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A Simple Technique of Preparing Stable CLEAs of Phenylalanine Ammonia Lyase Using Co-aggregation with Starch and Bovine Serum Albumin

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Abstract

Cross-linked enzyme aggregates (CLEAs) have been recently proposed as an alternative to conventional immobilization methods on solid carriers. However, the low cross-linking efficiency causes the major activity loss and instability in the conventional protocol for CLEA preparation. Herein, the effects of bovine serum albumin and starch addition on the cross-linking efficiency of CLEAs of phenylalanine ammonia lyase (PAL) from Rhodotorula glutinis were evaluated. A co-aggregation strategy was developed to improve cross-linking efficiency by adding starch and bovine serum albumin (BSA). CLEAs of PAL prepared in the presence of BSA and starch (PSB-CLEAs) retained 36 % activity, whereas CLEAs prepared without BSA and starch (PAL-CLEAs) retained only 8 % activity of the starting enzyme preparation. Compared with PAL-CLEAs, the thermal stability of PSB-CLEAs has improved considerably, maintaining 30 % residual activity after 4 h of incubation at 70 °C, whereas the PAL-CLEAs have only 13 % residual activity. PSB-CLEAs also exhibited the expected increased stability of PAL against hydrophilic organic solvents, superior operability, and higher storage stability. The proposed technique of preparing CLEAs using co-aggregation with starch and BSA would rank among the potential strategies for efficiently preparing robust and highly stable enzyme aggregates.

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Acknowledgments

The project was partially supported by the National Natural Science Foundation of China (project no. 21072041), Open Funding Project of the National Key Laboratory of Biochemical Engineering (no. KF2010-12) and the Foundation (no. 2012IM004) of Tianjin Key Laboratory of Industrial Microbiology (Tianjin University of Science and Technology), People’s Republic of China, and the Foundation of Hebei University of Science and Technology for Distinguished Young Scientists.

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Authors and Affiliations

  1. Research Center for Fermentation Engineering of Hebei, College of Bioscience and Bioengineering, Hebei University of Science and Technology, 26 YuXiang Street, Shijiazhang, 050000, People’s Republic of China

    Jian Dong Cui, Li Mei Sun & Lian Lian Li

  2. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Haidian District, Beijing, 100080, People’s Republic of China

    Jian Dong Cui

  3. Key Laboratory of Industry Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, 29 Thirteenth Street, Tianjin, 300457, People’s Republic of China

    Jian Dong Cui

  4. College of Bioscience and Bioengineering, Hebei University of Science and Technology, 26 YuXiang Street, Shijiazhuang, 050018, People’s Republic of China

    Jian Dong Cui

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  1. Jian Dong Cui
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  2. Li Mei Sun
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  3. Lian Lian Li
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Correspondence to Jian Dong Cui.

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Cui, J.D., Sun, L.M. & Li, L.L. A Simple Technique of Preparing Stable CLEAs of Phenylalanine Ammonia Lyase Using Co-aggregation with Starch and Bovine Serum Albumin. Appl Biochem Biotechnol 170, 1827–1837 (2013). https://doi.org/10.1007/s12010-013-0317-9

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  • DOI: https://doi.org/10.1007/s12010-013-0317-9

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