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Modification of Aromatic Amino Acid Synthetic Pathway in Pichia pastoris to Produce Cinnamic Acid and ρ-Coumaric Acid |
CHEN Xin-jie1,QIAN Zhi-lan1,LIU Qi1,ZHAO Qing2,ZHANG Yuan-xing1,CAI Meng-hao1,**() |
1 State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, China 2 Plant Science Research Center, Shanghai Chenshan Botanical Garden, Shanghai Key Laboratoryof Plant Functional Genomics and Resources, Shanghai 201602, China |
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Abstract Objective: The Pichia pastoris strain was engineered to heterologously synthesize cinnamic acid and ρ-coumaric acid,which were important intermediates of flavonoid biosynthetic pathway. The biosynthetic pathway of precursors aromatic amino acids was optimized to improve the production capacity of P. pastoris. Methods: Phenylalanine ammonia lyase from Rhodotorula glutinis was expressed in P. pastoris GS115 by ethanol induced artificial transcription system, and the key enzymes or their mutants in the biosynthetic pathway of intracellular aromatic amino acids were overexpressed in the recombinant strain. Results: Heterologous expression of phenylalanine ammonia lyase could convert L-phenylalanine and L-tyrosine produced by P. pastoris into cinnamic acid (38.8 mg/L) and ρ-coumaric acid (34.2 mg/L). Through overexpression of related enzymes, the yields of cinnamic acid and ρ-coumaric acid reached 124.1 mg/L and 302.0 mg/L, respectively. Conclusion: Cinnamic acid and ρ-coumaric acid were successfully synthesized by P. pastoris, and the biosynthetic pathway of intracellular aromatic amino acids was optimized. It shows that P. pastoris has the application potential to produce flavonoids, and it also lays a foundation for the heterologous synthesis of other aromatic amino acid derivatives or plant compounds in P. pastoris.
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Received: 11 June 2021
Published: 08 November 2021
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Corresponding Authors:
Meng-hao CAI
E-mail: cmh022199@ecust.edu.cn
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