Bio-protocol
·
2025
·Vol. 15
·期 8
·e5270
·
IF 1.1
Xiaofeng Yang,Zhanglin Lin,Ya Xiang,Binrui Chen,Zisha Lao
Protein purification is a critical step in both life sciences and biomanufacturing. Traditional affinity chromatography (AC) methods, including His-tag-based purification, provide high-purity proteins but are limited by the high cost of resins and the need for additional tag-removal steps. In this protocol, we present a reusable SpyDock-modified epoxy resin coupled with a pH-inducible self-cleaving intein for direct purification of proteins with authentic N-termini. This method enables efficient protein purification from cell lysates, achieving high purity (>90%) and yields comparable to the His-tag approach, without requiring tag removal. The SpyDock-modified resin protocol is robust, easy to implement, and cost-effective, making it suitable for both research and large-scale industrial applications. Key features • This protocol offers a robust and straightforward method for purifying proteins with authentic N-termini, eliminating the need for additional tag removal steps. • The approach achieves higher purity and comparable yields to the commercial His-tag method. • The SpyDock-modified epoxy resin is easy to prepare, cost-effective, and reusable.
研究领域:
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND BIOPRODUCTS PROCESSING
·
2024
·
IF 3.5
Lu Ren,Yang Hu,Yefei Wu,Mengqian Zhao,Qianqian Ma,Yulu Du,Jianwei Lu,Xuwei Long
Sophorose is a rare and valuable disaccharide with an unusual β -1,2 bond. Despite the relatively widespread interest in sophorose, the information regarding sophorose is limited because sophorose is not readily accessible. Thus, we used Candida bombicola ATCC 22214 for the efficient fermentative production of sophorose using glucose and soybean oil as substrates. In addition, an integrated separation process comprising precipitation, extraction, electrodialysis, and column chromatography was developed to purify the sophorose. Sophorose (approximately 35 g/L) was produced under optimized fermentation conditions in terms of the feeding strategy and enhanced mass-transfer efficiency. Thereafter, approximately 38% of sophorose (purity above 99%) was recovered from fermentation broth through the proposed process. This study provides an efficient approach for the large-scale preparation of sophorose, which is helpful for its practical application and in-depth research.
研究领域:
生物工程
·
糖化学
✓ 已验证
产品关联
DOI
高影响因子
Materials Today Energy
·
2024
·
IF 9
Haochen Liu,Xiangtian Xiao,Ye Wu,Ding Zhu,Jiayun Sun,Desui Chen,Bing Tang,Shixun Wang,Arsenii Portniagin,Kunnathodi Vighnesh,Wallace C.H. Choy,Andrey L. Rogach
Pure-red emission light-emitting diodes (LEDs) are highly desirable for display and lighting applications. Here, we present a synergistic approach to fabricate pure-red emission LEDs based on quantum-confined CsPbI 3 nanocrystals (NCs). By introducing ZnI 2 acting as a co-precursor and passivating agent into the synthesis, and applying a gradient post-synthesis purification method allowing us to obtain size-selected fractions with precisely adjusted emission colors, we obtained uniform and small (about 6 nm), quantum-confined CsPbI 3 NCs exhibiting high stability in ambient conditions and a superior photoluminescence quantum yield of up to 88%. These NCs exhibited the tendency to undergo oriented attachment, resulting in self-assembled superstructures with a pure-red emission at 629 nm. Importantly, partial recrystallization into fused superstructures led to removal of the insulating ligand barriers between NCs. LEDs based on the CsPbI 3 NCs showed pure-red electroluminescence at 633 nm, with an external quantum efficiency of 14.7% and a luminance of over 1000 cd/m 2 . Our study reveals the mechanism behind the oriented attachment and assembly of small, quantum-confined CsPbI 3 NCs, and contributes to the development of pure-red electroluminescent LEDs.
研究领域:
材料科学
·
光电技术
·
纳米科技
✓ 已验证
产品关联
DOI
高影响因子
PHYSICA SCRIPTA
·
2025
·Vol. 100
·期 6
·065916
·
IF 2.6
Pouria Pourhosseinhendabad,Parisa Z Firouzabadi
Bentonite clay, nano-bentonite, and nano-clays are used in many industries such as medical, water purification, food processing, and drilling mud. But because impurities in low-grade bentonite occur, its efficiency is limited; therefore, purification and modification are carried out to enhance its quality. Cetyltrimethylammonium bromide (CTAB), cationic modifier, dispersants, and milling are used in this study's improved process for synthesizing nano-bentonite and industrial bentonite purification. CTAB was the preferred choice since it possessed the ability to modify further such parameters as surface charge, hydrophobicity, and cation exchange capacity, thus rendering bentonite more effective for its applications. The removal of impurities was mainly facilitated by the dispersants during sedimentation and centrifugation. The best sample, BMP 2 h, has shown a tremendous reduction in the cristobalite and calcite impurities with an increase in the beidellite phase as confirmed by x-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The process reduced particle size from raw bentonite 121.75 μm to 1.07 μm of BMP 2 h, which enabled improved dispersion. The enhanced purity and changed behavior of nano-bentonite reveal its potential for future industrial use in water treatment, catalysis, and drilling mud. This optimized method offers a cost-effective and efficient path to high-quality nano-bentonite, solving the limitations of current purification methods.
研究领域:
材料科学
·
环境科学
·
化学工程
✓ 已验证
产品关联
DOI
高影响因子
CARBOHYDRATE POLYMERS
·
2020
·
IF 7.182
Yingying Cao,Yongqiang Zhang,Ying Zhang,Ling Wang,Lunchao Lv,Xifei Ma,Shaojuan Zeng,Hui Wang
Artemisinin is mainly derived from Artemisia annua L. Since the leaves composition is complex, artemisinin purification faces great challenges. In this work, functional chitosan membranes were fabricated by a one-step hydrolysis method through grafting long-chain alkyl group on the surface of chitosan to increase its hydrophobicity. The as-prepared membranes were used to adsorb wax oil ( i.e. , the impurity components) in Artemisia annua L. and to avoid co-precipitation of wax oil along with artemisinin using the crystallization technique for purification. Octyl-trimethoxysilane modified chitosan membrane (FCM-C 8 ) showed excellent capability to intensify this purification process. The product purity could reach more than 98 % using one crystallization step under the optimal conditions, and in this case, adsorption capacity of FCM-C 8 for wax oil was 478.9 mg/g. In addition, the adsorption kinetics and mechanism of wax oil on FCM-C 8 were studied. The membrane can simultaneously adsorb multiple components in wax oil through interactions like electrostatic forces, hydrogen bondings.
研究领域:
生物工程
·
制药工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2021
·
IF 3.978
Mianxing Luo,Meng Zhang,Guo Chen,Jun Zhao,Hongwei Guo
Functional carrier capable of covalently binding proteins from crude lysate under mild condition has become increasingly important in bioprocess. In this paper, a magnetic microbead that can covalently immobilize specific proteins directly from crude protein solution was developed, and its application conditions and stability were evaluated. The SpyCatcher was firstly modified with N-terminal cysteine by genetic methods and then the cys-SpyCatcher was covalently immobilized on epoxy-modified magnetic beads by epoxy-thiol reaction. The reaction between the epoxy and the only terminal thiol group in cys-SpyCatcher ensured the same orientation of the cys-SpyCatcher on the particle surface, while the maximum binding capacity of cys-SpyCatcher reached 23.6 μg/mg under the condition of 250 mM Na 2 SO 4 , pH 8.5 at initial cys-SpyCatcher concentration of 3 mg/mL. The functional magnetic microsphere ( [email protected] ) showed covalent site-specific capture property for enhanced green fluorescent protein fused with SpyTag (SpyTag-EGFP). A maximum binding capacity of (25 μg SpyTag-EGFP)/(mg beads) was far more than that of the previous carrier with SpyCatcher. In addition, the [email protected] could specifically capture the SpyTagged protein directly from the cell lysate, and this carrier showed good stability at pH 4–9 and temperature of 30–70ºC. Therefore, as a universal covalent capture carrier, it is believed that the [email protected] is useful in one-step purification and site-specific immobilization of the SpyTagged protein in bioprocess or biomedical application.
研究领域:
生物技术
·
生物化学
·
蛋白质工程
✓ 已验证
产品关联
DOI
高影响因子
SEPARATION AND PURIFICATION TECHNOLOGY
·
2024
·
IF 8.1
Xue-Jie Zhang,Wen-Cui Li,Ya-Qi Ba,Gang Li,Guang-Ping Hao,An-Hui Lu
The uptake of hydrogen (H 2 ) from natural gas pipelines (CH 4 ) through membrane separation technology is an efficient avenue of obtaining H 2 resources. Carbon membranes have the potential to be an attractive option for energy-efficient gas separations in the next generation of membranes due to their precise molecular discrimination ability and easy scalability. Here, we report a carbon membrane composed of a mesoporous substrate, seed layer and sieving layer, achieving a one-step uptake with 55.3 × 10 -9 mol·m −2 ·s −1 Pa −1 permeance and a separation factor for H 2 /CH 4 of 359.1 by feeding hydrogen (20 vol%). As revealed by the kinetic diffusion experiment, H 2 is transported in mesoporous substrate via Knudsen flow, differing from the transition flow taken by CH 4 , thus H 2 diffusion rates exceed CH 4 by an order of magnitude, which substantially increased H 2 purity from 20 vol% to higher purity as gas mixtures passed through the substrate. Then, the H 2 passed through the seed layer and sieving layer, resulting in a final H 2 purity of ∼ 99 vol%, a sharp increase from the initial low concentration. This carbon membrane integrates three functions of capture, pre-separation and purification into one material and provides a new solution for the uptake of H 2 from CH 4 .
研究领域:
材料科学
·
能源工程
✓ 已验证
产品关联
DOI
高影响因子
CHINESE JOURNAL OF CHEMICAL ENGINEERING
·
2022
·
IF 3.898
Zhaofeng Zhang,Juanjuan Ding,Min Wu,Bochao Liu,Huiwen Song,Shengping You,Wei Qi,Rongxin Su,Zhimin He
γ-Aminobutyric acid (GABA), a natural non-protein amino acid, plays an irreplaceable role in regulating the life activities of organisms. Nowadays, the separation and purification of food-grade GABA from fermentation broth is still a great challenge. This research utilized monosodium glutamate as a substrate for the production of high-purity GABA via an integrated process incorporating fermentation, purification, and crystallization. Firstly, 147 g·L −1 GABA with a yield of 99.8% was achieved through fed-batch fermentation by Lactobacillus brevis CE701. Secondly, three integrated purification methods by ethanol precipitation were compared, and crude GABA with a purity of 89.85% was obtained by the optimized method. Thirdly, GABA crystals with a purity of 98.69% and a yield of 60% were further obtained through a designed crystallization process. Furthermore, the GABA industrial production process model was established by Superproper Designer V10 software, and material balance and economic analysis were carried out. Ethanol used in the process was recovered with a recovery of 98.79% through Aspen simulated extractive distillation. Then the fixed investment (equipment purchase and installation costs) for an annual production of 80 t GABA will be about 833000 USD; the total annual production cost (raw material cost and utility cost) will be about 641000 USD. The annual sale of GABA may be at the range of 2400000–4000000 USD and the payback period will be about 1–2 year. This integrated process provides a potential way for the industrial-scale production of food-grade GABA.
研究领域:
生物工程
·
食品科学
✓ 已验证
DOI
高影响因子
JOURNAL OF RADIOANALYTICAL AND NUCLEAR CHEMISTRY
·
2022
·Vol. 331
·期 9
·3905-3913
·
IF 1.754
Guo Zong,Chu Taiwei
In this work, a separation and purification process of U(VI), Zr(IV) and I 2 from various fission products in nitric acid solution by 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide ([C 4 mim][NTf 2 ]) and dibutyl-3-(3-methylimidazolium)propylphosphoramide bis(trifluoromethanesulfonyl)imide-task specific ionic liquid (phosphoramide-TSIL) was investigated. The phosphoramide-TSIL was found to reduce ligand loss compared to dibutyl propylphosphoramidate by total organic carbon testing (TOC). The stripping of U(VI), Zr(IV) and I 2 could be achieved by guanidine carbonate solution, oxalic acid solution and sodium hydroxide solution, respectively. The phosphoramide-TSIL/[C 4 mim][NTf 2 ] showed good irradiation stability and recyclability. Phosphoramide-TSIL has the potential to be applied for separation and purification in the actual radioactive environment. Graphical abstract
研究领域:
核化学与放射化学
·
环境科学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF CHEMICAL RESEARCH
·
2023
·
IF 1.4
Ruilong Jia,Qizhuozhuo Hua,Xiaonan Ma,Guangfu Liao
Allyl alcohol and water can form the lowest boiling point azeotrope, but it is very hard to acquire high-purity allyl alcohol by normal distillation methods. Herein, a separation and purification protocol is developed for perfectly separating an azeotropic mixture of allyl alcohol and water using the salting-out method, in which three potassium phosphate salts, K5P3O10, K3PO4, and K4P2O7, are systematically investigated as salting-out agents, and finally a product consisting of > 99% allyl alcohol is obtained. A thermodynamic study demonstrates that this process involves endothermy and increment entropy. There is a good correlation between the solubility of allyl alcohol and the molar concentration of salt (mol per 1 kg water), and the solubility of the allyl alcohol in the organic phase or the water phase can be forecast using the mass percentage of salts. This work provides a new methodology for the efficient separation of an azeotropic mixture of allyl alcohol and water.
研究领域:
化学工程
·
物理化学
✓ 已验证
产品关联
DOI
高影响因子
MOLECULES
·
2021
·Vol. 26
·期 8
·2322
·
IF 4.412
Yunchang Fan,Di Wu,Sheli Zhang
Baicalin which has multiple biological activities is the main active component of the root ofScutellaria baicalensisGeorgi (SBG). Although its isolation and purification by adsorption methods have aroused much interest of the scientific community, it suffered from the poor selectivity of the adsorbents. In this work, an environmentally benign method was developed to prepare ionic liquids (ILs) grafted silica by using IL 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([C4mim]NTf2) and ethanol as reaction media. The IL 1-propyl-3-methylimidazolium chloride ([C3mim]Cl) grafted silica ([C3mim]+Cl−@SiO2) was used to adsorb and purify baicalin from the root extract ofScutellaria baicalensisGeorgi (SBG). Experimental results indicated that the adsorption equilibrium can be quickly achieved (within 10 min). The adsorption behavior of [C3mim]+Cl−@SiO2for baicalin was in good agreement with Langmuir and Freundlich models and the adsorption was a physisorption process as suggested by Dubinin–Radushkevich model. Compared with commercial resins, [C3mim]+Cl−@SiO2showed the strongest adsorption ability and highest selectivity. After desorption and crystallization, a purity of baicalin as high as 96.5% could be obtained. These results indicated that the ILs grafted silica materials were promising adsorbents for the adsorption and purification of baicalin and showed huge potential in the purification of other bioactive compounds from natural sources.Keywords:ionic liquids (ILs);grafted silica;adsorption;baicalin;purification
研究领域:
天然产物化学
·
植物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
·
2014
·Vol. 239
·期 5
·827-835
·
IF 1.387
Wang Panxue,Li Aixia,Dong Mei,Fan Mingtao
The aim of this study was to purify a malolactic enzyme (MLE) from Oenococcus oeni ( O. oeni ) strain and determine its properties in detail. O. oeni SD-2a was cultivated in the ATB broth supplemented with 7 g/L l -malic acid for harvesting the cells. After harvest, the cells were washed and disrupted for purification of MLE. MLE was purified from the supernatant of the disrupted cells through protamine sulfate precipitation, anion exchange chromatography and gel filtration chromatography. The purified MLE was identified using mass spectrometry. The MLE was purified by 43-fold with a yield of 0.42 % and possessed a specific activity of 419.2 U/mg. The purified enzyme with a nominal molecular mass of 59 kDa and a theoretical p I of 4.76 exhibited a maximum enzyme activity at 35 °C and pH 6.0, which retained over 50 % of its initial activity in the presence of 14 % (v/v) ethanol. Mn 2+ was proven to be the most effective divalent cation to promote enzyme activity. Under the conditions of temperature 30 °C and pH 6.0, the K
m and V
max of MLE on l -malic acid were 12.5 × 10 −3 M and 43.86 μmol/(min × mg), respectively. Moreover, the purified enzyme exhibited a higher stability with 0.1 M NaCl in addition and had a half-life of 30 days at 4 °C.
研究领域:
微生物学
·
酶学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）