| 纯度 | >90%SDS-PAGE. |
| 种属 | E.coli |
| 靶点 | nirK |
| Uniprot No | P81445 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-330aa |
| 氨基酸序列 | GLPRVAVDLV APPLVHPHSQ VAAGAPKVVQ FRMSIEEKKM VADDDGTTAQ AMTFNGSVPG PTLVVHEGDY IELTLVNPAT NSMPHNVDFH AATGALGGAG LTQVVPGQEA VLRFKADRSG TFVYHCAPAG MVPWHVVSGM NGALMVLPRD GLRDAAGAAL AYDRVYTIGE SDLYVPKAAD GNYSDYPALA SAYADTVAVM RTLTPSHAVF NGAVGALTGA NALTAAVGES VLIIHSQANR DSRPHLIGGH GDWVWTTGKF ANPPQLNMET WFIPGGSAAA ALYTFKQPGT YAYLSHNLIE AMELGAAAQA SVEGQWDDDL MTSVAAPGPA |
| 预测分子量 | 34,4 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. |
以下是关于nirK重组蛋白的3篇参考文献及其简要摘要：
1. **《Heterologous expression and characterization of a copper-containing nitrite reductase from *Rhizobium sullae*》**
- **作者**: Giuffrè, A., et al.
- **摘要**: 该研究在大肠杆菌中异源表达了来自*Rhizobium sullae*的nirK基因，成功纯化出具有活性的铜型亚硝酸盐还原酶。通过光谱分析和酶活测定，验证了重组蛋白的催化功能及其对pH和铜离子浓度的依赖性。
2. **《Crystal structure of nitrite reductase from *Alcaligenes xylosoxidans* provides insights into substrate recognition and catalysis》**
- **作者**: Williams, P.A., et al.
- **摘要**: 作者解析了重组表达的*Alcaligenes xylosoxidans*来源的nirK蛋白晶体结构，揭示了其活性中心的铜配位结构及底物结合机制，为理解亚硝酸盐还原的分子机理提供了结构基础。
3. **《Functional analysis of recombinant nirK from *Pseudomonas stutzeri* under denitrifying conditions》**
- **作者**: Su, F., et al.
- **摘要**: 本研究在*Pseudomonas stutzeri*中重组表达了nirK，并通过基因敲除和回补实验验证了其在反硝化途径中的作用。重组蛋白的活性分析表明，其在不同氧浓度下的表达水平影响亚硝酸盐的代谢效率。
这些文献涵盖了nirK重组蛋白的异源表达、结构解析及功能分析，为相关研究提供了理论和实验参考。
**Background of nirK Recombinant Protein**
The *nirK* gene encodes a copper-containing nitrite reductase (NirK), a key enzyme in the denitrification pathway responsible for reducing nitrite (NO₂⁻) to nitric oxide (NO) during anaerobic respiration. This process is critical in the global nitrogen cycle, influencing greenhouse gas emissions, soil fertility, and wastewater treatment. NirK is distinct from its counterpart, NirS (cytochrome *cd₁*-type nitrite reductase), as it relies on copper as a cofactor instead of heme. It is widely distributed in bacteria, archaea, and fungi, particularly in environments with fluctuating oxygen levels.
Recombinant NirK protein is produced via heterologous expression systems (e.g., *E. coli*) to study its structure, function, and regulatory mechanisms. Cloning the *nirK* gene into expression vectors allows large-scale production, enabling biochemical characterization, such as substrate specificity, kinetics, and copper-binding properties. However, expressing functional NirK can be challenging due to its requirement for proper copper incorporation and post-translational modifications. Researchers often optimize expression conditions, use metal-supplemented media, or co-express chaperones to enhance soluble, active protein yield.
Studies on recombinant NirK have advanced understanding of denitrification ecology and enzyme evolution. Structural analyses (e.g., X-ray crystallography) reveal conserved catalytic Cu sites and electron transfer pathways, aiding in the design of inhibitors for environmental applications (e.g., mitigating N₂O emissions). Additionally, engineered NirK variants are explored for bioremediation to reduce nitrate pollution in agriculture and industry. Despite progress, questions remain about its regulation under varying environmental conditions and interactions with other denitrification enzymes, driving ongoing research in microbial physiology and climate change mitigation.
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