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学者姓名:王国增
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Biocatalytic technology has emerged as a powerful tool for synthesizing functional sugars with a wide range of physiological functions. D-lyxose isomerase is an important aldose-ketose isomerase used for synthesizing functional sugars such as L-ribose and D-mannose; however, at present, only a few D-lyxose isomerases have been studied. In this study, a metagenomic approach was employed to mine novel D-lyxose isomerases from the goat rumen microbiome. Eleven full-length D-lyxose isomerase genes were identified using sequence alignment and phylogenetic analysis. All 11 genes were classified as Group II D-lyxose isomerases and display low similarity to previously characterized D-lyxose isomerases. Five of the genes were selected for heterologous expression in Escherichia coli, and all were expressed successfully and had detectable enzyme activity. Further characterization revealed that optimum temperatures for the five enzymes range from 45 degrees C to 60 degrees C, and the optimum pH range is 7.0-8.0, with high relative residual activities under weakly acidic conditions. The recombinant enzymes have a broad substrate spectrum and are active against L-ribose, L-ribulose, D-fructose, and D-mannose, in addition to exhibiting the highest activity against D-lyxose. Among the five enzymes, GR-LI2 displays the highest activity toward L-ribulose, and GR-LI5 displays the highest activity against fructose, suggesting that these two enzymes have significant potential for L-ribose and D-mannose synthesis, respectively.
Keyword :
D-lyxose isomerase D-lyxose isomerase D-mannose D-mannose Functional sugar Functional sugar L -ribose L -ribose Metagenomics Metagenomics Rare sugar Rare sugar Rumen Rumen
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GB/T 7714 | Xu, Xin , Chen, Mingyi , Su, Yali et al. Identification and characterization of novel D-lyxose isomerases from the goat rumen metagenome [J]. | PROCESS BIOCHEMISTRY , 2025 , 149 : 213-221 . |
MLA | Xu, Xin et al. "Identification and characterization of novel D-lyxose isomerases from the goat rumen metagenome" . | PROCESS BIOCHEMISTRY 149 (2025) : 213-221 . |
APA | Xu, Xin , Chen, Mingyi , Su, Yali , Ding, Jiayun , Xie, Xiuye , Zhang, Xinyi et al. Identification and characterization of novel D-lyxose isomerases from the goat rumen metagenome . | PROCESS BIOCHEMISTRY , 2025 , 149 , 213-221 . |
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金属硫蛋白(metallothioneins,MTs)是一类低分子量、富含半胱氨酸残基且广泛分布于各类生物体的非酶金属结合蛋白,具有调控金属稳态、抵御氧化应激、屏蔽重金属等多种生物学功能,在食品、医药、环境、化妆品等领域具有广泛应用.关于MTs的基因发现、表达调控、结构表征、功能鉴定以及应用范围拓展等方面已有大量研究,但因其种类、结构和功能的多样性和复杂性,导致MTs还远未被完全了解.通过对全球范围内金属硫蛋白的相关研究文献进行分析,综述了金属硫蛋白领域的研究现状和动态热点,主要包括以下五个方面:金属硫蛋白的种类和结构;基因表达及其调控;锌稳态和铜稳态的调控等重要生物学功能;在生物医学领域中的角色和作用;在重金属污染生物治理领域的应用.未来金属硫蛋白研究趋向于:微生物源金属硫蛋白的开发;金属硫蛋白的高效表达及制备;建立或发展新的高分辨率的金属硫蛋白表征方法;探讨金属生物动力学与金属硫蛋白诱导表达之间的相关性以及金属硫蛋白的应用拓展.
Keyword :
生物学功能 生物学功能 生物标志物 生物标志物 种类结构 种类结构 表达调控 表达调控 金属硫蛋白 金属硫蛋白
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GB/T 7714 | 肖庆 , 沈艺鹏 , 王国增 . 金属硫蛋白研究现状及发展趋势 [J]. | 中国生物工程杂志 , 2025 , 45 (1) : 49-61 . |
MLA | 肖庆 et al. "金属硫蛋白研究现状及发展趋势" . | 中国生物工程杂志 45 . 1 (2025) : 49-61 . |
APA | 肖庆 , 沈艺鹏 , 王国增 . 金属硫蛋白研究现状及发展趋势 . | 中国生物工程杂志 , 2025 , 45 (1) , 49-61 . |
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Pseudostellaria heterophylla, also called Tai-zi-shen (TZS) in Traditional Chinese Medicine (TCM), is always used clinically to treat spleen deficiency symptoms. Polysaccharides in TZS have various pharmacological activities, including anti-diabetic, immune regulation, and myocardial protection. However, the relationship between the spleen-invigorating effects of TZS or its polysaccharides and intestinal flora are not clear. This study investigated the effects of TZS decoction (PHD) and polysaccharide (PHP) on immune function and intestinal flora in a rat model of spleen deficiency syndrome (SDS) induced by a decoction of raw rhubarb (RRD). PHD and PHP increased immune organ index, alleviated inflammatory cell filtration, and reduced the levels of pro-inflammatory cytokines in rats with spleen deficiency syndrome. In addition, the production of butyric acid was promoted in PHD and PHP groups. Moreover, 16S rRNA gene sequencing showed that PHD and PHP reduced the relative abundance of Firmicutes while increasing the one of Bacteroidetes; significantly increased the abundance of Lactobacillus and decreased the abundance of Rombutsia; and PHP significantly increased the abundance of Alloprevotella. And there was a significant positive correlation between the alleviation of SDS and short-chain fatty acids (SCFAs)-producing bacteria. These findings suggested PHD and PHP, especially PHP, has a potential to relieve spleen deficiency by reducing intestinal inflammation, modulating structure and composition of gut microbiota, and promoting the production of butyric acid.
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GB/T 7714 | Xiao, Qing , Zhao, Li , Jiang, Chang et al. Polysaccharides from Pseudostellaria heterophylla modulate gut microbiota and alleviate syndrome of spleen deficiency in rats [J]. | SCIENTIFIC REPORTS , 2022 , 12 (1) . |
MLA | Xiao, Qing et al. "Polysaccharides from Pseudostellaria heterophylla modulate gut microbiota and alleviate syndrome of spleen deficiency in rats" . | SCIENTIFIC REPORTS 12 . 1 (2022) . |
APA | Xiao, Qing , Zhao, Li , Jiang, Chang , Zhu, Yanjin , Zhang, Jizhou , Hu, Juan et al. Polysaccharides from Pseudostellaria heterophylla modulate gut microbiota and alleviate syndrome of spleen deficiency in rats . | SCIENTIFIC REPORTS , 2022 , 12 (1) . |
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Omega-transaminases (omega-TAs) have attracted considerable interest for their use in asymmetric synthesis of chiral amines with a high degree of optical purity and yield. The rapid evaluation of the characteristics of newly identified or engineered omega-TAs is important for industrial applications. In this study, a visible spectrophotometric assay was developed for rapid quantitative determination of omega-TA activities based on the transamination of 2-(4-nitrophenyl)ethan-1-amine to generate a red product (E)-N-(4-nitrophenethyl)-2-(4-nitrophenyl)ethen-1-amine. After various co-solvents were evaluated, dimethyl sulfoxide (DMSO) was considered optimal because the red product exhibits good solubility and retains its original color. The red product dissolved in DMSO has its highest absorbance at 465 nm, and its concentration has a good linear relationship with the absorbance. A spectro-photometric assay was established and validated using conventional HPLC analysis (<10% divergence). This method was then used to characterize an omega-TA from thermophilic Geobacillus thermoleovorans and an omega-TA ob-tained from the metagenome of a soda lake. The results demonstrated that omega-transaminase enzymatic properties could be characterized simply, rapidly, and at low cost, using this newly established visible spectrophotometric assay method.
Keyword :
Enzyme characterization Enzyme characterization Spectrophotometric assay Spectrophotometric assay ?-Transaminase ?-Transaminase UV UV Vis Vis
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GB/T 7714 | Wang, Guozeng , Jiang, Zhihao , Xiao, Qing et al. Visible spectrophotometric assay for characterization of omega-transaminases [J]. | ANALYTICAL BIOCHEMISTRY , 2022 , 658 . |
MLA | Wang, Guozeng et al. "Visible spectrophotometric assay for characterization of omega-transaminases" . | ANALYTICAL BIOCHEMISTRY 658 (2022) . |
APA | Wang, Guozeng , Jiang, Zhihao , Xiao, Qing , Jiang, Chang , Shi, Xian'ai . Visible spectrophotometric assay for characterization of omega-transaminases . | ANALYTICAL BIOCHEMISTRY , 2022 , 658 . |
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Proteinase K (PROK) from Parengyodontium album hydrolyzes keratin, a major protein component of poultry feathers, which are an inexpensive and renewable protein resource. Based on structural studies for analysis of amino acid flexibility near the catalytic center, identification of highly conserved residues, and experimental screening, we obtained a mutant R218S with residual activity 1.6-fold higher than that of PROK after incubation at 60 degrees C for 1 h. Molecular dynamics simulation indicated that substitution of Arg218 with Ser leads to three hydrogen bonds being introduced into the structure, stabilizing the beta-sheet in which Ser218 is located, and thus improvement of thermostability. Additionally, the mutant R218S had a 15% increase in specific activity compared to PROK and improvement in the rate and thoroughness of feather degradation compared with PROK. We confirmed the positive effects of enhancing catalytic center rigidity on enzyme thermostability, a finding which may have broad applications. (C) 2019 Elsevier B.V. All rights reserved.
Keyword :
B-factor B-factor Feather degradation Feather degradation Molecular modification Molecular modification Proteinase K Proteinase K Thermostability Thermostability
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GB/T 7714 | Ren, Yaxin , Luo, Huiying , Huang, Huoqing et al. Improving the catalytic performance of Proteinase K from Parengyodontium album for use in feather degradation [J]. | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2020 , 154 : 1586-1595 . |
MLA | Ren, Yaxin et al. "Improving the catalytic performance of Proteinase K from Parengyodontium album for use in feather degradation" . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 154 (2020) : 1586-1595 . |
APA | Ren, Yaxin , Luo, Huiying , Huang, Huoqing , Hakulinen, Nina , Wang, Yaru , Wang, Yuan et al. Improving the catalytic performance of Proteinase K from Parengyodontium album for use in feather degradation . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2020 , 154 , 1586-1595 . |
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碱性蛋白酶作为广泛应用的蛋白酶之一,具有重要的工业应用价值。发掘优质的碱性蛋白酶基因,实现其高效表达,有助于更好的满足工业应用需求。分别将Cordyceps fumosorosea和Beauveria bassiana来源的碱性蛋白酶基因pa1及pa2与表达载体pPIC9连接,并在毕赤酵母GS115中实现高效表达。对于纯化后的重组蛋白PA1及PA2的酶学性质进行测定,二者最适pH均为8.5,最适温度均为60℃,与同类酶相比具有一定的优势;PA1及PA2的温度稳定性较好,50℃下酶活保持稳定,60℃下处理10 min,二者均保持70%左右的酶活;PA1及PA2的pH耐受范围宽泛,在pH 4.0~...
Keyword :
克隆表达 克隆表达 毕赤酵母 毕赤酵母 碱性蛋白酶 碱性蛋白酶 酶学性质 酶学性质
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GB/T 7714 | 任雅馨 , 罗会颖 , 姚斌 et al. Cordyceps fumosorosea和Beauveria bassiana来源的碱性蛋白酶在毕赤酵母中的异源表达与性质测定 [J]. | 中国农业科技导报 , 2020 , 22 (11) : 69-78 . |
MLA | 任雅馨 et al. "Cordyceps fumosorosea和Beauveria bassiana来源的碱性蛋白酶在毕赤酵母中的异源表达与性质测定" . | 中国农业科技导报 22 . 11 (2020) : 69-78 . |
APA | 任雅馨 , 罗会颖 , 姚斌 , 王国增 , 涂涛 . Cordyceps fumosorosea和Beauveria bassiana来源的碱性蛋白酶在毕赤酵母中的异源表达与性质测定 . | 中国农业科技导报 , 2020 , 22 (11) , 69-78 . |
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Many methods have been used to detect heavy metals in herbal medicines, while few are developed to remove them. In this study, a novel genetically engineered fusion protein composed of metallothionein (MT), cellulose binding module (CBM), and superfolder GFP (sfGFP) was designed to remove heavy metals. MT, a kind of cysteine-rich protein, was used to chelate heavy metals with high specific affinity. The CBM facilitated the fusion protein MT-CBM-sfGFP binding to cellulose specifically, which made the purification and immobilization in one step. The sfGFP was used to detect the fusion protein MT-CBM-sfGFP easily during the process of expression and immobilization. The MT from Cancer pagurus (MTCap) and the CBM from Cellulomonas fimi (CBMcef) were used as an example and the fusion protein (MTCap-CBMcef-sfGFP) was expressed in Escherichia coli. Then, the cell lysates were mechanically mixed with cellulose to create biosorbent MTCap-CBMcef-sfGFP@cellulose. The efficiency of the biosorbent MTCap-CBMcef-sfGFP@cellulose for Pb2+ removal was evaluated using the water decoction of Honeysuckle as a model. Results suggested that MTCap-CBMcef-sfGFP@cellulose had high efficiency for Pb2+ removal from the water decoction of Honeysuckle without affecting its active ingredients. The low-cost, easy production, and high efficiency of the biosorbent enable it to have many applications in heavy metal removal from aqueous solutions of herbal medicines and food.
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GB/T 7714 | Xiao, Qing , Han, Jing , Jiang, Chang et al. Novel Fusion Protein Consisting of Metallothionein, Cellulose Binding Module, and Superfolder GFP for Lead Removal from the Water Decoction of Traditional Chinese Medicine [J]. | ACS OMEGA , 2020 , 5 (6) : 2893-2898 . |
MLA | Xiao, Qing et al. "Novel Fusion Protein Consisting of Metallothionein, Cellulose Binding Module, and Superfolder GFP for Lead Removal from the Water Decoction of Traditional Chinese Medicine" . | ACS OMEGA 5 . 6 (2020) : 2893-2898 . |
APA | Xiao, Qing , Han, Jing , Jiang, Chang , Luo, Meng , Zhang, Qingyi , He, Zhaodong et al. Novel Fusion Protein Consisting of Metallothionein, Cellulose Binding Module, and Superfolder GFP for Lead Removal from the Water Decoction of Traditional Chinese Medicine . | ACS OMEGA , 2020 , 5 (6) , 2893-2898 . |
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A new pullulanase-encoding gene, pLdSL3, was cloned from Alkalibacterium sp. SL3 and expressed in Escherichia coil. Deduced PulSL3 contains 2026 amino acid residues and consists of a putative signal peptide, four carbohydrate-binding modules, an E_set pullulanase domain, an alpha-amylase domain, and a pullulanase domain. The purified rPulSL3 exhibited pullulanase activity only. To identify the functions of the alpha-amylase and pullulanase domains, the N- and C-terminus truncated mutants (pulSL3 Delta N and pulSL3 Delta C) were also expressed in E. coli. The purified rPulSL3 Delta N had pullulanase activity, while rPulSL3 Delta C had no either alpha-amylase or pullulanase activity. Both rPulSL3 and rPulSL3 Delta N exhibited maximum activities at pH 9.0 and 50 degrees C, and were highly active and stable at the concentration of NaCl up to 4 M and resistant to a few surfactants and detergents. In comparison to the wild type, rPulSL3 Delta N had better thermostability (retaining 62.9 vs. 33.6% activity at 52 degrees C for 30 min), higher substrate affinity (with K-m of 0.10 vs. 0.15 mg mL(-1)) and higher catalytic efficiency (836.6 vs. 520.3 mL mg(-1) s(-1)). These excellent properties make rPulSL3 Delta N potential for the application in the industries of detergent, food, and environmental remediation.
Keyword :
Alkalibacterium Alkalibacterium Alkaline Alkaline Halotolerant Halotolerant Pullulanase Pullulanase SDS resistant SDS resistant Truncated mutant Truncated mutant
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GB/T 7714 | Huang, Han , Lin, Yun , Wang, Guozeng et al. Gene cloning, expression and biochemical characterization of a new multi-domain, halotolerant and SDS-resistant alkaline pullulanase from Alkalibacterium sp. SL3 [J]. | PROCESS BIOCHEMISTRY , 2020 , 96 : 1-10 . |
MLA | Huang, Han et al. "Gene cloning, expression and biochemical characterization of a new multi-domain, halotolerant and SDS-resistant alkaline pullulanase from Alkalibacterium sp. SL3" . | PROCESS BIOCHEMISTRY 96 (2020) : 1-10 . |
APA | Huang, Han , Lin, Yun , Wang, Guozeng , Lin, Juan . Gene cloning, expression and biochemical characterization of a new multi-domain, halotolerant and SDS-resistant alkaline pullulanase from Alkalibacterium sp. SL3 . | PROCESS BIOCHEMISTRY , 2020 , 96 , 1-10 . |
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传统的功能基因多样性研究主要采用PCR扩增片段构建文库及Sanger测序进行的,具有工作量大,耗时长,费用高和覆盖率低的不足。本研究采用基于Illumina HiSeq2500高通量测序平台对大布苏盐碱湖底泥宏基因组中的第10家族木聚糖酶基因多样性进行了研究。对扩增子测序后共获得227,420个读长,总长度约为49.5 Mbp。基于95%的一致性分析共获得467个可操作聚类单元(OTUs),其中有392个注释为木聚糖酶,比采用构建文库的方法获得的OTU数目要多300多个。这些OTU与GenBank数据库中的一致木聚糖酶蛋白序列的一致性范围为35-99%,其中75%与已知木聚糖酶序列的一致性在8...
Keyword :
基因多样性 基因多样性 宏基因组 宏基因组 木聚糖酶 木聚糖酶 盐碱湖 盐碱湖 高通量测序 高通量测序
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GB/T 7714 | 任雅馨 , 罗蒙 , 林娟 et al. 基于高通量扩增子测序的环境中木聚糖酶基因多样性分析 [C] //第十二届中国酶工程学术研讨会论文摘要集 . 2019 . |
MLA | 任雅馨 et al. "基于高通量扩增子测序的环境中木聚糖酶基因多样性分析" 第十二届中国酶工程学术研讨会论文摘要集 . (2019) . |
APA | 任雅馨 , 罗蒙 , 林娟 , 王国增 . 基于高通量扩增子测序的环境中木聚糖酶基因多样性分析 第十二届中国酶工程学术研讨会论文摘要集 . (2019) . |
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以大布苏盐碱湖分离菌Alkalibacterium sp.SL3的DNA为模板,利用PCR扩增 α-半乳糖苷酶基因(galSL3),构建重组质粒pET-22b-galSL3,转化至大肠杆菌BL21(DE3)诱导表达重组酶(rGalSL3).通过镍柱亲和层析分离纯化重组酶,并对纯化的重组酶进行性质研究.研究表明,纯酶rGalSL3最适pH值为5.5,最适温度为55℃;该酶在pH值5.0~10.0保持90%以上的剩余酶活,在50℃有非常好的热稳定性.在0~1.5 mol?L-1 NaCl溶液中该酶的酶活基本不受影响,在3.0 mol?L-1 NaCl溶液中有很好的稳定性.Pb2+、Ca2+、Co2+、Li+、Na+、K+、Triton-100和β-mercaptoethanol等对重组酶的活性有明显的促进作用.该酶水解对硝基苯基-α-D-吡喃葡萄糖苷的Km、vmax和kcat分别为(2.64±0.02)μmol?mL-1、(454.55±0.59)μmol?(mg?min)-1和(347.73±1.27)s-1.重组酶可水解蜜二糖和棉籽糖,不能水解瓜尔豆胶.
Keyword :
α-半乳糖苷酶 α-半乳糖苷酶 嗜盐嗜碱菌 嗜盐嗜碱菌 基因克隆 基因克隆 性质分析 性质分析 表达 表达
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GB/T 7714 | 邱聪花 , 罗蒙 , 林娟 et al. Alkalibacterium sp.SL3中α-半乳糖苷酶基因的克隆、 表达和性质研究 [J]. | 福州大学学报(自然科学版) , 2019 , 47 (1) : 136-142 . |
MLA | 邱聪花 et al. "Alkalibacterium sp.SL3中α-半乳糖苷酶基因的克隆、 表达和性质研究" . | 福州大学学报(自然科学版) 47 . 1 (2019) : 136-142 . |
APA | 邱聪花 , 罗蒙 , 林娟 , 叶秀云 , 王国增 . Alkalibacterium sp.SL3中α-半乳糖苷酶基因的克隆、 表达和性质研究 . | 福州大学学报(自然科学版) , 2019 , 47 (1) , 136-142 . |
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