Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri
Abstract Background The enzyme phenylalanine ammonia lyase (PAL) controls the transition from primary to secondary metabolism by converting L-phenylalanine (L-Phe) to cinnamic acid. However, the function of PAL in pear plants (Pyrus bretschneideri) has not yet been fully elucidated. Results We ident...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-12-01
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| Series: | BMC Plant Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12870-023-04586-0 |
| _version_ | 1797415658147807232 |
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| author | Guohui Li Cheng Song Muhammad Aamir Manzoor Daoyuan Li Yunpeng Cao Yongping Cai |
| author_facet | Guohui Li Cheng Song Muhammad Aamir Manzoor Daoyuan Li Yunpeng Cao Yongping Cai |
| author_sort | Guohui Li |
| collection | DOAJ |
| description | Abstract Background The enzyme phenylalanine ammonia lyase (PAL) controls the transition from primary to secondary metabolism by converting L-phenylalanine (L-Phe) to cinnamic acid. However, the function of PAL in pear plants (Pyrus bretschneideri) has not yet been fully elucidated. Results We identified three PAL genes (PbPAL1, PbPAL2 and PbPAL3) from the pear genome by exploring pear genome databases. The evolutionary tree revealed that three PbPALs were classified into one group. We expressed PbPAL1 and PbPAL2 recombinant proteins, and the purified PbPAL1 and PbPAL2 proteins showed strict substrate specificity for L-Phe, no activity toward L-Tyr in vitro, and modest changes in kinetics and enzyme characteristics. Furthermore, overexpression of PbAL1 and PbPAL1-RNAi, respectively, and resulted in significant changes in stone cell and lignin contents in pear fruits. The results of yeast one-hybrid (Y1H) assays that PbWLIM1 could bind to the conserved PAL box in the PbPAL promoter and regulate the transcription level of PbPAL2. Conclusions Our findings not only showed PbPAL’s potential role in lignin biosynthesis but also laid the foundation for future studies on the regulation of lignin synthesis and stone cell development in pear fruit utilizing molecular biology approaches. |
| first_indexed | 2024-03-09T05:51:45Z |
| format | Article |
| id | doaj.art-ea0e42b6c7b045bda33f570f4dfb2199 |
| institution | Directory Open Access Journal |
| issn | 1471-2229 |
| language | English |
| last_indexed | 2024-03-09T05:51:45Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Plant Biology |
| spelling | doaj.art-ea0e42b6c7b045bda33f570f4dfb21992023-12-03T12:17:11ZengBMCBMC Plant Biology1471-22292023-12-0123111410.1186/s12870-023-04586-0Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideriGuohui Li0Cheng Song1Muhammad Aamir Manzoor2Daoyuan Li3Yunpeng Cao4Yongping Cai5Anhui Engineering Research Center for Eco-Agriculture of Traditional Chinese Medicine, Anhui Provincial Key Laboratory for Quality Evaluation and Improvement of Traditional Chinese Medicine, College of Biological and Pharmaceutical Engineering, West Anhui UniversityAnhui Engineering Research Center for Eco-Agriculture of Traditional Chinese Medicine, Anhui Provincial Key Laboratory for Quality Evaluation and Improvement of Traditional Chinese Medicine, College of Biological and Pharmaceutical Engineering, West Anhui UniversityDepartment of Plant Science, School of Agriculture and Biology, Shanghai Jiao Tong UniversityAnhui Engineering Research Center for Eco-Agriculture of Traditional Chinese Medicine, Anhui Provincial Key Laboratory for Quality Evaluation and Improvement of Traditional Chinese Medicine, College of Biological and Pharmaceutical Engineering, West Anhui UniversityCAS Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of SciencesAnhui Agricultural UniversityAbstract Background The enzyme phenylalanine ammonia lyase (PAL) controls the transition from primary to secondary metabolism by converting L-phenylalanine (L-Phe) to cinnamic acid. However, the function of PAL in pear plants (Pyrus bretschneideri) has not yet been fully elucidated. Results We identified three PAL genes (PbPAL1, PbPAL2 and PbPAL3) from the pear genome by exploring pear genome databases. The evolutionary tree revealed that three PbPALs were classified into one group. We expressed PbPAL1 and PbPAL2 recombinant proteins, and the purified PbPAL1 and PbPAL2 proteins showed strict substrate specificity for L-Phe, no activity toward L-Tyr in vitro, and modest changes in kinetics and enzyme characteristics. Furthermore, overexpression of PbAL1 and PbPAL1-RNAi, respectively, and resulted in significant changes in stone cell and lignin contents in pear fruits. The results of yeast one-hybrid (Y1H) assays that PbWLIM1 could bind to the conserved PAL box in the PbPAL promoter and regulate the transcription level of PbPAL2. Conclusions Our findings not only showed PbPAL’s potential role in lignin biosynthesis but also laid the foundation for future studies on the regulation of lignin synthesis and stone cell development in pear fruit utilizing molecular biology approaches.https://doi.org/10.1186/s12870-023-04586-0Phenylalanine ammonia lyase (PAL)LigninKineticsPyrus bretschneideri |
| spellingShingle | Guohui Li Cheng Song Muhammad Aamir Manzoor Daoyuan Li Yunpeng Cao Yongping Cai Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri BMC Plant Biology Phenylalanine ammonia lyase (PAL) Lignin Kinetics Pyrus bretschneideri |
| title | Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri |
| title_full | Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri |
| title_fullStr | Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri |
| title_full_unstemmed | Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri |
| title_short | Functional and kinetics of two efficient phenylalanine ammonia lyase from Pyrus bretschneideri |
| title_sort | functional and kinetics of two efficient phenylalanine ammonia lyase from pyrus bretschneideri |
| topic | Phenylalanine ammonia lyase (PAL) Lignin Kinetics Pyrus bretschneideri |
| url | https://doi.org/10.1186/s12870-023-04586-0 |
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