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Genome sequencing of Aureobasidium pullulans P25 and overexpression of a glucose oxidase gene for hyper-production of Ca2+-gluconic acid

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Abstract

Gluconic acid (GA) has many applications such as in the food and pharmaceutical industry. Aureobasidium pullulans P25 strain is able to produce high levels of Ca2+-GA. The genome length, GC content and the gene number of this yeast were found to be 30.97 Mb, 50.28% and 10,922, respectively. The pathways for gluconic acid biosynthesis were annotated. Glucose oxidase (Gox) sequences from different strains of A. pullulans were highly similar but were distinct from those of other fungi. The glucose oxidase had two FAD binding sites and a signal sequence. Deletion of the GOX gene resulted in a strain that showed no Gox activity and that was unable to produce Ca2+-GA. Overexpression of the GOX gene in strain P25 generated strain GA-6 that produced 200.2 ± 2.3 Ca2+-GA g/l and 2480 U/mg of Gox activity. The productivity of Ca2+-GA was 2.78 g/l/h and the yield was 1.1 g/g.

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Acknowledgements

The authors gratefully acknowledge the support rendered by the National Natural Science Foundation of China (Grant No. 31770061).

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Authors and Affiliations

  1. College of Marine Life Sciences, Ocean University of China, Yushan Road, No. 5, Qingdao, China

    Shou-Feng Zhao, Hong Jiang, Zhe Chi, Guang-Lei Liu, Zhen-Ming Chi, Tie-Jun Chen & Guang Yang

  2. Department of Biology, Shantou University, Shantou, 515063, China

    Zhong Hu

Authors
  1. Shou-Feng Zhao
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  2. Hong Jiang
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  3. Zhe Chi
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  4. Guang-Lei Liu
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  5. Zhen-Ming Chi
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  6. Tie-Jun Chen
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  7. Guang Yang
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  8. Zhong Hu
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Contributions

S-FZ and HJ carried out all the experiments, TJC and GY were responsible for drawing some figures, ZC and G-LL were involved in DNA sequence and annotation, Z-MC and ZH wrote the manuscript.

Corresponding authors

Correspondence to Zhe Chi or Zhen-Ming Chi.

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The authors declare that they have no conflict of interests.

Electronic supplementary material

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Supplementary file 1

The primers used in this study. (DOC 40 kb)

Supplementary file 2

Construction of the vectors for disruption (A) and expression (B) of the GOX gene in A. pullulans P25. (DOC 107 kb)

Supplementary file 3

The nucleotide sequence of the P25 glucose oxidase gene and its bordering regions and amino acid sequence deduced from the glucose oxidase gene. Translation initiation codon ATG was positioned at nt +1. The gene encoded 606 amino acids. The promoter region containing TATA box was underlined. 5′-SYGGRG-3′ motif was shaded and N-glycosylation sites were shaded. FAD binding sites were underlined and signal sequence was shaded and marked. (DOC 579 kb)

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Zhao, SF., Jiang, H., Chi, Z. et al. Genome sequencing of Aureobasidium pullulans P25 and overexpression of a glucose oxidase gene for hyper-production of Ca2+-gluconic acid. Antonie van Leeuwenhoek 112, 669–678 (2019). https://doi.org/10.1007/s10482-018-1197-3

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  • DOI: https://doi.org/10.1007/s10482-018-1197-3

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