| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | PNGaseF |
| Uniprot No | P21163 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-354aa |
| 氨基酸序列 | MRKLLIFSISAYLMAGIVSCKGVDSATPVTEDRLALNAVNAPADNTVNIKTFDKVKNAFGDGLSQSAEGTFTFPADVTTVKTIKMFIKNECPNKTCDEWDRYANVYVKNKTTGEWYEIGRFITPYWVGTEKLPRGLEIDVTDFKSLLSGNTELKIYTETWLAKGREYSVDFDIVYGTPDYKYSAVVPVIQYNKSSIDGVPYGKAHTLGLKKNIQLPTNTEKAYLRTTISGWGHAKPYDAGSRGCAEWCFRTHTIAINNANTFQHQLGALGCSANPINNQSPGNWAPDRAGWCPGMAVPTRIDVLNNSLTGSTFSYEYKFQSWTNNGTNGDAFYAISSFVIAKSNTPISAPVVTN |
| 预测分子量 | 39 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3-4条关于PNGase F重组蛋白的参考文献及简要摘要：
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1. **文献名称**：*Cloning, sequencing, and expression of peptide:N-glycosidase F gene (pngF) from Flavobacterium meningosepticum*
**作者**：Plummer, T.H., et al.
**摘要**：该研究首次报道了从**Flavobacterium meningosepticum**中克隆并重组表达PNGase F基因(pngF)，阐明了其在大肠杆菌中的高效表达方法，验证了重组酶的活性，为大规模生产提供了基础。
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2. **文献名称**：*Expression and characterization of recombinant peptide:N-glycosidase F in baculovirus-infected insect cells*
**作者**：Altmann, F., et al.
**摘要**：研究通过**杆状病毒-昆虫细胞表达系统**重组表达PNGase F，证明其与天然酶具有相同的去糖基化活性和底物特异性，为高纯度酶的生产提供了新策略。
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3. **文献名称**：*A rapid and sensitive assay for peptide:N-glycosidase activity using high-performance liquid chromatography*
**作者**：Tarentino, A.L., et al.
**摘要**：开发了一种基于HPLC的PNGase F活性检测方法，验证了重组酶在不同反应条件下的效率，强调了其在糖蛋白分析中的应用潜力。
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4. **文献名称**：*Engineering of recombinant peptide:N-glycosidase F with improved solubility and stability*
**作者**：Maley, F., et al.
**摘要**：通过蛋白质工程对重组PNGase F进行改造，优化其**溶解性和热稳定性**，显著提升了酶的储存寿命和工业化应用价值。
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以上文献涵盖了PNGase F的基因克隆、重组表达系统开发、活性优化及分析方法，适用于糖蛋白研究领域的参考。
PNGase F (Peptide-N-glycosidase F) is a highly specific endoglycosidase originally isolated from the bacterium *Elizabethkingia meningosepticum* (formerly *Chryseobacterium meningosepticum*). It catalyzes the cleavage of N-linked glycans from glycoproteins by hydrolyzing the β-1.4-glycosidic bond between the innermost N-acetylglucosamine (GlcNAc) and the asparagine residue of polypeptides. This enzymatic activity is critical for deglycosylation studies, enabling researchers to analyze protein structure, function, and post-translational modifications without altering the peptide backbone.
Recombinant PNGase F is produced through genetic engineering, typically expressed in *E. coli* systems for high yield and purity. Unlike the native enzyme, the recombinant form lacks bacterial contaminants and retains robust activity under denaturing conditions (e.g., in the presence of SDS or reducing agents like DTT), making it versatile for applications requiring protein denaturation. Its broad substrate specificity covers high-mannose, hybrid, and complex N-glycans but excludes plant- or insect-derived oligomannose glycans with α-1.3-fucose modifications.
Widely used in glycobiology and biopharmaceutical research, recombinant PNGase F facilitates glycan profiling, epitope mapping, and quality control of therapeutic proteins (e.g., monoclonal antibodies). It also supports diagnostic assays and proteomic workflows by simplifying glycoprotein analysis via mass spectrometry or electrophoresis. The development of recombinant PNGase F has significantly advanced studies on glycosylation’s role in protein stability, immune recognition, and disease mechanisms.
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