Protein engineering provides a powerful base for the circumvention of challenges tied with characteristics accountable for enzyme functions. CYP82Y1 introduces a hydroxyl group (−OH) into C1 of N-methylcanadine as the substrate to yield 1-hydroxy-N-methylcanadine. This chemical process has been found to be the gateway to noscapine biosynthesis. Owning to the importance of CYP82Y1 in this biosynthetic pathway, it has been selected as a target for enzyme engineering. The insertion of tags to the N- and C-terminal of CYP82Y1 was assessed for their efficiencies for improvement of the physiological performances of CYP82Y1. Although these attempts achieved some positive results, further strategies are required to dramatically enhance the CYP82Y1 activity. Here methods that have been adopted to achieve a functionally improved CYP82Y1 will be reviewed. In addition, the possibility of recruitment of other techniques having not yet been implemented in CYP82Y1 engineering, including the substitution of the residues located in the substrate recognition site, formation of the synthetic fusion proteins, and construction of the artificial lipid-based scaffold will be discussed. Given the fact that the pace of noscapine synthesis is constrained by the CYP82Y1-catalyzing step, the methods proposed here are capable of accelerating the rate of reaction performed by CYP82Y1 through improving its properties, resulting in the enhancement of noscapine accumulation.

中文翻译：

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细胞色素 P450 单加氧酶 CYP82Y1 的工程改造：罂粟中诺斯卡品生物合成的关键酶

蛋白质工程为规避与酶功能相关的特征相关的挑战提供了强大的基础。CYP82Y1 将羟基（-OH）引入到作为底物的 N-甲基卡纳定的 C1 中，产生 1-羟基-N-甲基卡纳定。已发现该化学过程是那可品生物合成的途径。由于 CYP82Y1 在该生物合成途径中的重要性，它已被选为酶工程的靶标。评估了将标签插入 CYP82Y1 N 端和 C 端对改善 CYP82Y1 生理性能的效率。尽管这些尝试取得了一些积极的成果，但仍需要进一步的策略来显着增强 CYP82Y1 活性。此处将回顾用于实现功能改进的 CYP82Y1 所采用的方法。此外，招募尚未在 CYP82Y1 工程中实施的其他技术的可能性，包括位于底物识别位点的残基的取代、合成融合蛋白的形成以及人工脂质支架的构建将是讨论过。鉴于那斯卡品的合成速度受到 CYP82Y1 催化步骤的限制，本文提出的方法能够通过改善 CYP82Y1 的性能来加速 CYP82Y1 的反应速率，从而增强那斯卡品的积累。