| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | DHDH |
| Uniprot No | Q9UQ10 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-334aa |
| 氨基酸序列 | MALRWGIVSVGLISSDFTAVLQTLPRSEHQVVAVAARDLSRAKEFAQKHDIPKAYGSYEELAKDPSVEVAYIGTQHPQHKAAVMLCLAAGKAVLCEKPTGVNAAEVREMVAEARSRALFLMEAIWTRFFPASEALRSVLAQGTLGDLRVARAEFGKNLIHVPRAVDRAQAGGALLDIGIYCVQFTSMVFGGQKPEKISVVGRRHETGVDDTVTVLLQYPGEVHGSFTCSITVQLSNTASVSGTKGMVQLLNPCWCPTELVVKGEHKEFPLPPVPKDCNFDNGAGMSYEAKHVWECLRKGMKESPVIPLSESELLADILEEVRKAIGVTFPQDKR |
| 预测分子量 | 63.4 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3篇与DHDH(假设为二羟基酸脱氢酶)重组蛋白相关的文献示例，基于常见研究方向虚构，供参考：
1. **标题**: "Heterologous expression and characterization of recombinant DHDH from Bacillus subtilis in E. coli"
**作者**: Li, X.; Wang, H.; Zhang, Y.
**摘要**: 本研究成功在大肠杆菌中克隆表达了枯草芽孢杆菌来源的DHDH酶，优化了表达条件(IPTG浓度、温度)，纯化后酶比活达到120 U/mg，并验证了其在支链氨基酸合成中的催化功能。
2. **标题**: "Structural insights into the catalytic mechanism of DHDH through X-ray crystallography"
**作者**: Smith, J.; Tanaka, K.; et al.
**摘要**: 通过X射线晶体学解析了重组人源DHDH的2.1Å分辨率三维结构，揭示了其底物结合口袋的关键氨基酸残基(Arg158和Glu202)，为设计特异性抑制剂提供结构基础。
3. **标题**: "Application of recombinant DHDH in the biosynthesis of branched-chain alcohols"
**作者**: Chen, L.; Park, S.; et al.
**摘要**: 将重组DHDH与丙酮酸脱羧酶偶联，构建体外催化体系，实现了α-酮异己酸高效转化为异丁醇(产率85%)，展示了其在生物燃料合成中的潜力。
注：以上文献为模拟示例，实际研究中建议通过PubMed或Web of Science检索关键词"DHDH recombinant protein"或"Dihydroxyacid dehydrogenase expression"获取真实文献。
Dihydroxyacid dehydrogenase (DHDH), a key enzyme in the branched-chain amino acid (BCAA) biosynthesis pathway, catalyzes the oxidative decarboxylation of α,β-dihydroxy acids to yield α-keto acids. This NAD⁺-dependent enzyme plays a critical role in producing essential amino acids (e.g., valine, leucine, isoleucine) and derivatives used in metabolic processes. Its conserved structure across species – typically a homodimer with Rossmann-fold domains for cofactor binding – makes it a model for studying enzyme mechanisms and substrate specificity.
Recombinant DHDH technology emerged to address challenges in native protein extraction, such as low yield and purity from natural sources. By cloning the DHDH gene (e.g., ilvD in E. coli) into expression vectors (e.g., pET or pQE systems), researchers achieve high-level production in host systems like E. coli or yeast. Optimization of codon usage, induction conditions (e.g., IPTG concentration), and purification tags (His-tag, GST) enhances soluble expression and simplifies affinity chromatography purification.
Applications span multiple fields:
1. **Biocatalysis**: Engineered DHDH variants enable efficient synthesis of chiral intermediates for pharmaceuticals and agrochemicals.
2. **Metabolic engineering**: Overexpression in microbial hosts improves BCAA production for nutritional supplements.
3. **Antibiotic research**: As DHDH is absent in mammals but vital for bacterial/fungal metabolism, it serves as a target for developing novel antimicrobials.
4. **Enzyme kinetics studies**: Recombinant protein facilitates structure-function analysis through mutagenesis and crystallography.
Recent advances include fusion protein strategies to enhance stability and immobilization techniques for industrial biocatalysis. Challenges remain in balancing enzyme activity with heterologous expression efficiency, particularly for thermophilic DHDH variants. Ongoing research focuses on directed evolution and computational design to expand substrate range and improve industrial applicability.
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