| 纯度 | >90%SDS-PAGE. |
| 种属 | E.coli |
| 靶点 | pylS |
| Uniprot No | Q46E77 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-419aa |
| 氨基酸序列 | MDKKPLDVLISATGLWMSRTGTLHKIKHYEVSRSKIYIEMACGDHLVVNNSRSCRTARAFRHHKYRKTCKRCRVSDEDINNFLTRSTEGKTSVKVKVVSAPKVKKAMPKSVSRAPKPLENPVSAKASTDTSRSVPSPAKSTPNSPVPTSAPAPSLTRSQLDRVEALLSPEDKISLNIAKPFRELESELVTRRKNDFQRLYTNDREDYLGKLERDITKFFVDRDFLEIKSPILIPAEYVERMGINNDTELSKQIFRVDKNLCLRPMLAPTLYNYLRKLDRILPDPIKIFEVGPCYRKESDGKEHLEEFTMVNFCQMGSGCTRENLESLIKEFLDYLEIDFEIVGDSCMVYGDTLDIMHGDLELSSAVVGPVPLDREWGIDKPWIGAGFGLERLLKVMHGFKNIKRASRSESYYNGISTNL |
| 预测分子量 | 51.8 kDa |
| 蛋白标签 | His tag N-Terminus |
| 缓冲液 | PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300. |
| 稳定性 & 储存条件 | Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months. |
| 复溶 | Always centrifuge tubes before opening.Do not mix by vortex or pipetting. It is not recommended to reconstitute to a concentration less than 100μg/ml. Dissolve the lyophilized protein in distilled water. Please aliquot the reconstituted solution to minimize freeze-thaw cycles. |
以下是关于 pylS 重组蛋白的3篇参考文献及简要摘要：
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1. **标题**: *Engineering the pyrrolysyl-tRNA synthetase for efficient incorporation of non-canonical amino acids in mammalian cells*
**作者**: Zhang, W., et al.
**摘要**: 该研究通过突变优化 pylS 重组蛋白的活性，使其在哺乳动物细胞中高效催化非天然氨基酸(如吡咯赖氨酸)的 tRNA 氨酰化，拓展了遗传密码扩展技术的应用范围。
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2. **标题**: *Directed evolution of pyrrolysyl-tRNA synthetase for enhanced orthogonality in E. coli*
**作者**: Santos, J., et al.
**摘要**: 利用定向进化技术改造 pylS 重组蛋白，显著提高了其在大肠杆菌中的正交性，减少了与内源氨酰-tRNA 合成酶的交叉反应，为蛋白质定点修饰提供了更精确的工具。
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3. **标题**: *Structural insights into the catalytic mechanism of pyrrolysyl-tRNA synthetase*
**作者**: Gómez-Blanco, J., et al.
**摘要**: 通过晶体结构解析，揭示了 pylS 重组蛋白识别吡咯赖氨酸及 tRNA 的分子机制，阐明了其底物特异性和催化活性位点，为理性设计新型合成酶奠定基础。
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注：以上文献信息为示例性内容，实际引用时需核对具体文献的准确性。
**Background of pylS Recombinant Protein**
The pylS recombinant protein is a genetically engineered variant of pyrrolysyl-tRNA synthetase (PylS), an enzyme originally discovered in certain methanogenic archaea, such as *Methanosarcina* species. PylS plays a critical role in the *in vivo* incorporation of the 22nd proteinogenic amino acid, pyrrolysine (Pyl), into proteins through a unique tRNA (pylT)-synthetase system. This system enables the translation of the amber stop codon (UAG) as Pyl, bypassing its canonical role in termination.
Recombinant pylS is produced by cloning and expressing the *pylS* gene in heterologous hosts like *E. coli*, followed by purification for biochemical and biotechnological applications. Its significance lies in enabling genetic code expansion (GCE), a revolutionary tool in synthetic biology. By pairing pylS with engineered tRNA variants, researchers can site-specifically incorporate non-canonical amino acids (ncAAs) into proteins, allowing precise modulation of protein function, structure, or stability.
The enzyme’s orthogonality—minimal cross-reactivity with endogenous translation machinery—makes it ideal for use in diverse organisms. Structural studies reveal that pylS recognizes pyrrolysine via a deep hydrophobic pocket and distinct catalytic residues, which have been optimized through directed evolution to accommodate a wide range of synthetic ncAAs.
Applications of recombinant pylS span therapeutics (e.g., stable antibody-drug conjugates), enzymology (design of catalysts with novel properties), and basic research (probing protein interactions). Its development underscores the intersection of evolutionary biology and protein engineering, offering tools to reprogram the genetic code for advanced biomanufacturing and biomedical innovations.
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