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Oligo(dT)亲和层析介质的载量比较和机制分析

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高校化学工程学报 2024年第2期38卷 243-252页

作者：谭远志张鹏程孙艳娜张其磊姚善泾林东强浙江大学生物质化工教育部重点实验室浙江大学化学工程与生物工程学院浙江杭州310058

针对Oligo(d T)亲和层析介质的吸附性能,以poly(A)为模型分子,考察了4种Oligo(d T)亲和层析介质的静态吸附平衡、吸附动力学和动态结合载量(DBC),探讨了载量影响相关机制。结果表明,4种介质的合适吸附条件均为0.6 mol·L-1Na Cl、p H=6~... 详细信息

针对Oligo(d T)亲和层析介质的吸附性能,以poly(A)为模型分子,考察了4种Oligo(d T)亲和层析介质的静态吸附平衡、吸附动力学和动态结合载量(DBC),探讨了载量影响相关机制。结果表明,4种介质的合适吸附条件均为0.6 mol·L-1Na Cl、p H=6~7;Monomix d T20静态吸附容量最大,且poly(A)能扩散至介质微球深层孔内,而Poros Oligo(d T)25、Praesto Jetted (d T)25和Nano Gel d T20等3种介质中poly(A)均主要为表层吸附、静态吸附容量稍低;对于DBC,Nano Gel d T20和Monomix d T20的10%穿透的DBC较高,而Poros Oligo (d T)25和Praesto Jetted (d T)25相对略低。经分析,影响载量的主要因素包含基质种类、微球孔径、配基密度、间隔臂和配基长度等。对于基质种类,聚苯乙烯基质可能孔道结构较为特别。对于微球孔径,应针对不同大小的m RNA分子定制不同孔径的微球,以平衡传质阻力与可及吸附表面积之间的矛盾,从而增大DBC。

关键词：亲和层析 m RNA 寡脱氧胸腺苷酸载量生物分离

Removal of dyes from wastewater by growing fungal pellets in a semi-continuous mode

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Frontiers of Chemical Science and Engineering 2017年第3期11卷 338-345页

作者： Tao Lu Qilei Zhang shanjing yao Key Laboratory of Biomass Chemical Engineering of Ministry of Education College of Chemical and Biological Engineering Zhejiang University Hangzhou 310027 China

To increase the efficiency of dye removal from wastewater using mycelial pellets, a bubble column reactor with a simple structure was designed and efficiently used to remove dyes from solution containing dyes. The mycelial pellets were prepared by marine fungus Aspergillus niger ZJUBE-1. Eight dyes were tested as dye targets for the adsorption capacity of mycelial pellets and good removal results were obtained. Eriochrome black T was selected as a model dye for characterizing the adsorption processes in detail. The measurement results of Zeta potential and FT- IR analysis indicate that the electrostatic attraction may play a key role in the biosorption process. The bubble column reactor was utilized to study the batch dye-removal efficiency of mycelial pellets. A re-culture process between every two batches, which was under non-sterile condition, successfully enhanced the utilization of mycelium bio- mass. The dye removal rate is 96.4% after 12 h in the first batch and then decreases slowly in the following batches. This semi-continuous mode, which consists of commutative processes of dye-removal and re-culture, has some outstanding advantages, such as low power consumption, easy operation, high dye removal rate, and efficient biomass utilization.

关键词： dye mycelial pellets marine fungus bubble column reactor semi-continuous biosorption

Bi-/multi-modal pore formation of PLGA/hydroxyapatite composite scaffolds by heterogeneous nucleation in supercritical CO_2 foaming

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Chinese Journal of Chemical Engineering 2018年第1期26卷 207-212页

作者： Xin Xin Yixin Guan shanjing yao College of Chemical and Biological Engineering Zhejiang University

Scaffolds with multimodal pore structure are essential to cells differentiation and proliferation in bone tissue engineering. Bi-/multi-modal porous PLGA/hydroxyapatite composite scaffolds were prepared by supercritical C02 foaming in which hydroxyapatite acted as heterogeneous nucleation agent. Bimodal porous scaffolds were prepared under certain conditions, i.e. hydroxyapatite addition of 5%, depressurization rate of 0.3 MPa. min-1, soaking temperature of 55 ℃, and pressure of 9 MPa. And scaffolds presented specific structure of small pores （122 μM ± 66 μm） in the cellular walls of large pores （552 μm ±127 μm）. Furthermore, multimodal porous PLGA scaffolds with micro-pores （37 μM ± 11μM） were obtained at low soaking pressure of 7.5 MPa. The interconnected porosity of scaffolds ranged from （52.53 ± 2.69）% to （83.08±2.42）% by adjusting depressurization rate, while compression modulus satisfied the requirement of bone tissue engineering. Solvent-free CO2 foaming method is promising to fabricate bi-/multi-modal porous scaffolds in one step, and bioactive particles for osteogenesis could serve as nucleation agents.

关键词： Supercritical carbon dioxide Foam Tissue engineering PLGA scaffolds Hydroxyapatite Bi-/multi-modal pore

Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodolog

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Chinese Journal of Chemical Engineering 2013年第2期21卷 185-191页

作者： LUO Man GUAN Yixin yao shanjing Department of Chemical and Biological Engineering Zhejiang University Hangzhou 310027 China

Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was （377.9±1.7） mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.

关键词： disulfide bond formation protein A protein purification Box-Behnken experiment design response surface methodology multi-object programming

Efficient decolorization of dye-containing wastewater using mycelial pellets formed of marine-derived Aspergillus niger

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Chinese Journal of Chemical Engineering 2017年第3期25卷 330-337页

作者： Tao Lu Qilei Zhang shanjing yao Key Laboratory of Biomass Chemical Engineering of Ministry of Education College of Chemical and Biological EngineeringZhejiang UniversityHangzhou 310027China

In order to improve the efficient decolorization of dye-containing water by biosorbent and understand the biosorption mechanism, the self-immobilization mycelial pellets were prepared using a marine-derived fungus Aspergillus niger ZJUBE-1, and an azo dye, Congo red was chosen as a model dye to investigate batch decolorization efficiency by pellets. The pellets as biosorbent showed strong salt and acid tolerance in biosorption process. The results for dye adsorption showed that the biosorption process fitted well with models of pseudo-second-order kinetic and Langmuir isotherm, with a maximum adsorption capacity of 263.2 mg·g^(-1) mycelium. During 6 batches of continuous decolorization operation, the mycelial pellets could possess efficient decolorization abilities(>98.5%).The appearance of new peak in the UV–Vis spectral result indicated that the decolorization process may also contain biodegradation. The mechanism studies showed that efficient biosorption ability of pellets only relies on the active zone on the surface of the pellet, which can be enhanced by nutrition supplement or be shifted outward by a reculture process.

关键词： Mycelial pellets Batch decolorization Nutrition supplement Re-culture Dye wastewater treatment Adsorption mechanism

Repeated-batch Cultivation of Encapsulated Monascus purpureus by Polyelectrolyte Complex for Natural Pigment Production

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Chinese Journal of Chemical Engineering 2010年第6期18卷 1013-1017页

作者： LIU Jiefeng REN Yiran yao shanjing Department of Chemical and Biological Engineering Zhejiang University Hangzhou 310027 China Department of Biotechnology and Food Engineering Maoming College Maoming 525000 China

In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture.

关键词： microencapsulation Monascus purpureus natural pigment polyelectrolyte complex NaCS-PDMDAAC capsule cell immobilization

离子交换层析分离单抗电荷异质体的模型辅助过程优化

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化工学报 2024年第5期75卷 1903-1911页

作者：许茹枫陈煜成高丹焦静雨高栋王海彬姚善泾林东强浙江大学化学工程与生物工程学院生物质化工教育部重点实验室浙江杭州310058 海正生物制药有限公司浙江杭州311404

针对单抗电荷异质体分离,采用离子交换层析机理模型,预测洗脱分离行为,辅助工艺条件优化。设计了校准实验,拟合得到模型参数,模型计算与实验吻合良好,具有良好的预测能力。利用模型分析比较了不同洗脱方式,得到最优的两步阶跃洗脱方案,... 详细信息

针对单抗电荷异质体分离,采用离子交换层析机理模型,预测洗脱分离行为,辅助工艺条件优化。设计了校准实验,拟合得到模型参数,模型计算与实验吻合良好,具有良好的预测能力。利用模型分析比较了不同洗脱方式,得到最优的两步阶跃洗脱方案,具有较高的收率,但发现该分离过程对盐浓度极为敏感。进一步针对第一步洗脱盐浓度进行过程稳健性约束的过程优化,发现盐浓度为108.5 mmol/L时过程稳健性增强。经实验验证,两步阶跃洗脱收率最高可达到85.3%,稳健约束优化后第一步等度洗脱盐浓度操作区间增大为98.9~117.5 mmol/L。结果表明,模型辅助的工艺优化可以进行复杂条件分析,促进难分离体系的分离过程优化,并能够针对过程稳健性给出合理解决方案。

关键词：单克隆抗体电荷异质体分离离子交换层析层析模型过程优化

Adsorption behaviors of avian immunoglobulins and purification of immunoglobulin Y from chicken serum with mixed-mode resins

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Chinese Journal of Chemical Engineering 2019年第3期27卷 514-518页

作者： Yingdi Luo Qilei Zhang shanjing yao Dongqiang Lin Key Laboratory of Biomass Chemical Engineering of Ministry of Education College of Chemical and Biological Engineering Zhejiang University

The importance of immunoglobulin Y(IgY) as a specific antibody equivalent to mammalian immunoglobulin G(IgG) is well recognized. However, production of highly purified IgY is still difficult due to the lack of specific purification methods. In this study, adsorption behaviors of Ig Y on four mixed-mode resins with functional ligands of 4-mercatoethyl-pyridine(MEP), 2-mercapto-1-methyli-midazole(MMI), 5-aminobenzi-midazole(ABI) and tryptophan-5-aminobenzi-midazole(W-ABI) were evaluated. The results showed that high adsorption ratio were found at p H 6.0–7.0 with little adsorption under acidic conditions. The resin with ABI ligand was then used to separate IgY from immunized chicken serum. An efficient process with Ig Y purity of 95% and recovery of 90% was developed after optimization of loading and elution p H and injection volume. The biological activity of the purified Ig Y was fully maintained. These results indicated that mixed-mode chromatography with specially-designed ligands has great potential for the separation of Ig Y from crude feedstock.

关键词： Mixed-mode chromatography Immunoglobulin Y Adsorption selectivity Separation

Thermostable ethanol tolerant xylanase from a cold-adapted marine species Acinetobacter johnsonii

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Chinese Journal of Chemical Engineering 2019年第5期27卷 1166-1170页

作者： Dongsheng Xue Xuhao Zeng Dongqiang Lin shanjing yao Key Laboratory of Fermentation Engineering (Hubei Universty of Technology) Ministry of EducationHubei Provincial Cooperative Innovation Center of Industrial FermentationHubei Key Laboratory of Industrial MicrobiologyHubei University of TechnologyWuhan 430068China Department of Chemical and Bioengineering Zhejiang UniversityHangzhou 310027 China

A xylanase-producing bacterium, isolated from deep sea sediments, was identified as the cold-adapted marine species Acinetobacter Johnsonii. A cold-adapted marine species Acinetobacter Johnsonii could grow at 4 ℃. The optimum temperature and pH of xylanase from a cold-adapted marine species Acinetobacter Johnsonii were 55 ℃ and pH 6.0. Xylanase from a cold-adapted marine species Acinetobacter Johnsonii remained at 80% activity after incubation for 1 h at 65 ℃. The xylanase activity was 1.2-fold higher in 4% ethanol solution than in ethanol free solution. Gibbs free energy of denaturation, ΔG, was higher in 4% ethanol solution than in ethanol free solution. Thermostable ethanol tolerant xylanase was valuable for bioethanol production by simultaneous saccharification and fermentation process with xylan as a carbon source.

关键词： Xylanase Ethanol tolerant Thermostable Cold-adapted Acinetobacter Johnsonii