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学者姓名：张燎原
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Abstract ：
To achieve the large-scale, low-cost preparation of acetaldehyde lyase (ALS), elastin-like polypeptides (ELPs) as non-chromatographic purification tags were employed to develop an ELP-ALS fusion protein in Escherichia coli. Induction expression results demonstrated that the ELPs tag efficiently improved the soluble expression of the ALS enzyme. Through two rounds of inverse transition cycling (ITC), highly pure ELP-ALS was obtained with an enzyme recovery rate of 85.77%, outperforming Ni2+-affinity chromatography (66.80%). The comparative analysis of enzymatic properties revealed that ELP fusion markedly improved the stability and substrate tolerance of the ALS enzyme. Kinetic parameter analysis under identical conditions showed that ELP-ALS possessed a Vmax of 15.25 U/mg and a kcat/Km of 73.05 s-1M-1, representing 1.86-fold and 2.97-fold improvements over His-ALS, respectively. Fed-batch reaction using ELP-ALS and acetaldehyde as biocatalyst and substrate, respectively, yielded 95.92 g/L acetoin with 49.32% increase compared to His-ALS (64.24 g/L). These results demonstrated the application potential of ELP-ALS as a promising biocatalyst for acetoin production from acetaldehyde due to its lower preparation cost, higher biocatalytic efficiency, better stability, and substrate tolerance.
Keyword ：
acetaldehyde lyase acetaldehyde lyase acetoin production acetoin production elastin-like polypeptides elastin-like polypeptides non-chromatographic purification non-chromatographic purification stability and substrate tolerance stability and substrate tolerance
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| GB/T 7714 | Lin, Hui , Zhang, Jiming , Hu, Jie et al. Improvement of Soluble Expression, Stability, and Activity of Acetaldehyde Lyase by Elastin-like Polypeptides Fusion for Acetoin Production from Acetaldehyde [J]. | BIOMOLECULES , 2025 , 15 (9) . |
| MLA | Lin, Hui et al. "Improvement of Soluble Expression, Stability, and Activity of Acetaldehyde Lyase by Elastin-like Polypeptides Fusion for Acetoin Production from Acetaldehyde" . | BIOMOLECULES 15 . 9 (2025) . |
| APA | Lin, Hui , Zhang, Jiming , Hu, Jie , Ma, Lu , Lai, Kaili , Zheng, Chaosong et al. Improvement of Soluble Expression, Stability, and Activity of Acetaldehyde Lyase by Elastin-like Polypeptides Fusion for Acetoin Production from Acetaldehyde . | BIOMOLECULES , 2025 , 15 (9) . |
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The development of simple, sensitive, and specific colorimetric methods for screening fentanyl (FTN) and its analogs is critically important for public health surveillance and drug control efforts. This study presents a novel multicolor colorimetric aptasensor based on gold nanobipyramid (AuNBP) growth for FTN and acetylfentanyl (AFTN) detection. First, a magnetic bead separation-assisted competitive binding system was developed using a high-affinity FTN aptamer (F27), which was demonstrated to specifically bind to FTN and AFTN, forming a parallel G-quadruplex structure. Then, we demonstrated that hydroquinone (HQ) can regulate AuNBP growth. This growth method is pH-compatible with a horseradish peroxidase (HRP) enzymatic reaction, leading to the development of a novel growth system termed HRP-catalyzed HQ-regulated AuNBP growth. By integrating these two systems, the developed aptasensor demonstrated similar signal responses and eight color changes for the detection of FTN and AFTN. The visual detection limit was 10 nM for both FTN and AFTN, and instrumental detection limits were 6.4 nM for FTN and 6.9 nM for AFTN. The aptasensor exhibited excellent sensitivity, specificity, and resistance to interference, and was successfully used to analyze spiked saliva and urine samples with a recovery of 95.9 %-102.9 % and an RSD (n = 5) < 5 %, making it a promising method for visual on-site detection of total FTN and AFTN concentration in human saliva and urine samples.
Keyword ：
Acetylfentanyl Acetylfentanyl Body fluids Body fluids Fentanyl Fentanyl Gold nanobipyramid growth Gold nanobipyramid growth Multicolor aptasensor Multicolor aptasensor Visual detection Visual detection
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| GB/T 7714 | Xu, Quanming , He, Xue , Luo, Guizhen et al. A sensitive multicolor colorimetric aptasensor for on-site detection of total fentanyl and acetylfentanyl concentration based on horseradish peroxidase-regulated gold nanobipyramid growth [J]. | MICROCHEMICAL JOURNAL , 2025 , 219 . |
| MLA | Xu, Quanming et al. "A sensitive multicolor colorimetric aptasensor for on-site detection of total fentanyl and acetylfentanyl concentration based on horseradish peroxidase-regulated gold nanobipyramid growth" . | MICROCHEMICAL JOURNAL 219 (2025) . |
| APA | Xu, Quanming , He, Xue , Luo, Guizhen , Liu, Jiayi , Wang, Zemiao , Zhu, Binling et al. A sensitive multicolor colorimetric aptasensor for on-site detection of total fentanyl and acetylfentanyl concentration based on horseradish peroxidase-regulated gold nanobipyramid growth . | MICROCHEMICAL JOURNAL , 2025 , 219 . |
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2,3-butanediol (2,3-BD) is a versatile bio-based platform chemical. An artificial four-enzyme synthetic biosystem composed of ethanol dehydrogenase, NADH oxidase, formolase and 2,3-butanediol dehydrogenase was designed for upgrading ethanol to 2,3-BD in our previous study. However, a key challenge in developing in vitro enzymatic systems for 2,3-BD synthesis is the relatively sluggish catalytic efficiency of formolase, which catalyzes the rate-limiting step in such systems. Herein, this study reports how engineering the tunnel and substrate binding pocket of FLS improved its catalytic performance. A series of single-point and combinatorial variants were successfully obtained which displayed both higher catalytic efficiency and better substrate tolerance than wild-type FLS. Subsequently, a cell-free biosystem based on the FLS:I28V/L482E enzyme was implemented for upgrading ethanol to 2,3-BD. Ultimately, this system achieved efficient production of 2,3-BD from ethanol by the fed-batch method, reaching a concentration of 1.39 M (124.83 g/L) of the product and providing both excellent productivity and yield values of 5.94 g/L/h and 92.7%, respectively. Taken together, this modified enzymatic catalysis system provides a highly promising alternative approach for sustainable and cost-competitive production of 2,3-BD.
Keyword ：
catalytic performance catalytic performance cell-free multienzyme catalysis cell-free multienzyme catalysis ethanol upgrading ethanol upgrading semi-rational design semi-rational design substrate tolerance substrate tolerance
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| GB/T 7714 | Zhang, Jiming , Lin, Hui , Zheng, Chaosong et al. Efficient 2,3-Butanediol Production from Ethanol by a Modified Four-Enzyme Synthetic Biosystem [J]. | MOLECULES , 2024 , 29 (16) . |
| MLA | Zhang, Jiming et al. "Efficient 2,3-Butanediol Production from Ethanol by a Modified Four-Enzyme Synthetic Biosystem" . | MOLECULES 29 . 16 (2024) . |
| APA | Zhang, Jiming , Lin, Hui , Zheng, Chaosong , Yang, Bin , Liang, Miao , Lin, Yi et al. Efficient 2,3-Butanediol Production from Ethanol by a Modified Four-Enzyme Synthetic Biosystem . | MOLECULES , 2024 , 29 (16) . |
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茯苓是一种我国传统药食同源常用的药用真菌,市场需求量极大,当前茯苓主要来源于以松木作为主料的椴木栽培,生物转化率低、菌林矛盾突出,而袋料茯苓栽培模式还不成熟,出苓产量与品质难以满足市场需求。根据茯苓培养过程中的酶系特点,筛选新基质原料,参考室外茯苓椴木栽培,开发茯苓设施化栽培工艺。利用该工艺室内栽培茯苓,7.5 kg袋料(56%含水量)平均可以采收2.3 kg鲜茯苓,鲜重转化率达54.8%,相对传统椴木栽培的20%生物转化率提高2.74倍;室内生产稳定,袋料茯苓大小均匀、形状圆整,商品性状良好,根据市场价格计算单袋产值23元,单袋成本11.9元,投入产出比达到193%,经济效益良好。对茯苓采收后的剩料进行开发利用,经小鼠喂养剩料实验发现大鼠血清的总抗氧化能力(TAS)和超氧化物歧化酶(T-SOD)活性提升显著;且可以作为益生元促进益生菌双歧杆菌和乳杆菌增殖效果,喂养后的小鼠肠道内乳杆菌和双歧杆菌数量显著提升,因此,剩料具有很大的应用开发前景。
Keyword ：
抗氧化 抗氧化 生物转化率 生物转化率 益生元 益生元 茯苓 茯苓 设施化栽培工艺 设施化栽培工艺
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| GB/T 7714 | 林辉 , 张燎原 . 茯苓设施化栽培工艺开发与应用 [C] //中国菌物学会2024年学术年会 . 2024 . |
| MLA | 林辉 et al. "茯苓设施化栽培工艺开发与应用" 中国菌物学会2024年学术年会 . (2024) . |
| APA | 林辉 , 张燎原 . 茯苓设施化栽培工艺开发与应用 中国菌物学会2024年学术年会 . (2024) . |
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Microbial xylanase has wide application in bioenergy, animal feed, environmental protection, the pulp and paper industry, and agricultural development. In this study, three xylanases from the microbiota of T. fuciformis with its companion strains were identified by metagenomics sequencing. The three enzymes were subjected to cloning and expression in E. coli or P. pastoris, purification, and characterization for their properties. The results showed that AsXyn1, from Annulohypoxylon stygium, among the three enzymes possessed high thermostability at 40 degrees C and broad pH tolerance in the range of 2.0-10.0, exhibiting its application potential. Furthermore, it was found that post-translational modification (such as glycosylation) of AsXyn1 enzyme modulated its activity, kinetic parameters, and thermostability. These results and findings provided a hint for enzyme modification and design in future.
Keyword ：
companion strain companion strain heterologous expression heterologous expression metagenome metagenome T. fuciformis T. fuciformis xylanase xylanase
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| GB/T 7714 | Lin, Yanhuan , Li, Changle , Wei, Chenxin et al. Mining, Identification, and Characterization of Three Xylanases from the Microbiota of T. fuciformis with Its Companion Strains [J]. | CATALYSTS , 2024 , 14 (1) . |
| MLA | Lin, Yanhuan et al. "Mining, Identification, and Characterization of Three Xylanases from the Microbiota of T. fuciformis with Its Companion Strains" . | CATALYSTS 14 . 1 (2024) . |
| APA | Lin, Yanhuan , Li, Changle , Wei, Chenxin , Lin, Hui , Zhang, Liaoyuan . Mining, Identification, and Characterization of Three Xylanases from the Microbiota of T. fuciformis with Its Companion Strains . | CATALYSTS , 2024 , 14 (1) . |
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On-site visual detection of organophosphorus pesticides (OPs) is crucial for safeguarding environmental and food safety. This study presents a robust, sensitive, and portable paper-based multicolor sensor (PMS) for the on-site detection of OPs. The PMS involves a novel Au+-induced gold nanobipyramids (AuNBPs) morphology transformation reaction within a paper-based analytical device (PAD) coupled with a bienzymatic hydrolysis system. The Au+-induced reaction sensitively regulates the aspect ratio of AuNBPs through Au+ disproportionation, which decreases the length of AuNBPs and simultaneously increases the width, accompanied by distinct multicolor changes. Upon performing in the PAD incorporating easily-prepared AuNBPs-deposited nylon membranes, a metal heater, and an oscillator, the Au+-induced reaction exhibits excellent reaction sensitivity, efficiency, reproducibility, AuNBPs stability, and color uniformity. In the bienzymatic hydrolysis system, acetylcholinesterase (AChE) and choline oxidase hydrolyze acetylcholine to product H2O2, which consumes tris(2-carboxyethyl)phosphine hydrochloride (TCEP), maintaining free Au+ for the disproportionation in the Au+-induced system, while the presence of OPs inhibits AChE activity, preserving TCEP to chelate Au+ and hindering the disproportionation. Under optimized conditions, the PMS exhibited eight dichlorvos concentration-corresponding color changes with a visual detection limit of 30 mu g/L and a spectrometry detection limit of 8.1 mu g/L. It achieved recoveries of 85.4-101.9 % in river water, drinking water, rice, and Chinese cabbage samples with relative standard deviations (n = 5) < 6 %. Thus, this study offers a portable, sensitive, reliable, and practical on-site detection method for OPs. Additionally, the proposed Au+-induced AuNBPs morphology transformation system within the PAD offers promising alternative strategies for developing other AuNBPs-based colorimetric sensors.
Keyword ：
Acetylcholinesterase Acetylcholinesterase Dichlorvos Dichlorvos Gold nanobipyramids Gold nanobipyramids Paper-based analytical device Paper-based analytical device Visual detection Visual detection
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| GB/T 7714 | Zhang, Feng , Ren, Enxi , Wang, Zemiao et al. Portable paper-based multicolor sensor for sensitive on-site detection of organophosphorus pesticides using a high-efficiency Au+-induced gold nanobipyramids aspect ratio regulation strategy [J]. | MICROCHEMICAL JOURNAL , 2024 , 207 . |
| MLA | Zhang, Feng et al. "Portable paper-based multicolor sensor for sensitive on-site detection of organophosphorus pesticides using a high-efficiency Au+-induced gold nanobipyramids aspect ratio regulation strategy" . | MICROCHEMICAL JOURNAL 207 (2024) . |
| APA | Zhang, Feng , Ren, Enxi , Wang, Zemiao , Fang, Ling , Yang, Weijuan , Zhang, Liaoyuan et al. Portable paper-based multicolor sensor for sensitive on-site detection of organophosphorus pesticides using a high-efficiency Au+-induced gold nanobipyramids aspect ratio regulation strategy . | MICROCHEMICAL JOURNAL , 2024 , 207 . |
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On-site quantitative detection of organophosphorus pesticides (OPs) is crucial for safeguarding food and public safety. This study presents a novel acetylcholinesterase (AChE)-mediated paper-based Au(3+)etching of gold nanobipyramids (AuNBPs) system. The system employs a long-term storable AuNBPs-deposited nylon membrane embedded within a portable and temperature-controlled paper-based analytical device. This system, coupled with a colorimeter-based quantitative method, enables the development of a practical paper-based multicolor sensor (PMS) for on-site quantitative detection of three common OPs (paraoxon, dichlorvos, and trichlorfon). In the absence of OPs, AChE hydrolyzes acetylthiocholine to thiocholine, which reduces Au(3+)to A(u+). The presence of OPs inhibits AChE activity, thereby preserving Au3+ to etch AuNBPs on nylon membranes, accompanied by multicolor changes. These color changes can be simply quantified by measuring the a* parameter of the CIELab color space using a portable colorimeter. Under optimal conditions, the PMS displayed eight OPs-corresponding color changes with a minimum detectable concentration of 1.0-10 mu g/L (visual observation) and limits of detection of 0.8-7.2 mu g/L (colorimeter) and 0.2-3.4 mu g/L (UV-vis spectrometry). The PMS successfully determined the OPs in vegetable and rice samples with recoveries of 89.0-109 % and RSDs (n = 5) of <6 %. These results were consistent with those obtained using the HPLC-MS method. The PMS demonstrates excellent portability, AuNBPs stability, detection sensitivity, and reproducibility, making it a promising tool for the on-site quantitative detection of OPs residues in food. Furthermore, the paper-based etching system coupled with the colorimeter-based quantitative method provides a valuable reference to develop practical PMSs for various targets in diverse fields.
Keyword ：
Colorimeter Colorimeter Colorimetric biosensor Colorimetric biosensor Gold nanobipyramids-deposited membrane Gold nanobipyramids-deposited membrane Paper-based analytical device Paper-based analytical device The CIELab color space The CIELab color space
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| GB/T 7714 | Zhang, Feng , Gao, Yu , Ren, Enxi et al. Paper-based multicolor sensor for on-site quantitative detection of organophosphate pesticides based on acetylcholinesterase-mediated paper-based Au3+-etching of gold nanobipyramids and CIELab color space [J]. | TALANTA , 2024 , 281 . |
| MLA | Zhang, Feng et al. "Paper-based multicolor sensor for on-site quantitative detection of organophosphate pesticides based on acetylcholinesterase-mediated paper-based Au3+-etching of gold nanobipyramids and CIELab color space" . | TALANTA 281 (2024) . |
| APA | Zhang, Feng , Gao, Yu , Ren, Enxi , Fang, Ling , Yang, Weijuan , Zhang, Liaoyuan et al. Paper-based multicolor sensor for on-site quantitative detection of organophosphate pesticides based on acetylcholinesterase-mediated paper-based Au3+-etching of gold nanobipyramids and CIELab color space . | TALANTA , 2024 , 281 . |
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采用已报道的灵芝磷酸葡萄糖变位酶(PGM)和UDP-葡萄糖焦磷酸化酶(UGPP)基因序列对茯苓基因组进行搜索比对，获得茯苓WcPGM和WcUGPP候选基因序列；以茯苓的cDNA为模板，成功克隆获得WcPGM和WcUGPP基因；随后利用pPIC9K构建2个基因的表达载体，并转化至毕赤酵母进行异源表达。序列分析表明WcPGM和WcUGPP基因全长分别为2 021 bp和2 144 bp,其中WcPGM基因含有5个外显子和4个内含子，编码564个氨基酸；WcUGPP基因含有11个外显子和10个内含子，编码502个氨基酸。氨基酸序列比对表明，WcPGM和WcUGPP与来源于绣球菌的ScPGM和ScUGPP的同源性分别为87%和91%。酶活测定表明，重组酶WcPGM可转化葡萄糖-1-磷酸(G1P)为葡萄糖-6-磷酸(G6P),酶活达1 540 U·mL~(-1);重组酶WcUGPP可催化G1P和尿苷三磷酸(UTP)合成UDP-葡萄糖(UDP-Glc),酶活达660 U·mL~(-1),表明从茯苓中筛选到的2个酶WcPGM和WcUGPP具有PGM和UGPP活性。分子模拟表明，WcPGM在催化可逆反应过程中S114为磷酸供体和受体，R24和K379作为催化酸和碱实现底物的磷酸化和去磷酸化，而E366和S368可实现底物的2种朝向，保障了中间产物葡萄糖-1,6-二磷酸的翻转，确保了WcPGM的可逆反应；而WcUGPP活性中心的核苷酸结合Loop和糖结合Loop保障了底物的正确朝向，K390作为催化碱实现了底物UTP/G1P与UDP-Glc的可逆反应。该研究为茯苓多糖合成途径的解析和代谢工程提升茯苓多糖产量奠定了基础。
Keyword ：
UDP-葡萄糖焦磷酸化酶 UDP-葡萄糖焦磷酸化酶 催化机制 催化机制 分子模拟 分子模拟 异源表达 异源表达 毕赤酵母 毕赤酵母 磷酸葡萄糖变位酶 磷酸葡萄糖变位酶 茯苓 茯苓
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| GB/T 7714 | 夏丽珍 , 李立志 , 张晓俊 et al. 茯苓多糖合成途径磷酸葡萄糖变位酶和UDP-葡萄糖焦磷酸化酶基因鉴定 [J]. | 化学与生物工程 , 2024 , 41 (06) : 44-52 . |
| MLA | 夏丽珍 et al. "茯苓多糖合成途径磷酸葡萄糖变位酶和UDP-葡萄糖焦磷酸化酶基因鉴定" . | 化学与生物工程 41 . 06 (2024) : 44-52 . |
| APA | 夏丽珍 , 李立志 , 张晓俊 , 张燎原 . 茯苓多糖合成途径磷酸葡萄糖变位酶和UDP-葡萄糖焦磷酸化酶基因鉴定 . | 化学与生物工程 , 2024 , 41 (06) , 44-52 . |
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本发明公开了一种具有热稳定性和底物耐受性乙醛裂合酶及应用，属于基因工程和酶工程技术领域。该酶是将乙醛裂合酶ALS‑I28V/L482E做单点突变或迭代突变，所得的酶具有更优异的热稳定性和催化效率，具有很好的工业应用价值。
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| GB/T 7714 | 张燎原 , 杨滢熙 , 胡捷 et al. 一种具有热稳定性和底物耐受性乙醛裂合酶及应用 : CN202410967264.8[P]. | 2024-07-18 . |
| MLA | 张燎原 et al. "一种具有热稳定性和底物耐受性乙醛裂合酶及应用" : CN202410967264.8. | 2024-07-18 . |
| APA | 张燎原 , 杨滢熙 , 胡捷 , 梁妙 . 一种具有热稳定性和底物耐受性乙醛裂合酶及应用 : CN202410967264.8. | 2024-07-18 . |
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2,3-丁二醇是一种典型的大宗化学品，广泛应用于化工、食品、医药、燃料等领域。生物法发酵制备2,3-丁二醇是当下的主要方法，然而，大多数报道的菌株为条件致病菌且发酵条件严格，限制了生物法制备2,3-丁二醇的发展。该研究以温泉底土壤为试验材料，经过分离、筛选与鉴定获得了1株高温条件下产2,3-丁二醇的甲基芽孢杆菌。通过摇瓶实验优化发酵工艺，确定最适发酵条件与最优培养基组成，利用优化后的发酵工艺甲基芽孢杆菌发酵24 h产物产量达到32.57 g/L。进一步进行上罐试验，基于不同发酵罐转速条件下甲基芽孢杆菌的产量积累情况，理性设计两阶段发酵策略，并以最优策略分批补料发酵制备2,3-丁二醇，2,3-丁二醇的产量在甲基芽孢杆菌发酵45 h时达到73.19 g/L,转化率达到90.25%。
Keyword ：
2 2 3-丁二醇 3-丁二醇 分批补料发酵 分批补料发酵 发酵优化 发酵优化 甲基芽孢杆菌 甲基芽孢杆菌 高温发酵 高温发酵
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| GB/T 7714 | 林燕环 , 陈飞雪 , 魏宸鑫 et al. 高温产2,3-丁二醇甲基芽孢杆菌的筛选及其发酵优化 [J]. | 食品与发酵工业 , 2024 , 50 (18) : 98-106 . |
| MLA | 林燕环 et al. "高温产2,3-丁二醇甲基芽孢杆菌的筛选及其发酵优化" . | 食品与发酵工业 50 . 18 (2024) : 98-106 . |
| APA | 林燕环 , 陈飞雪 , 魏宸鑫 , 李润志 , 张燎原 , 林辉 . 高温产2,3-丁二醇甲基芽孢杆菌的筛选及其发酵优化 . | 食品与发酵工业 , 2024 , 50 (18) , 98-106 . |
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