High-level and reusable preparation of sulforaphane by yeast cells expressing myrosinase

Wang, Lili; Hamouda, Hamed I.; Dong, Yueyang; Jiang, Hong; Quan, Yongyi; Chen, Yimiao; Liu, Yan; Wang, Jiaqi; Balah, Mohamed A.; Mao, Xiangzhao

doi:10.1016/j.fochx.2023.100668

Cited by 1 publication

(1 citation statement)

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“…The addition of exogenous Myr into GRA-containing plant extracts increased the SFN yield. However, almost all commercial Myr is extracted from white Sinapis alba seeds with complex preparation procedures, and these natural Myrs do not exhibit high hydrolytic activity against GRA . Due to the drawbacks associated with isolating and purifying Myr extracted from natural plants and bacteria, which often lead to increased costs, utilizing heterologous expression hosts for the production of Myr has emerged as an effective alternative.…”

Section: Introductionmentioning

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.

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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.

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Section: Introductionmentioning

confidence: 99%