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学者姓名：叶秀云
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Abstract ：
The white rot fungus Cerrena unicolor 87613 has been previously shown to be a promising resource in laccase production, an enzyme with significant biotechnological applications. Conventional methods face technical challenges in improving laccase activity. Attempts are still being made to develop novel approaches for further enhancing laccase activity. This study aimed to understand the regulation of laccase activity in C. unicolor 87613 for a better exploration of the novel approach. Transcriptomic and metabolomic analyses were performed to identify key genes and metabolites involved in extracellular laccase activity. The findings indicated a strong correlation between the glutathione metabolism pathway and laccase activity. Subsequently, experimental verifications were conducted by manipulating the pathway using chemical approaches. The additive reduced glutathione (GSH) dose-dependently repressed laccase activity, while the GSH inhibitors (APR-246) and reactive oxygen species (ROS) inducer (H2O2) enhanced laccase activity. Changes in GSH levels could determine the intracellular redox homeostasis in interaction with ROS and partially affect the expression level of laccase genes in C. unicolor 87613 in turn. In addition, GSH synthetase was found to mediate GSH abundance in a feedback loop. This study suggests that laccase activity is negatively influenced by GSH metabolism and provides a theoretical basis for a novel strategy to enhance laccase activity by reprogramming glutathione metabolism at a specific cultivation stage.
Keyword ：
Cerrena unicolor Cerrena unicolor glutathione metabolism glutathione metabolism laccase activity laccase activity metabolome metabolome transcriptome transcriptome
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| GB/T 7714 | Zhang, Long-Bin , Qiu, Ting-Ting , Qiu, Xiu-Gen et al. Transcriptomic and metabolomic analysis unveils a negative effect of glutathione metabolism on laccase activity in Cerrena unicolor 87613 [J]. | MICROBIOLOGY SPECTRUM , 2024 , 12 (2) . |
| MLA | Zhang, Long-Bin et al. "Transcriptomic and metabolomic analysis unveils a negative effect of glutathione metabolism on laccase activity in Cerrena unicolor 87613" . | MICROBIOLOGY SPECTRUM 12 . 2 (2024) . |
| APA | Zhang, Long-Bin , Qiu, Ting-Ting , Qiu, Xiu-Gen , Yang, Wu-Wei-Jie , Ye, Xiu-Yun , Meng, Chun . Transcriptomic and metabolomic analysis unveils a negative effect of glutathione metabolism on laccase activity in Cerrena unicolor 87613 . | MICROBIOLOGY SPECTRUM , 2024 , 12 (2) . |
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This study sought to understand how the features of proteins impact the properties of nanoparticles assembled using the pH-shifting approach and the mechanism behind. Four legume protein isolates from faba bean, mung bean, soy, and pea were fractionated into natural aqueous-soluble (Sup) and aqueous-insoluble (Sed) fractions, which were proved to serve as shell and core, respectively, for the pH-driven-assembled nanoparticles. Using zein instead of Sed fractions as the core improved size uniformity, and particle size can be precisely controlled by adjusting core/shell ratios. Using the proteomic technique and silico characterization, the features of identified proteins indicated that hydrophobicity rather than molecular weight, surface charge, etc., mainly determined particle size. With molecular docking, structural analysis, and dissociation tests, the assembly of zein/Sup-based nanoparticles was dominantly driven by hydrophobic interactions. This study provides constructive information on the correlation between protein features and the properties of pH-driven-assembled nanoparticles, achieving a precise control of particle size.
Keyword ：
assembly mechanism assembly mechanism legume proteins legume proteins LFQ proteomics LFQ proteomics pH-shifting nanoparticulation pH-shifting nanoparticulation zein zein
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| GB/T 7714 | Mao, Yuhong , Huang, Wenting , Jia, Rongju et al. Correlation between Protein Features and the Properties of pH-Driven-Assembled Nanoparticles: Control of Particle Size [J]. | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2023 , 71 (14) : 5686-5699 . |
| MLA | Mao, Yuhong et al. "Correlation between Protein Features and the Properties of pH-Driven-Assembled Nanoparticles: Control of Particle Size" . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 71 . 14 (2023) : 5686-5699 . |
| APA | Mao, Yuhong , Huang, Wenting , Jia, Rongju , Bian, Yangyang , Pan, Min-Hsiung , Ye, Xiuyun . Correlation between Protein Features and the Properties of pH-Driven-Assembled Nanoparticles: Control of Particle Size . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2023 , 71 (14) , 5686-5699 . |
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Phenolic compounds are widely distributed in nature and industrial environment, and their detoxifi-cation or bioactive enhancement is of great value to environmental protection and industrial develop-ment. Laccases are multicopper oxidases that catalyse the oligo-or polymerisation of phenolic compounds. Identifying new laccase producers and investigating their application potential are of great importance. In this study, a white-rot fungus, Trametes hirsuta EZ1, with significantly high laccase pro-ductivity was isolated. The optimum conditions were studied for the maximum fermentation of extra-cellular laccase, which was achieved at 150 U/mL with a medium containing 10% strain EZ1, 7% maltodextrin, 1.5% peptone, and 0.5 mM Cu2+, and incubation at initial pH 6.0, 32 degrees C, and 180 rpm for nine days. Subsequently, a 70-kDa laccase was purified that showed activity over a wide range of tem-perature and pH, sensitivity to many metal ions and sodium dodecyl sulphate, and high tolerance to organic solvents. Purified laccase showed a significant unreported effect by catalysing catechol or ferulic acid into dimers, trimers, and tetramers or caffeic acid into dimers, trimers, tetramers, and pentamers. The oligomeric mixtures exhibited increased antioxidative capacity compared to that of each parent monomer, except for caffeic acid derivatives. Our study offers a novel strain source for laccase production and broadens its application in the enhancement of bioactive compounds.(c) 2022 British Mycological Society. Published by Elsevier Ltd. All rights reserved.
Keyword ：
Caffeic acid Caffeic acid Catechol Catechol Ferulic acid Ferulic acid Laccase Laccase Oligomerisation Oligomerisation Trametes hirsuta Trametes hirsuta
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| GB/T 7714 | Zhang, Long-Bin , Deng, Zhi-Qiang , Qiu, Ting-Ting et al. Characterisation of a laccase isolated from Trametes hirsuta and its application in the oligomerisation of phenolic compounds [J]. | FUNGAL BIOLOGY , 2023 , 127 (1-2) : 872-880 . |
| MLA | Zhang, Long-Bin et al. "Characterisation of a laccase isolated from Trametes hirsuta and its application in the oligomerisation of phenolic compounds" . | FUNGAL BIOLOGY 127 . 1-2 (2023) : 872-880 . |
| APA | Zhang, Long-Bin , Deng, Zhi-Qiang , Qiu, Ting-Ting , Yang, Wu-Wei-Jie , Zhu, Fan , Ye, Xiu-Yun . Characterisation of a laccase isolated from Trametes hirsuta and its application in the oligomerisation of phenolic compounds . | FUNGAL BIOLOGY , 2023 , 127 (1-2) , 872-880 . |
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The medium-temperature alpha-amylase of Bacillus amyloliquefaciens is widely used in the food and washing process. Enhancing the thermostability of alpha-amylases and investigating the mechanism of stability are important for enzyme industry development. The optimal temperature and pH of the wild-type BAA and mutant MuBAA (D28E/V118A/S187D/K370 N) were all 60 degrees C and 6.0, respectively. The mutant MuBAA showed better thermostability at 50 degrees C and 60 degrees C, with a specific activity of 206.61 U/mg, which was 99.1% greater than that of the wild-type. By analyzing predicted structures, the improving thermostability of the mutant MuBAA was mainly related to enhanced stabilization of a loop region in domain B via more calcium-binding sites and intramolecular interactions around Asp187. Furthermore, additional intramolecular interactions around sites 28 and 370 in domain A were also beneficial for improving thermostability. Additionally, the decrease of steric hindrance at the active cavity increased the specific activity of the mutant MuBAA. Improving the thermostability of BAA has theoretical refer-ence values for the modification of alpha-amylases. (c) 2023 Elsevier Inc. All rights reserved.
Keyword ：
Alpha-amylase Alpha-amylase Bacillus amyloliquefaciens Bacillus amyloliquefaciens Thermostability Thermostability Three-dimensional model Three-dimensional model
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| GB/T 7714 | Yuan, Susu , Yan, Renxiang , Lin, Biyu et al. Improving thermostability of Bacillus amyloliquefaciens alpha-amylase by multipoint mutations [J]. | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS , 2023 , 653 : 69-75 . |
| MLA | Yuan, Susu et al. "Improving thermostability of Bacillus amyloliquefaciens alpha-amylase by multipoint mutations" . | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 653 (2023) : 69-75 . |
| APA | Yuan, Susu , Yan, Renxiang , Lin, Biyu , Li, Renkuan , Ye, Xiuyun . Improving thermostability of Bacillus amyloliquefaciens alpha-amylase by multipoint mutations . | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS , 2023 , 653 , 69-75 . |
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Bacillus amyloliquefaciens alpha-amylase (BAA) is one of well-known midrange thermostability amylases that are widely used in food and washing processes. However, the improvement of its catalytic properties by molecular modification is still lagging. To improve the activity of alpha-amylase BAA, mutants BAA28 and BAA294 are constructed via error-prone PCR and purified by column chromatography in the present study. The catalytic efficiencies (K-cat/K-m) of BAA28 and BAA294 are 2.42 and 2.73 mL mg(-1) s(-1), which are 43% and 61% higher than that of the wild-type BAA, respectively. Their specific activities are also increased by 40% and 62%, respectively, with no apparent changes of optimum temperature and pH. Homology modeling and molecular docking analysis suggest that the reduced steric hindrance is an important factor that enhances catalytic efficiencies and specific activities of the variants. These results may deepen the understanding of the mechanisms underlying the effects of each mutation on the catalytic efficiency of BAA and facilitate the construction of potent BAA mutants.
Keyword ：
Bacillus amyloliquefaciens alpha-amylase Bacillus amyloliquefaciens alpha-amylase protein structure protein structure random mutation random mutation specific activity specific activity
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| GB/T 7714 | Yuan, Susu , Li, Renkuan , Lin, Biyu et al. Engineering of Bacillus amyloliquefaciens α-Amylase for Improved Catalytic Efficiency by Error-Prone PCR [J]. | STARCH-STARKE , 2023 , 75 (11-12) . |
| MLA | Yuan, Susu et al. "Engineering of Bacillus amyloliquefaciens α-Amylase for Improved Catalytic Efficiency by Error-Prone PCR" . | STARCH-STARKE 75 . 11-12 (2023) . |
| APA | Yuan, Susu , Li, Renkuan , Lin, Biyu , Yan, Renxiang , Ye, Xiuyun . Engineering of Bacillus amyloliquefaciens α-Amylase for Improved Catalytic Efficiency by Error-Prone PCR . | STARCH-STARKE , 2023 , 75 (11-12) . |
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在pH8.5、37℃、酶与底物质量比0.5％的条件下用胰蛋白酶水解乳清分离蛋白(WPI)5 h制备水解物(WPH).通过控制环境条件,即添加不同盐(NaCl、KCl、CaCl2、MgCl2),改变离子强度(0.1 mol/L和1 mol/L),pH值(3～7)以及热处理来调控O/W界面处肽的组成.采用质谱法鉴定肽的组成,激光衍射法测量乳液的粒度分布及STEP技术测定乳液的稳定性,从而评估界面处肽的组成特性及相互作用对WPH乳化性的影响.结果表明,pH3时,因离子强度增大而引起的电荷屏蔽作用导致WPH乳液发生显著聚集,而pH7时离子强度对WPH乳化性的影响不显著,热处理则显著降低了其乳化性.此外,在WPH等电点(pH4)时,热处理显著提高了WPH的乳化性,尤其是添加CaCl2的样品组.综合不同pH条件下析出肽的组成及乳化性分析,研究发现来自β-Lg的肽段f(41～60)、f(71～91)、f(78～101)对WPH的乳化特性有重要贡献.
Keyword ：
pH值 pH值 乳化性 乳化性 乳清蛋白水解物 乳清蛋白水解物 热处理 热处理 盐离子 盐离子
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| GB/T 7714 | 黄雯婷 , 茅宇虹 , 蔡珍玲 et al. 不同条件下乳清蛋白水解物乳化性的变化及机理分析 [J]. | 中国食品学报 , 2023 , 23 (3) : 99-110 . |
| MLA | 黄雯婷 et al. "不同条件下乳清蛋白水解物乳化性的变化及机理分析" . | 中国食品学报 23 . 3 (2023) : 99-110 . |
| APA | 黄雯婷 , 茅宇虹 , 蔡珍玲 , 陈兰 , 李仁宽 , 叶秀云 . 不同条件下乳清蛋白水解物乳化性的变化及机理分析 . | 中国食品学报 , 2023 , 23 (3) , 99-110 . |
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Laccase, a multicopper oxidase, is well known for its industrial potentials to remove environmental pollutants due to its low substrate specificity to oxidize phenols and thus catalytic versatility. Many efforts focused on the metabolic mechanism, yet to decipher the structural determinants responsible for the differentiation between substrates. Aflatoxin B1 (AFB1), a new substrate for laccase, is a mycotoxin with a formidable environmental threat to public health and food safety. In the present study, we combined biochemical, in silico mutational and molecular-docking data to gain an insight to the function of key residues in the active cavity close to the T1 copper site in a characterized recombinant laccase from Cerrena unicolor (rCuL). Kinetic data for computerassisted virtual mutants established the binding affinity of hydrogen bonds and residues (Asn336, Asp207, Val391, and Thr165) in rCuL to AFB1. The augmented binding affinity to AFB1 may be related to the conformational rearrangements of the laccase and its ability to hydrogen-bond with the substrate. Furthermore, the optimal pH and temperature for rCuL and variants mediated AFB1 degradation may depend on their pH stability and thermostability. Our findings reinforce the importance of the structure-function relationship of fungal laccases in degrading AFB1, providing mechanistic guidance for future biocatalyst and bioengineering applications.
Keyword ：
Affinity Affinity Aflatoxin B 1 Aflatoxin B 1 Degradation Degradation Laccase Laccase Mutation Mutation
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| GB/T 7714 | Zhou, Zhimin , Li, Renkuan , Ng, Tzi Bun et al. Considerations regarding affinity determinants for aflatoxin B-1 in binding cavity of fungal laccase based on in silico mutational and verification studies [J]. | ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY , 2022 , 234 . |
| MLA | Zhou, Zhimin et al. "Considerations regarding affinity determinants for aflatoxin B-1 in binding cavity of fungal laccase based on in silico mutational and verification studies" . | ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 234 (2022) . |
| APA | Zhou, Zhimin , Li, Renkuan , Ng, Tzi Bun , Huang, Fang , Ye, Xiuyun . Considerations regarding affinity determinants for aflatoxin B-1 in binding cavity of fungal laccase based on in silico mutational and verification studies . | ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY , 2022 , 234 . |
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Microbial antibiotic resistance has become a worldwide concern, as it weakens the efficiency of the control of pathogenic microbes in both the fields of medicine and plant protection. A better understanding of antibiotic resistance mechanisms is helpful for the development of efficient approaches to settle this issue. In the present study, GC-MS-based metabolomic analysis was applied to explore the mechanisms of Zhongshengmycin (ZSM) resistance in Xanthomonas oryzae (Xoo), a bacterium that causes serious disease in rice. Our results show that the decline in the pyruvate cycle (the P cycle) was a feature for ZSM resistance in the metabolome of ZSM-resistant strain (Xoo-ZSM), which was further demonstrated as the expression level of genes involved in the P cycle and two enzyme activities were reduced. On the other hand, alanine was considered a crucial metabolite as it was significantly decreased in Xoo-ZSM. Exogenous alanine promoted the P cycle and enhanced the ZSM-mediated killing efficiency in Xoo-ZSM. Our study highlights that the depressed P cycle is a feature in Xoo-ZSM for the first time. Additionally, exogenous alanine is a candidate enhancer and can be applied with ZSM to improve the antibiotic-mediated killing efficiency in the control of infection caused by Xoo.
Keyword ：
alanine alanine antibiotic resistance antibiotic resistance P cycle P cycle Xanthomonas oryzae Xanthomonas oryzae Zhongshengmycin Zhongshengmycin
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| GB/T 7714 | Guan, Yi , Shen, Peihua , Lin, Meiyun et al. Exogenous Alanine Reverses the Bacterial Resistance to Zhongshengmycin with the Promotion of the P Cycle in Xanthomonas oryzae [J]. | ANTIBIOTICS-BASEL , 2022 , 11 (2) . |
| MLA | Guan, Yi et al. "Exogenous Alanine Reverses the Bacterial Resistance to Zhongshengmycin with the Promotion of the P Cycle in Xanthomonas oryzae" . | ANTIBIOTICS-BASEL 11 . 2 (2022) . |
| APA | Guan, Yi , Shen, Peihua , Lin, Meiyun , Ye, Xiuyun . Exogenous Alanine Reverses the Bacterial Resistance to Zhongshengmycin with the Promotion of the P Cycle in Xanthomonas oryzae . | ANTIBIOTICS-BASEL , 2022 , 11 (2) . |
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为阐明BbRho5对球孢白僵菌生防潜能的作用,构建了Bbrho5单基因敲除菌株ΔBbrho5,以野生型菌株WT作为对照,在不同培养基上测定菌落生长速率,并测定了菌株对多菌灵胁迫耐受性及对大蜡螟幼虫体壁侵染能力.进一步获取和分析了ΔBbrho5和WT细胞内基因转录组数据.结果表明,BbRho5蛋白功能缺陷显著抑制球孢白僵菌菌丝生长速率,同时微弱影响其多菌灵胁迫抗逆性及生防能力.相较于WT,ΔBbrho5中具有770个差异表达基因(DEGs),其中上调基因395个,下调基因375个.GO分析显示,ΔBbrho5 VS WT中DEGs主要富集于氧化还原酶活力(oxidoreductase activity)和单加氧酶活力(monooxygenase activity)功能.KEGG通路富集结果显示,DEGs主要富集于氮代谢及多种氨基酸代谢通路.在氮代谢通路中富集到7个功能基因,其中有5个上调,2个下调,说明敲除菌株可能采用增强氮源利用及谷氨酸合成以应对Bbrho5缺陷引起的生长迟缓.以上研究结果揭示了球孢白僵菌中小GTP酶BbRho5对球孢白僵菌生长速率具有重要影响,且氮代谢和氨基酸代谢可能为其重要的响应代谢通路.
Keyword ：
BbRho5 BbRho5 小GTP酶 小GTP酶 球孢白僵菌 球孢白僵菌 生长速率 生长速率 转录组 转录组
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| GB/T 7714 | 关怡 , 李新 , 王定一 et al. BbRho5对球孢白僵菌生长速率的作用研究 [J]. | 生物技术通报 , 2022 , 38 (2) : 132-140 . |
| MLA | 关怡 et al. "BbRho5对球孢白僵菌生长速率的作用研究" . | 生物技术通报 38 . 2 (2022) : 132-140 . |
| APA | 关怡 , 李新 , 王定一 , 杜茜 , 张龙斌 , 叶秀云 . BbRho5对球孢白僵菌生长速率的作用研究 . | 生物技术通报 , 2022 , 38 (2) , 132-140 . |
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This study aimed to improve the stability and catalytic properties of Thermomyces lanuginosus lipase (TLL) adsorbed on a hydrophobic support. At the optimized conditions (pH 5 and 25 degrees C without any additions), the Sips isotherm model effectively fitted the equilibrium adsorption data, indicating a monolayer and the homogenous distribution of immobilized lipase molecules. To preserve the high specific activity of adsorbed lipase, the immobilized lipase (IL) with a moderate loading amount (approximately 40% surface coverage) was selected. Polyethylenimine (PEI) and chitosan (CS) were successfully applied as bridging units to in situ crosslink the immobilized lipase molecules in IL. At the low polymer concentration (0.5%, w/w) and with 1 h incubation, insignificant changes in average pore size were detected. Short-chain PEI and CS (MW <= 2 kDa) efficiently improved the lipase stability, i.e., the lipase loss decreased from 40% to <2%. Notably, CS performed much better than PEI in maintaining lipase activity. IL crosslinked with CS-2 kDa showed a two- to three-fold higher rate when hydrolyzing p-nitrophenyl butyrate and a two-fold increase in the catalytic efficiency in the esterification of hexanoic acid with butanol. These in situ crosslinking strategies offer good potential for modulating the catalytic properties of TLL for a specific reaction.
Keyword ：
anti-desorption anti-desorption chitosan chitosan esterification esterification hydrolytic activity hydrolytic activity polyethylenimine polyethylenimine
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| GB/T 7714 | Mao, Yuhong , Cai, Zhenling , Zhou, Chenxi et al. Characteristics of Crosslinking Polymers Play Major Roles in Improving the Stability and Catalytic Properties of Immobilized Thermomyces lanuginosus Lipase [J]. | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2022 , 23 (6) . |
| MLA | Mao, Yuhong et al. "Characteristics of Crosslinking Polymers Play Major Roles in Improving the Stability and Catalytic Properties of Immobilized Thermomyces lanuginosus Lipase" . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 23 . 6 (2022) . |
| APA | Mao, Yuhong , Cai, Zhenling , Zhou, Chenxi , Lan, Hangzhen , Ye, Xiuyun . Characteristics of Crosslinking Polymers Play Major Roles in Improving the Stability and Catalytic Properties of Immobilized Thermomyces lanuginosus Lipase . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2022 , 23 (6) . |
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