APPLIED SURFACE SCIENCE
·
2021
·
IF 6.707
Wenting He,Hao Shen,Zixin Zhou,Ze Huang,Hao Chao,Jiayi Song,Ping Su,Yi Yang
Metal-organic frameworks (MOFs) are excellent platforms for enzyme immobilization. However, strong or weak interactions between enzymes and MOFs often lead to inactivation and enzyme leaching. Herein, single-stranded DNA (ssDNA) with Janus properties was used as a linker between enzymes and the dual carriers of MOFs and graphene oxide (GO), which provided a simple and effective platform to tightly co-immobilize glucose oxidase (GO X ) and horseradish peroxidase (HRP). Mechanistic investigation revealed that the enzyme was immobilized mainly through π-π stacking between nucleobases and GO and coordination of the sugar-phosphate backbone with zirconium clusters in MOFs. This strategy ameliorated traditional surface adsorption issues for enzyme immobilization on MOFs. By introducing GO and ssDNA, the GO X &HRP cascade reaction on MOFs was enhanced significantly, showing better kinetics than the corresponding free GO X &HRP. Furthermore, the immobilized GO X &HRP exhibited substrate selectivity and a wide linear range for glucose detection. This enzyme immobilization strategy should have broad application prospects and practical value in biocatalysis and diagnostics.
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ANALYST
·
2021
·Vol. 146
·期 18
·5732-5739
·
IF 4.616
Tingting Zhao,Shiqian Tong,Susu Zhou,Chenchen Lin,Xucong Lin,Zenghong Xie
Developing a functional affinity monolithic column towards in-tube solid-phase microextraction (IT-SPME) for selective sample pretreatment is critical. Herein, a high-performance capillary affinity monolithic column with an ultra-high aptamer coverage density was rapidly fabricated via a simple adsorption strategy, in which aptamers with natural sequences were directly immobilized on an ammonium-based strongly cationic matrix. Limitations of the traditional biological or covalent methods such as time-consuming modification reactions, special requirement of active groups (e.g. –NH2 and –SH) on the aptamer, and low aptamer coverage density levels were avoided. An ultra-high coverage density of 8616 pmol μL−1 was achieved with excellent stability, and the highest aptamer-modification level among all the current methods was reached. Selective recognition and high recovery yields of the model mycotoxin ochratoxin A (OTA) were achieved in 95.9 ± 0.98%–97.9 ± 0.28% (n = 3). In particular, there was little cross-reactivity towards the OTB analogue of only 0.5% even in the case of 250 fold of the analogue OTB, which was not reported in previous affinity monoliths. Upon sample analysis, satisfactory discriminations of trace OTA were obtained at 93.7 ± 1.4%–95.5 ± 2.5% (n = 3) in beer and wheat. A facile and direct method for efficiently fabricating an aptamer-based affinity monolith towards online selective IT-SPME was proposed.
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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.
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TALANTA
·
2019
·
IF 4.916
Fangfang Chen,Mao Mao,Jiayu Wang,Jiawei Liu,Feng Li
One viable solution to improve the conformational stability of template proteins is to use multiple, weaker modes of action to immobilize proteins on the surfaces of a solid support. Herein, we introduce a novel surface imprinting technique for human serum albumin (HSA) by a dual immobilization/imprinting strategy. Specifically, HSA was first conjugated to the surfaces of magnetic Fe 3 O 4 nanoparticles through a reversible aldmine condensation reaction . Dopamine (DA) was then used to imprint the protein template via an auto-polymerization reaction in biocompatible aqueous media. The resultant magnetic molecular imprinted polymers (MMIPs) possess high adsorption capacity (70.2 mg g −1 ), superior selectivity ( IF = 4.54), and rapid capturing kinetics to HSA (within 20 min). We successfully demonstrate the practical applicability of MMIPs to the selective removal of HSA from human serum sample. Our work offers a novel and robust solution to develop proteins imprinted materials with high binding capacity and selectivity. We anticipate such materials will find wide applications to protein detection or removal in diverse real-life clinical and biological samples.
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APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2019
·Vol. 188
·期 3
·677-689
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IF 2.14
Zhao Jiong-feng,Lin Jian-ping,Yang Li-rong,Wu Mian-bin
Nano-sized Fe 3 O 4 was synthesized by chemical co-precipitation and subsequently modified with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde to introduce aldehyde group on its surface. With the help of “interface activation” by adding sucrose esters-11 as surfactant, lipase from Rhizopus oryzae was successfully immobilized onto the carrier with great enhancement of activity. The hydrolysis activity of immobilized enzyme were 9.16 times and 31.6 times of free enzyme when p-nitrophenol butyrate and p-nitrophenol palmitate were used as substrates. The thermo-stability of immobilized enzyme was also enhanced compared to free enzyme. The immobilized enzyme was successfully applied in synthesis of 1,3-diacyglycerols (1,3-DAG). The specific esterification activity of immobilized enzyme was about 1.5 times of the free enzyme. The immobilized enzyme showed good region-selectivity towards 1,3-diacyglycerols and retained nearly 80% of its activity after reused for 60 times, revealing a good industrial application prospect.
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ADVANCED FUNCTIONAL MATERIALS
·
2019
·Vol. 29
·期 17
·1807191
·
IF 15.621
Zequn Yang,Hailong Li,Jianping Yang,Shihao Feng,Xi Liu,Jiexia Zhao,Wenqi Qu,Pu Li,Yong Feng,Po-Heng Lee,Kaimin Shih
A key challenge in elemental mercury (Hg 0 ) decontamination from flue gas lies in the design of a sorbent with abundant reactive adsorption sites that exhibit high affinity toward Hg 0 to simultaneously achieve rapid capture and large capacity. Herein, zeolitic imidazolate framework-8 (ZIF-8) supported copper selenide (CuSe) nanocomposites are synthesized by a newly designed two-step surfactant-assisted method. The as-prepared CuSe/ZIF-8 with CuSe to ZIF-8 mass ratio of 80% (0.8NC-ZIF) exhibits unparalleled performance toward Hg 0 adsorption with equilibrium capacity and average rate reaching 309.8 mg g −1 and 105.3 µg g −1 min −1 , respectively, surpassing all reported metal sulfides and traditional activated-carbon-based sorbents. The impressive performance of 0.8NC-ZIF for Hg 0 immobilization is primarily attributed to the adequate exposure of the Se-terminated sites with high affinity toward Hg 0 resulted from the layered structure of CuSe. The adsorbed mercury selenide exhibits even higher stability than the most stable natural mercury ore—that is, mercury sulfide—hence minimizing its environmental impact when the CuSe/ZIF-8 sorbent is dumped. This work provides a new mindset for future design of sorbents for efficient Hg 0 capture from industrial flue gas. The results also justify the candidature of CuSe/ZIF to be applicable for mercury pollution remediation in real-world conditions.
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COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
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2019
·
IF 3.131
Nan Zhou,Yifan Wang,Liyang Huang,Jingang Yu,Huanli Chen,Jiajie Tang,Fengjuan Xu,Xiangyang Lu,Mei-e Zhong,Zhi Zhou
Surface modification of biochar is significant to obtain more functionalized material for contaminants immobilization. In situ modification, modifying the biochar while the conversion process, was applied by a novel wet pyrolysis system. By using phosphoric acid as working medium, liquid/air interface was created to introduce low content of oxygen, under the control of appropriately settled reaction temperature. Followed with the addition of sodium carbonate to neutralize the system and increase mineral species, effective biochar adsorbent attached with large quantity of acidic oxygenic and phosphoric groups, and carbonate anions was finally produced. The biochar showed excellent decontamination performance of lead with a high adsorption capacity of 316 mg·g −1 and removal efficiency larger than 95%. The alteration of the characteristics after the adsorption confirmed the adsorption mechanism is the bi-function of surface coordination and precipitation. This novel route is easy handling and energy saving, promising it to be a reliable way to produce biochars with attractive surface properties for environmental decontamination.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2019
·
IF 3.571
Si Lin,Zhen Qin,Qiming Chen,Liqiang Fan,Jiachun Zhou,Liming Zhao
Chitooligosaccharide has been reported to possess diverse bioactivities. The development of novel strategies for obtaining optimum degree of polymerization (DP) chitooligosaccharides has become increasingly important. In this study, two glycoside hydrolase family 46 chitosanases were studied for immobilization on curdlan (insoluble β-1,3-glucan) using a novel carbohydrate binding module (CBM) family 56 domain from a β-1,3-glucanase. The CBM56 domain provided a spontaneous and specific sorption of the fusion proteins onto a curdlan carrier, and two fusion enzymes showed increased enzyme stability in comparison with native enzymes. Furthermore, a continuous packed-bed reactor was constructed with chitosanase immobilized on a curdlan carrier to control the enzymatic hydrolysis of chitosan. Three chitooligosaccharide products with different molecular weights were prepared in optimized reaction conditions. This study provides a novel CBM tag for the stabilization and immobilization of enzymes. The controllable hydrolysis strategy offers potential for the industrial-scale preparation of chitooligosaccharides with different desired DPs.
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ENZYME AND MICROBIAL TECHNOLOGY
·
2018
·
IF 2.932
Juan Han,Yunfeng Cai,Yun Wang,Lei Gu,Chunmei Li,Yanli Mao,Wenli Zhang,Liang Ni
In this work, a novel Ni 2+ &AABOB bifunctional core–shell magnetic composite nanoparticles (MNPs), Fe 3 O 4 @p(AABOB-co-AIM-Ni 2+ ), were synthesized for the immobilization of horseradish peroxidase (HRP). Two kinds of interaction are simultaneously incorporated to improve the stability and reusability of immobilized HRP: one is the coordination between Ni 2+ and amide group in HRP, and the other is the interaction of the wulff-type boronic acid group with cis-diol-containing HRP which belongs to a type of glycoproteins. Accordingly, the proposed approach achieved high enrichment capacity (174.75 mg/g, protein/beads), and the activity of immobilized HRP still achieved 80% after reuse for 5 cycles. The degradation efficiency of phenol in the presence of immobilized HRP reached 93% within 10 min, which was significantly higher than that of free HRP only about 50% within 20 min. In addition, the immobilized HRP can be easily separated from the reaction solution by an external magnetic field.
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BIOCHEMICAL ENGINEERING JOURNAL
·
2018
·
IF 3.226
Amjad Hussain Memon,Runsheng Ding,Qipeng Yuan,Hao Liang,Yun Wei
Enhancing activity of multiple enzymes by co-localization are key topics in bionanotechnology. Herein, we introduce a facile co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP) in the matrix of Cu (copper) and GMP (guanosine 5′-monophosphate). The multienzyme system by co-immobilizing GOx&HRP observed with high catalytic efficiency and stability compared with free GOx&HRP enzymes. GOx&HRP@Cu/GMP stability was observed 40% increased at pH 3 and thermal stability more than 70% determined at 90 ᵒ C. The storage stability was sustained 8 times higher than free enzymes after thirteen days and reusability after reacting eight times with fresh substrate retains 90% relative activity. Further, GOx&HRP@Cu/GMP demonstrated an excellent selectivity in the development of glucose biosensor and exhibited good resistance against the protease enzymes and organic solvents. The obtained results emphasize that co-immobilization of GOx&HRP in the matrix of Cu/GMP could be an efficient protocol for the biocatalyst encapsulation.
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Energy & Environmental Science
·
2018
·Vol. 11
·期 9
·2620-2630
·
IF 30.067
Yingze Song,Wen Zhao,Long Kong,Li Zhang,Xingyu Zhu,Yuanlong Shao,Feng Ding,Qiang Zhang,Jingyu Sun,Zhongfan Liu
Lithium–sulfur (Li–S) batteries are deemed as one of the most promising next-generation energy storage systems. However, their practical application is hindered by existing drawbacks such as poor cycling life and low Coulombic efficiency due to the shuttle effect of lithium polysulfides (LiPSs). We herein present an in situ constructed VO2–VN binary host which combines the merits of ultrafast anchoring (VO2) with electronic conducting (VN) to accomplish smooth immobilization–diffusion–conversion of LiPSs. Such synchronous advantages have effectively alleviated the polysulfide shuttling, promoted the redox kinetics, and hence improved the electrochemical performance of Li–S batteries. As a result, the sulfur cathode based on the VO2–VN/graphene host exhibited an impressive rate capability with ∼1105 and 935 mA h g−1 at 1C and 2C, respectively, and maintained long-term cyclability with a low capacity decay of 0.06% per cycle within 800 cycles at 2C. More remarkably, favorable cyclic stability can be attained with a high sulfur loading (13.2 mg cm−2). Even at an elevated temperature (50 °C), the cathodes still delivered superior rate capacity. Our work emphasizes the importance of immobilization–diffusion–conversion of LiPSs toward the rational design of high-load and long-life Li–S batteries.
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Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 36
·5718-5728
·
IF 4.776
Jiayi Song,Hao Shen,Ye Yang,Zixin Zhou,Ping Su,Yi Yang
The overall stability and activity of immobilized enzyme systems have suffered from non-specific adsorption immobilization of enzymes at the surface of carriers through electrostatic and hydrophobic interactions. Elimination of these non-specific adsorptions of enzymes on the surface of carriers is critical for enzyme reactions. Herein, for the first time, we have prepared zwitterion-functionalized magnetic particles with amino, phosphonate, and thiol functional groups to coimmobilize glucose oxidase and horseradish peroxidase by DNA directed immobilization. The zwitterionic surface of the multifunctional magnetic particles could efficiently suppress non-specific adsorption of different kinds of enzymes, and the immobilized multienzyme catalyst without non-specifically adsorbed enzymes exhibited excellent enzymatic activity, stability, and reusability compared with those of free and non-specifically adsorbed enzymes. The immobilized multienzymes maintained 87% of their enzymatic activity after two weeks of storage at 4 °C and 58% of their enzymatic activity after 90 min of thermal incubation at 60 °C. Furthermore, the immobilized multienzymes exhibited more than 87% enzymatic activity after reuse for 10 cycles. The apparent Km and catalytic efficiency (kcat/Km) values of the immobilized multienzymes were 12.6 mM and 4.03 s−1 mM−1, respectively, which were 0.2- and 11.8-fold better than those of the free enzymes, indicating effective cascade efficiency and substrate affinity. The optimal ratio of GOx and HRP was 1 : 5, and the prepared immobilized enzymes could detect low concentrations of glucose (0.5 μM) with excellent selectivity. Therefore, we believe that the strategy developed in this study can be widely applied in biotechnology, industrial catalysis, and biomedical engineering.
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