Abstract
Phenylalanine ammonia lyase (PAL, EC 4.3.1.5) is the first committed limiting enzyme of phenylpropanoid pathway which catalyzes the deamination of phenylalanine to cinnamic acid. In the present study, we have cloned, functionally characterized and studied the expression pattern of phenylalanine ammonia lyase (PAL) involved in lignification of secondary cell wall. Phenylalanine ammonia lyase gene was cloned using RACE protocol from Erianthus arundinaceus (EaPAL). The 3.2-kb fragment consists of EaPAL with an ORF of 2116 bp encoding for 705 amino acid residues flanked by 172 bp of 5’ untranslated region and 300 bp of 3’ untranslated region. The tertiary structure of EaPAL showed 57% α-helixes, 3% β-sheet and 18% random coil regions with a calculated pI and molecular weight of 6.31 and ~ 77 KDa, respectively. The SDS-PAGE analysis showed that PAL protein was expressed at 1 mM IPTG induction at ~ 76 kDa. The expression profile of PAL across different tissues (leaf, stem and root) of a hybrid (Co 86032) and Erianthus (IK76-81) showed 2.5-fold increased expression in stem and root tissues of both the clones. Therefore, the genetic information, gene structure and expression of EaPAL might be used for achieving improved biofuel feedstocks.
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Acknowledgements
The authors wish to thank Dr. G. Hemaprabha, Head, Division of Crop Improvement, ICAR Sugarcane Breeding Institute, Coimbatore, for kindly providing sugarcane clones.
Funding
This work was financially supported by Indian Council of research (ICAR SBI P1-15/1–4: (V)).
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Authors Lakshmi Kasirajan contributed to the study conception and design. Material preparation, data collection and analysis were performed by Lakshmi Kasirajan, Rabisha Valiyaparambth and Keerthana Kamaraj. Gadha Ajith, Jijo Joise, Aathisivan Kubandiran and Janani Velu performed sequence analysis. The first draft of the manuscript was written by LK, RV, KK and KK, and LK critically revised the manuscript. All authors read and approved the final manuscript.
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Kasirajan, L., Valiyaparambth, R., Kamaraj, K. et al. Full-Length Cloning, Expression and Functional Characterization of Phenylalanine Ammonia Lyase (PAL) Involved in Lignification of Secondary Cell Wall of Erianthus arundinaceus. Sugar Tech 24, 1728–1737 (2022). https://doi.org/10.1007/s12355-022-01147-x
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DOI: https://doi.org/10.1007/s12355-022-01147-x