| 纯度 | >90%SDS-PAGE. |
| 种属 | E.coli |
| 靶点 | PYC1 |
| Uniprot No | P11154 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 18-470aa |
| 氨基酸序列 | EKNKILVANRGEIPIRIFRTAHELSMQTVAIYSHEDRLSTHKQKADEAYVIGEVGQYTPVGAYLAIDEIISIAQKHQVDFIHPGYGFLSENSEFADKVVKAGITWIGPPAEVIDSVGDKVSARNLAAKANVPTVPGTPGPIETVEEALDFVNEYGYPVIIKAAFGGGGRGMRVVREGDDVADAFQRATSEARTAFGNGTCFVERFLDKPKHIEVQLLADNHGNVVHLFERDCSVQRRHQKVVEVAPAKTLPREVRDAILTDAVKLAKECGYRNAGTAEFLVDNQNRHYFIEINPRIQVEHTITEEITGIDIVAAQIQIAAGASLPQLGLFQDKITTRGFAIQCRITTEDPAKNFQPDTGRIEVYRSAGGNGVRLDGGNAYAGTIISPHYDSMLVKCSCSGSTYEIVRRKMIRALIEFRIRGVKTNIPFLLTLLTNPVFIEGTYWTTFIDDTPQ |
| 预测分子量 | 52.0 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PYC1(丙酮酸羧化酶1)重组蛋白研究的模拟参考文献示例(内容为虚构，仅供格式参考)：
1. **文献名称**：*Heterologous Expression and Characterization of Recombinant PYC1 in Escherichia coli*
**作者**：Zhang L., Wang Y., et al.
**摘要**：本研究成功在大肠杆菌中克隆并表达了来源于酿酒酵母的PYC1基因，通过His标签纯化获得高纯度重组蛋白。酶活测定显示重组PYC1在体外具有催化丙酮酸转化为草酰乙酸的能力，最适反应温度为37℃，为后续代谢工程研究奠定基础。
2. **文献名称**：*Optimization of PYC1 Protein Production in Pichia pastoris for Biotechnological Applications*
**作者**：Müller S., Braun F.
**摘要**：通过毕赤酵母表达系统优化PYC1的分泌表达，采用高密度发酵技术使产量提高5倍。纯化后的蛋白经SDS-PAGE验证为单体形式，动力学分析表明其比活性较原核表达体系提升30%，适用于工业级酶催化反应。
3. **文献名称**：*Structural and Functional Analysis of Pyruvate Carboxylase Isozyme PYC1*
**作者**：Tanaka K., Ito T.
**摘要**：利用X射线晶体学解析了重组PYC1的三维结构(分辨率2.8Å)，发现其活性中心关键氨基酸残基对底物结合的影响。定点突变实验证实Arg278和Glu329在催化中的关键作用，揭示了PYC1与同工酶PYC2的功能差异。
4. **文献名称**：*Role of Recombinant PYC1 in Enhancing Lipid Biosynthesis in Microalgae*
**作者**：Chen H., Li M., et al.
**摘要**：将重组PYC1导入微藻细胞，通过增强草酰乙酸供应使脂肪酸合成效率提高40%。研究证明PYC1过表达可调控碳流向脂质代谢通路，为生物燃料生产提供新策略。
**注意**：以上文献为模拟生成，实际研究中请通过PubMed/Google Scholar检索关键词“pyruvate carboxylase 1 recombinant”、“PYC1 heterologous expression”获取真实文献。
PYC1 (pyruvate carboxylase 1) is a biotin-dependent mitochondrial enzyme that plays a critical role in cellular metabolism, particularly in gluconeogenesis and anaplerotic reactions. Found across eukaryotes, including humans, it catalyzes the ATP-dependent carboxylation of pyruvate to oxaloacetate (OAA), a key intermediate in the tricarboxylic acid (TCA) cycle. This reaction replenishes TCA cycle intermediates and supports biosynthetic pathways in energy-demanding tissues like liver, brain, and adipose tissue. Dysregulation of PYC1 is linked to metabolic disorders, including type II diabetes and rare genetic deficiencies.
Recombinant PYC1 protein is engineered through heterologous expression systems, typically using E. coli or yeast, to study its structure, function, and regulatory mechanisms. Its production enables detailed biochemical characterization, such as substrate binding kinetics, allosteric regulation by acetyl-CoA, and interactions with metabolic partners. Purification often involves affinity tags (e.g., His-tag) followed by size-exclusion chromatography. Research on recombinant PYC1 has advanced understanding of metabolic flux control, insulin secretion pathways in pancreatic β-cells, and mitochondrial dysfunction. It also serves as a tool for drug screening targeting metabolic diseases or cancer, where altered carboxylase activity influences tumor growth. Recent structural studies using recombinant PYC1 (e.g., cryo-EM analyses) have revealed dynamic conformational changes during catalysis, informing therapeutic design. Industrial applications include microbial engineering for biofuel production, leveraging its role in carbon fixation. Despite progress, challenges persist in maintaining its multimeric native structure and enzymatic activity in vitro, driving ongoing optimization of recombinant expression protocols.
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