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Biochemical Characterization of Novel Phenylalanine Ammonia-Lyase from Spirulina CPCC-695

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Abstract

Phenylalanine ammonia lyase (PAL) catalyzes the deamination of phenylalanine to cinnamic acid and ammonia. It plays a crucial role in the formation of secondary metabolites through the phenylpropanoid pathway. Recently there has been growing interest in exploring the biochemical properties of PAL for its clinical and commercial applications. PAL as a key component has been used in metabolic engineering and synthetic biology. Due to its high substrate specificity and catalytic efficacy, PAL has opened a new area of interest in the biomedical field. PAL has been frequently used in the enzyme replacement therapy of phenylketonuria, cancer treatment and microbial production of l-phe the precursor of noncalorific sweetener aspartame (Methyl l-α-aspartyl-l-phenylalaninate), antimicrobial and health supplements. PAL occurs in few plants, fungi, bacteria, and cyanobacteria. The present investigation is a preliminary study in which an attempt has been made for the isolation, partial purification, and biochemical characterization of PAL (crude and partially purified) from Spirulina CPCC-695. Partially purified PAL exhibited higher enzymatic activity and protein content than the crude enzyme. Molecular weight of the crude and partially purified PAL was ~ 66 kDa. The optimum temperature and pH for PAL activity was observed as 30 ℃ and 8.0 respectively. l-Phe was the most preferred substrate (100 mM) whereas gallic acid showed maximum inhibition of PAL activity. Enzyme kinetics suggested good catalytic efficacy of the PAL enzyme and affinity towards substrate. Both the enzyme (crude and partially purified) showed less than 5% haemolysis suggesting the biocompatible nature of PAL.

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Acknowledgements

Ahmad. R. sincerely thanks University Grants Commission (F.No. 61-1/2019 (SA-III)), India, for providing Maulana Azad National Fellowship (MANF-SRF). Authors are thankful to Culture Collection Centres of India, University of Madras, for providing the Spirulina CPCC-695 species.

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Authors and Affiliations

  1. Cyanobacterial Biotechnology Laboratory, Department of Biosciences, Jamia Millia Islamia, Central University, New Delhi, 110025, India

    Rakhshan Ahmad, Neha Sami, Gulnar Perveen & Tasneem Fatma

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  1. Rakhshan Ahmad
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  2. Neha Sami
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  4. Tasneem Fatma
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RA and TF designed the experiment, NS helped in the experiments and data curation. RA and GP carried out the lab work. RA wrote the initial manuscript that was corrected and finalised by TF.

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Correspondence to Tasneem Fatma.

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Ahmad, R., Sami, N., Perveen, G. et al. Biochemical Characterization of Novel Phenylalanine Ammonia-Lyase from Spirulina CPCC-695. Protein J 41, 414–423 (2022). https://doi.org/10.1007/s10930-022-10063-8

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