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Efficient Over-expression and Application of High-performance Pectin Lyase by Screening Aspergillus niger Pectin Lyase Gene Family

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Abstract

Acidic pectin lyase is of great application value in fruit-processing industry. In this study, five pectin lyase genes (pelA, pelB, pelC, pelD and pelF) from A. niger pectin lyase gene family were cloned and over-expressed in A. niger strain SH-2, respectively. The pelA recombinant strain (SH2-pelA) owns the highest acidic pectin lyase activity: 11069.2 U/mL in flask fermentation, and 65148.8 U/mL in submerged fermentation. The recombinant pectin lyase A (PelA) was purified by Ni-affinity chromatography, and specificity of the protein was confirmed by western blotting. The recombinant PelA exhibits maximum activity at pH 4.5 and 50°C. The recombineant PelA shows increasing specific activity as the degree of methyl-esterization of the substrate raises. The maximum activity (22423.3 U/mL) was obtined using citrus pectin (≥85% esterified) as the substrate. After the treatment by PelA, the light transmittance of orange, apple and grape juice was increased 19.2-fold, 7.3-fold and 3.8-fold, respectively, suggesting that the recombinant acidic PelA had noticeable clarifying effects on orange, apple and grape juice.

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Authors and Affiliations

  1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, China

    Yulan He, Li Pan & Bin Wang

  2. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, 510006, China

    Li Pan & Bin Wang

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  1. Yulan He
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  2. Li Pan
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  3. Bin Wang
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Correspondence to Bin Wang.

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He, Y., Pan, L. & Wang, B. Efficient Over-expression and Application of High-performance Pectin Lyase by Screening Aspergillus niger Pectin Lyase Gene Family. Biotechnol Bioproc E 23, 662–669 (2018). https://doi.org/10.1007/s12257-018-0387-1

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