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Expression, purification and characterization of pectate lyase A from Aspergillus nidulans in Escherichia coli

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Abstract

Pectate lyase A (PelA) of Aspergillus nidulans was successfully expressed in Escherichia coli and effectively purified using a Ni2+-nitrilotriacetate-agarose column. Enzyme activity of the recombinant PelA could reach 360 U ml−1 medium. The expressed PelA exhibited its optimum level of activity over the range of pH 7.5–10 at 50°C. Mn2+, Ca2+, Fe2+, Mg2+ and Fe3+ ions stimulated the pectate lyase activity, but Cu2+ and Zn2+ inhibited it. The recombinant PelA had a V max of 77 μmol min−1 mg−1 and an apparent K m of 0.50 mg ml−1 for polygalacturonic acid. Low-esterified pectin was the optimum substrate for the PelA, whereas higher-esterified pectin was hardly cleaved by it. PelA efficiently macerated mung bean hypocotyls and potato tuber tissues into single cells.

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Acknowledgments

This work was supported by the funds from the National Natural Science Foundation of China (30170005) and the Natural Science Foundation of Jiangsu Province (BK2005136).

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Authors and Affiliations

  1. Jiangsu Key Laboratory for Biodiversity and Biotechnology, Key Lab for Microbial Technology in the College of Life Science, Nanjing Normal University, Nanjing, 210097, People’s Republic of China

    Qingxin Zhao, Sheng Yuan, Yuling Zhang, Hong Zhu, Chuanchao Dai, Fang Yang & Fengmin Han

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  1. Qingxin Zhao
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  2. Sheng Yuan
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  3. Yuling Zhang
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  4. Hong Zhu
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  5. Chuanchao Dai
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  6. Fang Yang
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  7. Fengmin Han
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Correspondence to Sheng Yuan.

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Zhao, Q., Yuan, S., Zhang, Y. et al. Expression, purification and characterization of pectate lyase A from Aspergillus nidulans in Escherichia coli . World J Microbiol Biotechnol 23, 1057–1064 (2007). https://doi.org/10.1007/s11274-006-9331-1

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  • DOI: https://doi.org/10.1007/s11274-006-9331-1

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