AGL) from Aspergillus niger has been applied to produce isomaltooligosaccharides. In the present study, various factors which affect the yield of recombinant AGL, produced by engineered Pichia pastoris, were investigated. The expression level reached 5.5 U ml−1 in bioreactor after optimization of parameters of initial induction cell density, induction temperature and methanol concentration. In addition, it was found that coexpression of protein disulfide isomerase (PDI) inhibited the growth of the engineered P. pastoris strains and had an adverse effect on the production of AGL, while codon optimization of native A. niger α-glucosidase encoding gene (aglu) resulted in a significant enhancement of enzyme production, which reached 10.1 U ml−1. We believe that yield of AGL is increased by codon optimization as a result of enhanced translation efficiency as well as more stable mRNA secondary structure. In contrast, PDI coexpression under the control of alcohol oxidase promoter (PAOX1) seems to be less efficient in helping disulfide bond formation in AGL while probably induce unfolded protein response, which further leads to cell apoptosis and increased protein degradation.

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Acknowledgments

This work was financially supported by the Fundamental Research Funds for the Central Universities (JUSRP211A05), the National Natural Science Foundation of China (30970057 and 31100048), the open program for key laboratory of industrial biotechnology ministry of education (KLIB-KF200904) and the 111 Project (No. 111-2-06).

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Authors and Affiliations

  1. School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Xu Liu, Dan Wu, Jing Wu & Jian Chen

  2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Xu Liu, Dan Wu, Jing Wu & Jian Chen

Authors
  1. Xu Liu
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  2. Dan Wu
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  3. Jing Wu
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  4. Jian Chen
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Correspondence to Jing Wu.

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Xu Liu and Dan Wu contributed equally to this study and share first authorship.

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Liu, X., Wu, D., Wu, J. et al. Optimization of the production of Aspergillus niger α-glucosidase expressed in Pichia pastoris . World J Microbiol Biotechnol 29, 533–540 (2013). https://doi.org/10.1007/s11274-012-1207-y

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