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Screening of lipase TiL from Tilletia indica for chemo-enzymatic epoxidation of alkenes 筛选蕉叶中用于烯烃化学酶促环氧化的脂肪酶 TiL
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-22 DOI: 10.1016/j.enzmictec.2024.110547
Jiang Pan , Nan Yang , Yuan-Lin Lv , Zi-Yang Zhang , Chun-Xiu Li, Jian-He Xu
Lipase can mediate the chemo-enzymatic epoxidation of alkenes with the presence of free carboxylic acid and hydrogen peroxide. Four novel lipases with the abilities of chemo-enzymatic epoxidation were mined from the gene database. Lipase TiL originated from Tilletia indica was identified with significant activity on formation of methyl epoxystearate from methyl oleate. n-Heptanoic acid was determined as the optimal carboxylic acid substrate of TiL. Methyl oleate and α-pinene were efficiently converted to corresponding epoxy compound in micro-aqueous media and aqueous-organic biphase, respectively. A preparative scale chemo-enzymatic transformation of α-pinene was conduct using the optimized reaction condition, with 30 % yield of α-pinene oxide obtained.
脂肪酶可以在存在游离羧酸和过氧化氢的情况下介导烯烃的化学酶促环氧化反应。我们从基因数据库中挖掘出四种具有化学酶促环氧化能力的新型脂肪酶。确定了正庚酸为 TiL 的最佳羧酸底物。在微水介质和水有机双相中,油酸甲酯和 α-蒎烯分别被高效地转化为相应的环氧化合物。利用优化的反应条件对α-蒎烯进行了制备规模的化学酶转化,获得了 30% 的α-蒎烯氧化物收率。
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引用次数: 0
Improvement of lipid production from glucose/xylose mixed-sugar by the oleaginous yeast Lipomyces starkeyi through ultra-violet mutagenesis 通过紫外线诱变提高含油酵母星形脂酵母从葡萄糖/木糖混合糖中产生脂质的能力
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-22 DOI: 10.1016/j.enzmictec.2024.110551
Sota Kamba, Ryosuke Yamada , Takuya Matsumoto, Hiroyasu Ogino
The oleaginous yeast Lipomyces starkeyi is a promising triacylglycerol (TAG) producer for biodiesel fuel. However, it is necessary to further improve TAG productivity in L. starkeyi from a mixed sugar of glucose and xylose. This study aimed to construct an L. starkeyi mutant with increased TAG productivity from glucose/xylose mixed-sugar and to elucidate the causes underlying increased lipid productivity. Ultra-violet (UV) mutagenesis combined with enrichment culture with ethanol and H2O2 and selection of low-density cells was applied to L. starkeyi to obtain the L. starkeyi mutant strain UMP47, which exhibited higher TAG production from glucose/xylose. Transcriptome analysis revealed high expression of genes involved in transporter activity and carbohydrate metabolism, whereas genes involved in DNA replication exhibited lower expression in the mutant strain UMP47 than in the wild-type strain. Altogether, the lipid productivity of L. starkeyi was successfully improved by UV mutagenesis. Transcriptome analysis suggested the importance of previously unidentified genes in TAG production. This study provides information on potential target genes for improving TAG production through the genetic modification of oleaginous yeast.
含油酵母星形脂酵母是一种很有前途的生物柴油燃料三酰甘油(TAG)生产者。然而,有必要进一步提高星状酵母从葡萄糖和木糖混合糖中生产三酰甘油(TAG)的能力。本研究旨在构建一种可提高葡萄糖/木糖混合糖 TAG 生产率的星菌突变体,并阐明提高脂质生产率的原因。通过紫外线(UV)诱变、乙醇和 H2O2 富集培养以及低密度细胞的筛选,我们获得了星形菌突变株 UMP47,该突变株从葡萄糖/木糖中获得了更高的 TAG 产量。转录组分析显示,涉及转运体活性和碳水化合物代谢的基因表达量较高,而涉及 DNA 复制的基因在突变株 UMP47 中的表达量低于野生型菌株。总之,紫外诱变成功地提高了L. starkeyi的脂质生产率。转录组分析表明,以前未发现的基因在 TAG 生产中起着重要作用。这项研究为通过对含油酵母进行基因改造来提高 TAG 产量提供了潜在目标基因的信息。
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引用次数: 0
Affordable infectious pathogen detection using a dual-mode biosensor integrating exonuclease III-assisted target recycling amplification with high-throughput 96-well microplate format 使用双模式生物传感器检测传染病病原体,该传感器集成了外切酶 III 辅助目标循环扩增和高通量 96 孔微孔板格式,价格低廉
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-21 DOI: 10.1016/j.enzmictec.2024.110549
Hamza Moustakim, Aziz Amine, Hasna Mohammadi
The ongoing challenge of infectious pathogens highlights the need for accurate and accessible methods to discern their genetic signatures, especially in resource-limited settings. In response to this crucial requirement, we introduce an affordable large-scale screening platform for infectious pathogen detection, using Hepatitis B virus (HBV) as a fundamental model. This proposed biosensor integrates an exonuclease III-assisted target recycling amplification strategy within a high-throughput 96-well microplate format. The HBV DNA target binds to a capture probe DNA and exonuclease III digests the probe to release the target. This mechanism enables the target to engage in binding cycles with new probes, each digested in turn, increasing detection sensitivity for even small quantities of HBV DNA. The implemented approach incorporates a biotin-streptavidin interaction allowing the undigested capture probe DNA to bind to a 5′-biotin-modified detection probe for effective HBV DNA quantification. This interaction generates a signal that, following the enzyme-substrate reaction, can be detected on-site using a smartphone, offering either optical or electrochemical readouts. The developed biosensor was capable of detecting HBV DNA with a detection limit of 5.62 fM and provided a considerable linear range covering concentrations from 100 fM to 100 nM. The determination of HBV DNA quantities in spiked human serum was achieved with a recovery of 90.0 % – 107.4 % as well. The results suggest that the developed dual-mode biosensor offers an adaptable and cost-effective approach for detecting infectious diseases, with promising applications in medical diagnostics and environmental monitoring to support public health efforts.
传染病病原体带来的持续挑战凸显了对准确、易用的方法的需求,尤其是在资源有限的环境中。针对这一关键需求,我们以乙型肝炎病毒(HBV)为基本模型,推出了一种经济实惠的大规模传染病病原体检测筛查平台。这种拟议的生物传感器在高通量 96 孔微孔板格式中整合了外切酶 III 辅助的靶循环扩增策略。HBV DNA 靶标与捕获探针 DNA 结合,外切酶 III 消化探针以释放靶标。这种机制能使目标与新探针循环结合,每个探针依次被消化,从而提高对少量 HBV DNA 的检测灵敏度。这种方法结合了生物素-链霉亲和素的相互作用,使未消化的捕获探针 DNA 与 5′-生物素修饰的检测探针结合,从而有效地定量检测 HBV DNA。在酶-底物反应后,这种相互作用产生的信号可通过智能手机现场检测,提供光学或电化学读数。所开发的生物传感器能够检测出检测限为 5.62 fM 的 HBV DNA,其线性范围相当大,可覆盖 100 fM 到 100 nM 的浓度范围。在测定加标人血清中的 HBV DNA 数量时,回收率也达到了 90.0 % - 107.4 %。结果表明,所开发的双模式生物传感器为检测传染病提供了一种适应性强、成本效益高的方法,有望应用于医疗诊断和环境监测,为公共卫生工作提供支持。
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引用次数: 0
An environmental “fairytail”: Removal of mercury from water via phage virion-based biosorption 环境 "仙女棒":通过基于噬菌体病毒的生物吸附技术去除水中的汞。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-19 DOI: 10.1016/j.enzmictec.2024.110548
Larissa F. Santos , Denicezar Â. Baldo , José M. Oliveira Jr , Marta M.D.C. Vila , Victor M. Balcão
Contamination of water with mercury constitutes a serious public health problem, especially in locations where the use of Hg occurs improperly/illegally and negligently, as is the case in the Amazon region (Brazil). The riverside populations in the Amazon are frequently invaded by illegal mining, exposing these populations to significant risks, of which contamination by heavy metals such as mercury (Hg2+) has the potential to cause serious illnesses. Furthermore, exposure to this metal causes neurological, cardiovascular, immune and digestive system disorders, in addition to damaging the lungs, kidneys, skin and eyes. The aquatic biome is extremely important for the local economy and population, being drastically affected by Hg2+ contamination and its effects. Therefore, it is necessary to develop bioremediation/biomitigation methods that are effective and less harmful to the environment, aiming to remove Hg2+ from water. Hence, when we think about new methodologies that can lead to the reduction of mercury in water, the use of protein entities is a potential option and, for this reason, we can highlight the possibility of using bacteriophage virions to remove Hg2+ ions from water by biosorption using their negative Zeta Potential for this purpose. In this sense, the main goal of the research work undertaken was to test the possibility of mitigating the presence of mercury (II) ions in water through the immobilization of a bacteriophage isolated and already characterized by our research group (EcoM021, T4 myovirus of the Straboviridae family and genus Tequatrovirus), on a chitosan-coated Ca-alginate microparticle support, through which water contaminated with Hg2+ ions was percolated. The system developed in microparticle form integrating trapped phage virions showed to be very promising for retaining mercury ions through biosorption (electrostatic attraction), thus enabling the removal of ionic mercury from water.
汞对水的污染是一个严重的公共卫生问题,尤其是在汞的使用不当/非法和疏忽的地 方,亚马逊地区(巴西)就是这种情况。亚马逊河流域的居民经常受到非法采矿的侵扰,使这些居民面临巨大的风险,其中汞(Hg2+)等重金属的污染有可能导致严重的疾病。此外,接触这种金属会导致神经、心血管、免疫和消化系统紊乱,还会损害肺、肾、皮肤和眼睛。水生生物群落对当地经济和人口极为重要,受到 Hg2+ 污染及其影响的严重影响。因此,有必要开发有效且对环境危害较小的生物修复/生物缓解方法,以去除水中的 Hg2+。因此,当我们考虑可以减少水中汞含量的新方法时,使用蛋白质实体是一个潜在的选择,为此,我们可以强调利用噬菌体病毒的负 Zeta 电位,通过生物吸附去除水中 Hg2+ 离子的可能性。从这个意义上说,这项研究工作的主要目标是通过在壳聚糖包覆的 Ca-alginate 微粒支撑物上固定我们研究小组分离并已定性的噬菌体(EcoM021,Straboviridae 科、Tequatrovirus 属的 T4 myovirus),测试减轻水中汞 (II) 离子存在的可能性。通过生物吸附(静电吸引)保留汞离子,从而去除水中的离子汞。
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引用次数: 0
Enhanced lipid accumulation in microalgae Scenedesmus sp. under nitrogen limitation 氮限制条件下微藻类 Scenedesmus sp.
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-09 DOI: 10.1016/j.enzmictec.2024.110546
Getachew Tafere Abrha , Abdalah Makaranga , Pannaga Pavan Jutur
Microalgae-based biofuel production is cost-effective only in a biorefinery, where valuable co-products offset high costs. Fatty acids produced by photosynthetic microalgae can serve as raw materials for bioenergy and pharmaceuticals. This study aims to understand the metabolic imprints of Scenedesmus sp. CABeR52, to decipher the physiological mechanisms behind lipid accumulation under nitrogen deprivation. Metabolomics profiles were generated using gas chromatography-mass spectrometry (GC–MS) of Scenedesmus sp. CABeR52 subjected to nutrient deprivation. Our initial data sets indicate that deprived cells have an increased accumulation of lipids (278.31 mg.g−1 dcw), 2.0 times higher than the control. The metabolomic profiling unveils a metabolic reprogramming, highlighting the upregulation of key metabolites involved in fatty acid biosynthesis, such as citric acid, succinic acid, and 2-ketoglutaric acid. The accumulation of trehalose, a stress-responsive metabolite, further underscores the microalga's adaptability. Interestingly, we found that a new fatty acid, nervonic acid, was identified in the complex, which has a significant role in brain development. These findings provide valuable insights into the metabolic pathways governing lipid accumulation in Scenedesmus sp., paving the way for its exploitation as a sustainable biofuel feedstock.
只有在生物精炼厂中,基于微藻的生物燃料生产才具有成本效益,因为在生物精炼厂中,有价值的副产品可以抵消高昂的成本。光合微藻产生的脂肪酸可以作为生物能源和制药的原料。本研究旨在了解 Scenedesmus sp. CABeR52 的代谢印记,以破译缺氮条件下脂质积累背后的生理机制。研究人员利用气相色谱-质谱联用技术(GC-MS)生成了营养匮乏条件下的景天科植物 CABeR52 的代谢组学图谱。我们的初步数据集表明,缺乏营养的细胞中脂类积累增加(278.31 mg.g-1 dcw),是对照组的 2.0 倍。代谢组分析揭示了一种代谢重编程,突出显示了参与脂肪酸生物合成的关键代谢物的上调,如柠檬酸、琥珀酸和 2-酮戊二酸。应激反应代谢物三卤糖的积累进一步突出了微藻的适应性。有趣的是,我们发现在该复合体中发现了一种新的脂肪酸--神经酸,它在大脑发育中起着重要作用。这些发现为我们深入了解管理景天藻脂质积累的代谢途径提供了宝贵的信息,为将其开发为可持续生物燃料原料铺平了道路。
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引用次数: 0
Biochemical characterization and structure prediction of the Cerrado soil CRB2(1) metagenomic dioxygenase Cerrado土壤CRB2(1)元基因组二氧酶的生化特征和结构预测。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-08 DOI: 10.1016/j.enzmictec.2024.110544
Philippe de Castro Lins , Pedro Ricardo Vieira Hamann , Jônatas Cunha Barbosa Lima , João Alexandre Ribeiro Gonçalves Barbosa , João Lucas da Silva Correia , Ikaro Alves de Andrade , Débora Farage Knupp dos Santos , Betania Ferraz Quirino , Ricardo Henrique Krüger
Dioxygenases are enzymes involved in the conversion of polyconic aromatic hydroxycarbons (PAHs), attracting significant biotechnological interest for the conversion of recalcitrant organic compounds. Furthermore, few studies show that dioxygenases can take on the function of resistance genes in clones. This enzymatic versatility opens up new opportunities for elucidating the mechanisms of microbial resistance, as well as its biotechnological application. In this work, a Cerrado soil dioxygenase named CRB2(1) was biochemically characterized. The enzyme was shown to have optimal activity at pH 7; a temperature of 30 °C; and using iron ions as a cofactor for substrate cleavage. The kinetic catalytic parameters of CRB2(1) were Vmax = 0.02281 µM/min and KM = 97.6. Its predicted three-dimensional structure obtained using the Modeller software v9.22 based on the crystal structure of gentisate 1,2-dioxygenase from Silicibacter pomeroyi (GDOsp) (PDB ID 3BU7, resolution 2.80 Å, residues 17–374) revealed substrate binding to the cupin domain, where the active site is located. The analyzed substrates interact directly with the iron ion, coordinated by three histidine residues. Changing the iron ion charge modifies the binding between the active site and the substrates. Currently, there is a demand for enzymes that have biotechnological activities of interest. Metagenomics allows analyzing the biotechnological potential of several organisms at the same time, based on sequence and functional activity analyses.
二氧酶是一种参与多环芳烃(PAHs)转化的酶,在转化难降解有机化合物方面引起了生物技术的极大兴趣。此外,很少有研究表明二氧酶可以在克隆中承担抗性基因的功能。这种酶的多功能性为阐明微生物抗性机理及其生物技术应用提供了新的机遇。在这项工作中,对一种名为 CRB2(1) 的 Cerrado 土壤二氧酶进行了生物化学鉴定。研究表明,该酶在 pH 值为 7、温度为 30 °C、以铁离子作为辅助因子裂解底物时具有最佳活性。CRB2(1) 的动力学催化参数为 Vmax = 0.02281 µM/min 和 KM = 97.6。根据波美拉尼亚硅杆菌(Silicibacter pomeroyi)的龙胆二酸 1,2-二氧合酶(GDOsp)晶体结构(PDB ID 3BU7,分辨率 2.80 Å,残基 17-374),使用 Modeller 软件 v9.22 对其三维结构进行了预测,结果显示底物与活性位点所在的杯状结构域结合。分析的底物直接与由三个组氨酸残基配位的铁离子相互作用。改变铁离子的电荷会改变活性位点与底物之间的结合。目前,人们需要具有生物技术活性的酶。元基因组学可以根据序列和功能活性分析,同时分析几种生物的生物技术潜力。
{"title":"Biochemical characterization and structure prediction of the Cerrado soil CRB2(1) metagenomic dioxygenase","authors":"Philippe de Castro Lins ,&nbsp;Pedro Ricardo Vieira Hamann ,&nbsp;Jônatas Cunha Barbosa Lima ,&nbsp;João Alexandre Ribeiro Gonçalves Barbosa ,&nbsp;João Lucas da Silva Correia ,&nbsp;Ikaro Alves de Andrade ,&nbsp;Débora Farage Knupp dos Santos ,&nbsp;Betania Ferraz Quirino ,&nbsp;Ricardo Henrique Krüger","doi":"10.1016/j.enzmictec.2024.110544","DOIUrl":"10.1016/j.enzmictec.2024.110544","url":null,"abstract":"<div><div>Dioxygenases are enzymes involved in the conversion of polyconic aromatic hydroxycarbons (PAHs), attracting significant biotechnological interest for the conversion of recalcitrant organic compounds. Furthermore, few studies show that dioxygenases can take on the function of resistance genes in clones. This enzymatic versatility opens up new opportunities for elucidating the mechanisms of microbial resistance, as well as its biotechnological application. In this work, a <em>Cerrado</em> soil dioxygenase named CRB2(1) was biochemically characterized. The enzyme was shown to have optimal activity at pH 7; a temperature of 30 °C; and using iron ions as a cofactor for substrate cleavage. The kinetic catalytic parameters of CRB2(1) were <em>V</em><sub>max</sub> = 0.02281 µM/min and <em>K</em><sub>M</sub> = 97.6. Its predicted three-dimensional structure obtained using the Modeller software v9.22 based on the crystal structure of gentisate 1,2-dioxygenase from <em>Silicibacter pomeroyi</em> (GDOsp) (PDB ID <span><span>3BU7</span><svg><path></path></svg></span>, resolution 2.80 Å, residues 17–374) revealed substrate binding to the cupin domain, where the active site is located. The analyzed substrates interact directly with the iron ion, coordinated by three histidine residues. Changing the iron ion charge modifies the binding between the active site and the substrates. Currently, there is a demand for enzymes that have biotechnological activities of interest. Metagenomics allows analyzing the biotechnological potential of several organisms at the same time, based on sequence and functional activity analyses.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110544"},"PeriodicalIF":3.4,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142617164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Probiotic and functional characterization of newly isolated Lactiplantibacillus plantarum strains from human breast milk and proliferative inhibition potential of metabolites 从母乳中新分离出的植物乳杆菌菌株的益生特性和功能以及代谢产物的增殖抑制潜力。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-07 DOI: 10.1016/j.enzmictec.2024.110545
Yusuf Alan , Ali-Osman Keskin , Mehmet Sönmez
Four Lactiplantibacillus plantarum strains newly isolated and identified from human breast milk in Türkiye, have probiotic, functional and proliferative inhibition potential of metabolites against colon cancer cell lines were evaluated. In simulated gastric and intestinal media, all strains exhibited strong probiotic character by showing resistance, although decreasing with time and concentration. The strains were sensitive to penicillin G, rifampin and chloramphenicol and showed antibacterial effect on all pathogenic bacteria. Citric acid, malic acid, tartaric acid, pyruvic acid and fumaric acid were not detected in the strains, while the highest amount of acetic acid was detected. The quantitative-qualitative analysis and structural characterization of exopolysaccharide (EPS) was confirmed and it was determined that the strains synthesized similar amounts. Compared to standard antioxidants, the strains showed less DPPH activity and similar ABTS activity. High amounts of metabolites of the strains showed good antiproliferative effect on Caco-2, while lower amounts showed good antiproliferative effect on the HT-29 cell line. When all the data were considered, it was determined that the strains were close to each other, but the YAAS 23 strain showed slightly better properties. In conclusion, breast milk is a unique environment harboring beneficial bacteria such as L. plantarum for human health.
研究人员评估了从土耳其人的母乳中新分离和鉴定出的四株植物乳杆菌(Lactiplantibacillus plantarum)对结肠癌细胞株的益生、功能和代谢物增殖抑制潜力。在模拟胃和肠道培养基中,所有菌株都表现出很强的益生特性,表现出抗药性,但随着时间和浓度的增加而降低。这些菌株对青霉素 G、利福平和氯霉素敏感,对所有致病菌都有抗菌作用。菌株中未检测到柠檬酸、苹果酸、酒石酸、丙酮酸和富马酸,而乙酸的含量最高。外多糖(EPS)的定量-定性分析和结构特征得到证实,并确定菌株合成的外多糖数量相似。与标准抗氧化剂相比,菌株的 DPPH 活性较低,ABTS 活性相似。高含量的菌株代谢物对 Caco-2 细胞系具有良好的抗增殖作用,而低含量的菌株代谢物对 HT-29 细胞系具有良好的抗增殖作用。综合所有数据,可以确定这些菌株的性能接近,但 YAAS 23 菌株的性能略胜一筹。总之,母乳是一个独特的环境,其中蕴藏着植物乳杆菌等对人体健康有益的细菌。
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引用次数: 0
Poly-γ-glutamic production by solid-state fermentation of Bacillus natto in ammonia nitrogen movement and soil water retention processes 纳豆芽孢杆菌固态发酵在氨氮移动和土壤保水过程中产生聚-γ-谷氨酸。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-31 DOI: 10.1016/j.enzmictec.2024.110543
Xin Wang , Jie Gao , Jiahui Wu , Xuan Li , Junxun Li , Haihong Li , Songlin Wang
A high polyglutamic acid (γ-PGA) producing strain of Bacillus natto UV-40–50 was screened by ultraviolet mutagenesis treatment and identified as still belonging to the Bacillus specie. The optimal fermentation medium composition for solid state fermentation (SSF) of B. natto strain UV-40–50 strain was determined by one-way analysis of variance, under which the yield of γ-PGA was 55.19 g/kg, and the presence and molecular weight of γ-PGA in the γ-PGA-purified samples were determined by a series of characterizations. The purification ability of the unseparated solid fermentation product (SFP) on ammonia nitrogen and nitrite in the water column, as well as its effect on soil water retention, germination rate and seedling length of lettuce and cabbage were further investigated. The results showed that the addition of 1 g/m3 SFP could effectively remove more than 60 % of ammonia nitrogen and more than 40 % of nitrite in the water body; the addition of 0.01 % SFP could increase the water retention capacity of cabbage soil by 2.13 times, and increase the water retention capacity of lettuce soil by 12 %; at the same time, the SFP could also significantly increase the germination rate and seedling length of both cabbage and lettuce.
通过紫外线诱变处理筛选出一株高产聚谷氨酸(γ-PGA)的纳豆芽孢杆菌 UV-40-50 株,并确定其仍属于芽孢杆菌属。通过单因素方差分析确定了纳豆芽孢杆菌 UV-40-50 菌株固态发酵(SSF)的最佳发酵培养基组成,在此条件下,γ-PGA 的产量为 55.19 g/kg,并通过一系列表征确定了γ-PGA 纯化样品中γ-PGA 的存在和分子量。进一步研究了未分离固体发酵产物(SFP)对水体中氨氮和亚硝酸盐的净化能力,以及对土壤保水性、莴苣和卷心菜发芽率和苗长的影响。结果表明,添加 1 克/立方米的 SFP 能有效去除水体中 60% 以上的氨氮和 40% 以上的亚硝酸盐;添加 0.01% 的 SFP 能使白菜土壤的保水能力提高 2.13 倍,使莴苣土壤的保水能力提高 12%;同时,SFP 还能显著提高白菜和莴苣的发芽率和苗长。
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引用次数: 0
Advances and prospects for lactic acid production from lignocellulose 利用木质纤维素生产乳酸的进展与前景。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1016/j.enzmictec.2024.110542
Ruofan Wu, Jiahui Yang, Yujia Jiang, Fengxue Xin
Lactic acid is a versatile building block that can be produced via microbial fermentation. Owing to the high optical purity, approximately 90 % of lactic acid is produced by microbes. Recently, the biosynthesis of lactic acid from lignocellulose has concerned much attentions. However, the cost-effective process faces several obstacles because of the complex structure of lignocellulose. This review will comprehensively summarize the state-of-the-art lactic acid production from lignocellulose, including the commonly used lactate-producing microorganisms, the co-utilization of glucose and xylose for the lactic acid production, as well as the lactic acid production from lignocellulose hydrolysate. Furthermore, the strategies regarding the lignocellulosic lactic acid production via consolidated bioprocessing will be also discussed, which can greatly reduce the complexity of the fermentation process.
乳酸是一种可通过微生物发酵生产的多功能建筑材料。由于光学纯度高,大约 90% 的乳酸由微生物生产。最近,从木质纤维素中生物合成乳酸备受关注。然而,由于木质纤维素结构复杂,这一经济有效的工艺面临着一些障碍。本综述将全面总结从木质纤维素中生产乳酸的最新技术,包括常用的乳酸生产微生物、葡萄糖和木糖共同用于乳酸生产以及从木质纤维素水解物中生产乳酸。此外,还将讨论通过综合生物工艺生产木质纤维素乳酸的策略,这可以大大降低发酵过程的复杂性。
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引用次数: 0
The synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid by a GDSL-type esterase and its substrate specificity analysis 一种 GDSL 型酯酶合成乙酸肉桂酯和脱乙酰基-7-氨基头孢烷酸及其底物特异性分析。
IF 3.4 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-28 DOI: 10.1016/j.enzmictec.2024.110532
Shuqi Xing , Wei Xie , Guangli Hu , Chaocheng Luo , Hong Zhu , Laping He , Cuiqin Li , Xiao Wang , Xuefeng Zeng
GDSL-type esterases are promising biocatalysts for the food and pharmaceutical industries. Here, a GDSL-type esterase from Aspergillus niger CCTCC No. M2012538 (INANE1) was expressed and purified in Pichia pastoris GS115, and its catalytic performances were evaluated, including the synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid (D-7-ACA). In addition, molecular docking and molecular dynamics simulations analyzed INANE1's substrate specificity. The substrate specificity profile indicated the recombinant esterase (rINANE1) was an acetylesterase with high specificity for p-nitrophenyl acetate (p-NPA). The rINANE1 exhibited maximum activity at pH 8.0 and 35 °C, where Km and Vmax were calculated as 0.13±0.03 mM and 22.56 ± 0.32 μmoL/min/mg, respectively. The yield of cinnamyl acetate of about 85 % was achieved in 24 h. The conversion rate of 7-aminocephalosporanic acid (7-ACA) could reach 92.71 ± 1.78 % at 25 °C and 2.5 h. Moreover, the INANE1 structure model, molecular docking, and molecular dynamics simulation demonstrated that the pocket of the catalytic triad Ser34, Asn267, and His270 could only accommodate p-NPA. INANE1 may be the first fungi esterase with cinnamyl acetate synthetic activity and 7-ACA hydrolysis activity. Therefore, INANE1 would be a promising enzyme with industrial values.
GDSL 型酯酶是一种在食品和制药行业很有前景的生物催化剂。本文在 Pichia pastoris GS115 中表达并纯化了黑曲霉 CCTCC 编号 M2012538 的 GDSL 型酯酶(INANE1),并对其催化性能进行了评估,包括乙酸肉桂酯和脱乙酰基-7-氨基头孢烷酸(D-7-ACA)的合成。此外,分子对接和分子动力学模拟分析了 INANE1 的底物特异性。底物特异性曲线表明,重组酯酶(rINANE1)是一种乙酰酯酶,对对硝基苯乙酸酯(p-NPA)具有高度特异性。rINANE1 在 pH 值为 8.0、温度为 35 ℃ 时表现出最大活性,其 Km 和 Vmax 分别为 0.13±0.03 mM 和 22.56 ± 0.32 μmoL/min/mg 。在 24 小时内,乙酸肉桂酯的收率达到约 85%。在 25 °C、2.5 h 的条件下,7-氨基头孢烷酸(7-ACA)的转化率可达 92.71 ± 1.78 %。此外,INANE1结构模型、分子对接和分子动力学模拟表明,催化三元组Ser34、Asn267和His270的口袋只能容纳对-NPA。INANE1 可能是第一个具有乙酸肉桂酯合成活性和 7-ACA 水解活性的真菌酯酶。因此,INANE1 将是一种具有工业价值的酶。
{"title":"The synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid by a GDSL-type esterase and its substrate specificity analysis","authors":"Shuqi Xing ,&nbsp;Wei Xie ,&nbsp;Guangli Hu ,&nbsp;Chaocheng Luo ,&nbsp;Hong Zhu ,&nbsp;Laping He ,&nbsp;Cuiqin Li ,&nbsp;Xiao Wang ,&nbsp;Xuefeng Zeng","doi":"10.1016/j.enzmictec.2024.110532","DOIUrl":"10.1016/j.enzmictec.2024.110532","url":null,"abstract":"<div><div>GDSL-type esterases are promising biocatalysts for the food and pharmaceutical industries. Here, a GDSL-type esterase from <em>Aspergillus niger</em> CCTCC No. M2012538 (INANE1) was expressed and purified in <em>Pichia pastoris</em> GS115, and its catalytic performances were evaluated, including the synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid (D-7-ACA). In addition, molecular docking and molecular dynamics simulations analyzed INANE1's substrate specificity. The substrate specificity profile indicated the recombinant esterase (rINANE1) was an acetylesterase with high specificity for <em>p</em>-nitrophenyl acetate (<em>p</em>-NPA). The rINANE1 exhibited maximum activity at pH 8.0 and 35 °C, where K<sub>m</sub> and V<sub>max</sub> were calculated as 0.13±0.03 mM and 22.56 ± 0.32 μmoL/min/mg, respectively. The yield of cinnamyl acetate of about 85 % was achieved in 24 h. The conversion rate of 7-aminocephalosporanic acid (7-ACA) could reach 92.71 ± 1.78 % at 25 °C and 2.5 h. Moreover, the INANE1 structure model, molecular docking, and molecular dynamics simulation demonstrated that the pocket of the catalytic triad Ser34, Asn267, and His270 could only accommodate <em>p</em>-NPA. INANE1 may be the first fungi esterase with cinnamyl acetate synthetic activity and 7-ACA hydrolysis activity. Therefore, INANE1 would be a promising enzyme with industrial values.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110532"},"PeriodicalIF":3.4,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Enzyme and Microbial Technology
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