Engineered Zea mays phenylalanine ammonia-lyase for improve the catalytic efficiency of biosynthesis trans-cinnamic acid and p-coumaric acid

Zheng, Jiangmei; Sun, Ruobin; Wu, Dan; Chen, Pengcheng; Zheng, Pu

doi:10.1016/j.enzmictec.2024.110423

Cited by 1 publication

(1 citation statement)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…pSUFER was employed for the identification of suboptimal residues, as described by Sun et al, , and the dissimilarity in energy levels between the refined configuration and the solitary point mutant was computed by utilizing the ligand-free Bb Myr structure (PDB: 1WCG) as the initial file. Sites with numerous mutations in ΔΔ G ≤ 0 R.e.u.…”

Section: Methodsmentioning

confidence: 99%

Computer-driven Evolution of Myrosinase from the Cabbage Aphid for Efficient Production of (R)-Sulforaphane

Sun,

Huang,

Wang

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

Myrosinase (Myr) catalyzes the hydrolysis of glucosinolates, yielding biologically active metabolites. In this study, glucoraphanin (GRA) extracted from broccoli seeds was effectively hydrolyzed using a Myr-obtained cabbage aphid (Brevicoryne brassicae) (BbMyr) to produce (R)-sulforaphane (SFN). The gene encoding BbMyr was successfully heterologously expressed in Escherichia coli, resulting in the production of 1.6 g/L (R)-SFN, with a remarkable yield of 20.8 mg/g broccoli seeds , achieved using recombination E. coli whole-cell catalysis under optimal conditions (pH 4.5, 45 °C). Subsequently, BbMyr underwent combinatorial simulation-driven mutagenesis, yielding a mutant, DE9 (N321D/Y426S), showing a remarkable 2.91-fold increase in the catalytic efficiency (k cat /K M ) compared with the original enzyme. Molecular dynamics simulations demonstrated that the N321D mutation in loopA of mutant DE9 enhanced loopA stability by inducing favorable alterations in hydrogen bonds, while the Y426S mutation in loopB decreased spatial resistance. This research lays a foundation for the environmentally sustainable enzymatic (R)-SFN synthesis.

show abstract

Section: Methodsmentioning

confidence: 99%