Coding

Part:BBa_K5218001

Designed by: Rui Zhang, Lichuan Chen, Xiaojuan Wang   Group: iGEM24_BGI-MammothEdu-South   (2024-08-10)


RcTAL

Tyrosine ammonia lyase (TAL) gene from Rhodobacter capsulatus

Base Pairs：1599 bp

Function：Tyrosine ammonia-lyase (TAL) that catalyzes the formation of p-coumaric acid from tyrosine.

Figure.1: TAL plays a role in the pathway of caffeic acid biosynthesis.
Figure adopted from L. Liu et al, 2019 [1]

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 490
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 490
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 490
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 490
    Illegal AgeI site found at 646
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

Ammonia-lyases catalyze the deamination of amino acids. Members of this family include histidine ammonia lyase (HAL) that converts histidine to urocanic acid, phenylalanine ammonia lyase (PAL) that converts phenylalanine to cinnamate (CA), and tyrosine ammonia lyase (TAL) that converts tyrosine to p-coumaric acid. p-coumaric acid is an important precursor in many metabolic pathways, one of which is the production of caffeic acid with the catalysis of coumarate 3-hydroxylase (C3H).

PALs, as key rate-limiting enzymes, are commonly found in plants, algae, fungi and bacteria[2]. The presence of TAL enzymes have been reported in a few microorganisms. TAL from Rhodosporidium toruloides was demonstrated to synthesize p-coumaric acid from L-tyrosine and obtained a yield of 117.5 mg/L [1]. TAL from Rhodobacter capsulatus has 32% identity with plant PAL sequence of Pinus taeda[3]. iGEM13_Uppsala team registered a TAL gene from rhodobacter sphaeroides (Part:BBa_K1033000) and characterized the enzyme with spectrophotometry and chromatography.

Rhodobacter capsulatus is a species of purple bacteria, a group of bacteria that can obtain energy through photosynthesis(Wikipedia).

The engineering objective of this project is to generate brighter autoluminescent plants using synthetic biology approaches. Team BGI-MammothEdu-South 2024 selected a variety of candidate genes from different species and tested their functions in Fungal Bioluminescence Pathway (FBP) via eGFP reporter system (pS1300-GFP plasmid). Team BGI-MammothEdu-South retrieved the RcTAL CDS from published article[4], and obtained the sequence through De novo DNA synthesis.

Reference

1.Liu, Langqing, Liu, Hong, Zhang Wei, Yao, Mingdong, Li, Bingzhi, Liu, Duo, Yuan Yingjin. (2019) Engineering the biosynthesis of caffeic acid in saccharomyces cerevisiae with heterologous enzyme combinations. Engineering, 5(2), 9.

2.Zhang Fulin, Wang Juan, Li Xianguo, Zhang Jun, Liu Yuxiang , Chen Yijia, Yu Qinghui, Li Ning (2023) Genome-wide identification and expression analyses of phenylalanine ammonia-lyase gene family members from tomato (Solanum lycopersicum) reveal their role in root-knot nematode infection. Frontiers in Plant Science, 14:1204990

3.Kyndt J.A., Meyer T.E., Cusanovich M.A. and Van Beeumen J.J. (2002) Characterization of a bacterial tyrosine ammonia lyase, a biosynthetic enzyme for the photoactive yellow protein, FEBS Letters, 512

4.Arjun Khakhar, Colby G Starker, James C Chamness, Nayoung Lee, Sydney Stokke, Cecily Wang, Ryan Swanson, Furva Rizvi, Takato Imaizumi, Daniel F Voytas (2020) Building customizable auto-luminescent luciferase-based reporters in plants. eLife, 9:e52786

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