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Effect of culture conditions on the production of ligninolytic enzymes by white rot fungi Phanerochaete chrysosporium (ATCC 20696) and separation of its lignin peroxidase

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Abstract

The present work was carried out to determine the optimum culture conditions of Phanerochaete chrysosporium (ATCC 20696) for maximizing ligninolytic enzyme production. Additionally, separation of its lignin peroxidase was conducted. After experiments, an optimized culture medium/condition was constructed (per liter of Kirk’s medium): dextrose 10 g, ammonium tartrate 0.11 g, Tween-80 0.5 g, MnSO4 7 mg, and veratryl alcohol 0.3 g in 10 mM acetic acid buffer pH 4.5. Under the optimized experimental condition, both lignin peroxidase (LiP) and manganese peroxidase (MnP) were detected and reach the highest yield at 30°C on the 8th day culture. Salt precipitation methods was used in the extraction and purification processes. Results show that salt precipitation with 60% (NH4)2SO4 yielded the best result, especially toward LiP. Enzyme separation was conducted and two fractions with LiP activity. LiP1 and LiP2 were produced using three columns sequentially: desalting column, Q FF ion exchange column and Sepharyl S-300 HR gel filtration. LiP1 and LiP2 had been purified by 9.6- and 7.6-fold with a yield of 22.9% and 18.6%, respectively. According to the data of sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE), the molecular weights of the enzymes are 38 kDa and 40 kDa, respectively.

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Acknowledgments

The research is supported by the Department of Energy (DOE-MAT-SBI Program), grant number DE-FG36-05G085002 with subcontract# 07-08-001 to JSU from University of Mississippi (P.I. Dr. Clint Williford), via management of Mr. Sumesh Arora of Mississippi Technology Alliance.

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Authors and Affiliations

  1. Biology Department, Jackson State University, Jackson, MS, 39217, USA

    Peng Wang, Xiaoke Hu, Sean Cook, Maria Begonia & Huey-Min Hwang

  2. Chemistry Department, Jackson State University, Jackson, MS, 39217, USA

    Ken S. Lee

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  1. Peng Wang
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  2. Xiaoke Hu
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  3. Sean Cook
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  4. Maria Begonia
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Correspondence to Huey-Min Hwang.

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Wang, P., Hu, X., Cook, S. et al. Effect of culture conditions on the production of ligninolytic enzymes by white rot fungi Phanerochaete chrysosporium (ATCC 20696) and separation of its lignin peroxidase. World J Microbiol Biotechnol 24, 2205–2212 (2008). https://doi.org/10.1007/s11274-008-9731-5

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  • DOI: https://doi.org/10.1007/s11274-008-9731-5

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