耐高盐枯草芽孢杆菌XP合成球形纳米硒及其抑制草莓病原真菌生物活性

生物方法合成纳米材料具有低能耗、高安全性以及环境友好等优良特点,因而备受人们关注.利用细菌将硒酸盐或亚硒酸盐还原为单质硒,不仅可以降低硒毒性,而且还能获得价值更高的生物纳米材料.文中选用可耐受高盐环境胁迫的枯草芽孢杆菌亚种Bacillus subtilis subspecies stercoris strain XP构...     

利用副干酪乳酪杆菌SCFF20高效生物合成纳米硒颗粒：一种潜在的亚硒酸盐生物转化工厂

【目的】硒(Se)是人体必需的微量元素,在维持人体生理代谢中起着至关重要的作用。在硒的各种形态中,纳米硒颗粒(selenium nanoparticles, SeNPs)被发现具有较高的生物利用度和较低的毒性。本研究拟筛选一株能将亚硒酸盐高效合成纳米硒颗粒的益生菌菌株。【方法】从14株潜在益生菌中筛选出一株能有效将亚硒酸钠转化为SeNPs的耐硒菌株副干酪乳酪杆菌SCFF20。利用扫描电子显微镜X射线能量色散谱仪(scanning electron microscopy coupled with energy-dispersive X-ray, SEM-EDX)、动态光散射(dynamic light scattering, DLS)、X射线衍射仪(X-ray diffractometer, XRD)、拉曼光谱(Raman spectroscopy)和傅里叶变换红外光谱(Fourier transform infrared spectroscopy, FTIR)对副干酪乳酪杆菌SCFF20产生的SeNPs进行纯化、冷冻干燥和系统表征。【结果】SEM-EDX分析表明,Se是生物纳米硒颗粒的主要成分。合成的SeNPs呈球形、多分散、平均粒径约为500.62 nm。XRD图谱和拉曼光谱证实所制备纳米硒颗粒的生物无定形性质。FTIR分析证明蛋白质、胞外多糖和脂质包覆在SeNPs表面。电感耦合等离子体发射光谱(inductively coupled plasma-optical emission spectroscopy, ICP-OES)测得SeNPs的还原率为91.42%。【结论】本研究证实了副干酪乳酪杆菌SCFF20作为纳米硒生产益生菌的潜力,可作为安全生产生物源纳米硒的生物工厂以便用于营养补充剂和功能食品     

植物乳杆菌胞外多糖包覆的高稳定性硒纳米颗粒的制备及其抗氧化活性的研究*

目的:以植物乳杆菌胞外多糖(EPS)作为稳定剂和包覆剂,安全、简便地制备高稳定性胞外多糖-纳米硒复合物(E-SeNPs),并研究其稳定性和抗氧化活性。方法:将植物乳杆菌胞外多糖引入亚硒酸钠与抗坏血酸的反应体系中,室温合成E-SeNPs。采用透射电子显微镜(TEM)、动态光散射(DLS)、紫外可见光谱(UV-vis)和傅里叶变换红外光谱(FT-IR)等技术对E-SeNPs的尺寸、形貌、结构及稳定性进行研究。此外,通过检测E-SeNPs的还原能力、ABTS⁺的清除率评估其体外抗氧化活性。结果:制备了具有良好分散性、稳定性的E-SeNPs,其平均粒径为(45.17±11.9)nm,带负电荷(-31.3mV)。同时,由于包覆作用,该E-SeNPs在水溶液中可稳定存在20天。最后,相同浓度下,E-SeNPs的还原力、ABTS⁺清除率都明显高于EPS和硒纳米颗粒(SeNPs),表现出了良好的抗氧化活性。结论:获得了一种新型的SeNPs稳定剂和包覆剂,简便、安全地制备了高稳定性、水分散性良好且具有良好抗氧化活性的SeNPs。     

植物乳杆菌胞外多糖包覆的高稳定性硒纳米颗粒的制备及其抗氧化活性的研究*

目的:以植物乳杆菌胞外多糖(EPS)作为稳定剂和包覆剂,安全、简便地制备高稳定性胞外多糖-纳米硒复合物(E-SeNPs),并研究其稳定性和抗氧化活性。方法:将植物乳杆菌胞外多糖引入亚硒酸钠与抗坏血酸的反应体系中,室温合成E-SeNPs。采用透射电子显微镜(TEM)、动态光散射(DLS)、紫外可见光谱(UV-vis)和傅里叶变换红外光谱(FT-IR)等技术对E-SeNPs的尺寸、形貌、结构及稳定性进行研究。此外,通过检测E-SeNPs的还原能力、ABTS⁺的清除率评估其体外抗氧化活性。结果:制备了具有良好分散性、稳定性的E-SeNPs,其平均粒径为(45.17±11.9)nm,带负电荷(-31.3mV)。同时,由于包覆作用,该E-SeNPs在水溶液中可稳定存在20天。最后,相同浓度下,E-SeNPs的还原力、ABTS⁺清除率都明显高于EPS和硒纳米颗粒(SeNPs),表现出了良好的抗氧化活性。结论:获得了一种新型的SeNPs稳定剂和包覆剂,简便、安全地制备了高稳定性、水分散性良好且具有良好抗氧化活性的SeNPs。     

血液杆菌Haem atobacter sp.细胞脂肪酸测定的影响因素

应用美国MIDI公司Sherlock MIS系统检测一株归属于血液杆菌属的新菌种Haematobacter sp.HNMC11807的细胞脂肪酸组成,探讨不同培养方法和条件对其细胞脂肪酸组成的影响。采用不同培养条件培养微生物细胞,即不同的培养基组成、固液类型和培养时间;分析比较细胞脂肪酸检测结果存在的差异。同时对比分析仪器专用的两种检测体系,即常规标准和快速标准体系,对样品脂肪酸测定结果的影响。结果发现,细胞脂肪酸组成与其培养条件密切相关,且差异显著。因此,在应用该系统做微生物分类鉴定时,必需严格遵守特定的、统一的培养条件平行进行。     

Mariannaea sp. HJ合成纳米金银合金的特性考察

利用微生物合成纳米金银合金(Au-AgNPs)具有操作简便、生态友好等特点,但目前利用真菌合成的相关研究较少。本研究利用真菌Mariannaea sp. HJ胞内提取物合成纳米金银合金,考察了不同的金银离子浓度比例对生物合成纳米金银合金特性的影响。实验表明,金银离子浓度比例对生物纳米金银合金的组成影响较大,随着银离子浓度比例的增加,反应体系颜色会由浅紫色逐渐变为棕色,紫外-可见特征吸收峰发生了明显的蓝移,合成的纳米金银合金中银的比例也会逐渐增加。透射电子显微镜表明纳米金银合金的形貌主要为球形和伪球形,在0.5∶0.5、0.5∶1.5以及0.5∶3.0三种金银离子浓度比例下,纳米颗粒的平均粒径分别为19.24 nm、15.99 nm和19.33 nm。X射线衍射光谱结果显示纳米金银合金的晶胞为面心立方结构。利用傅里叶转换红外线光谱表征推测参与纳米金银合金还原稳定的官能团可能为-OH、-NH_3、-COOH。此外,本研究以4-硝基苯酚为底物探究生物纳米金银合金的催化特性,结果表明纳米金银合金对4-硝基苯酚具有良好的催化活性,其催化反应速率常数为7.85×10–3 s–1。上述结果表明,真菌Mariannaea sp. HJ能够合成分散性较好的纳米金银合金,在催化还原硝基芳烃污染物方面具有潜在的应用价值。     

纳米硒对肉鸡肝细胞中细胞谷胱甘肽过氧化物酶活性的影响

以亚硒酸钠和蛋氨酸硒为对照,研究了纳米单质硒(纳米硒)对肉鸡肝细胞中细胞谷胱甘肽过氧化物酶(cGPx)活性的影响。每种硒源分别以0.01、0.05、0.10、0.30、0.50、1.0μmol/L6个硒添加浓度培养肉鸡肝细胞,测定培养后0、24、48、72、96h肉鸡肝细胞cGPx活性。结果显示:亚硒酸钠添加浓度(以硒计)在0.01￣0.10μmol/L、蛋氨酸硒和纳米硒添加浓度(以硒计)在0.01￣0.30μmol/L,cGPx活性随着硒添加浓度的增加而增加;亚硒酸钠添加浓度在0.10￣1.0μmol/L、蛋氨酸硒添加浓度在0.30￣1.0μmol/L,cGPx活性随着硒添加浓度的增加而下降,而纳米硒添加浓度在0.30￣1.0μmol/L,cGPx活性始终保持在高峰平台。结果表明,3种硒源的剂量-效应关系曲线中的最适剂量范围宽度依次为:纳米硒>蛋氨酸硒>亚硒酸钠。     

乳酸菌素Gassericin T基因的合成及其在大肠杆菌中的融合表达

目的:在大肠杆菌系统中表达有抗菌活性的乳酸菌素Gassericin T。方法:根据乳酸菌素Gassericin T的基因序列,把Gassericin T的结构基因gatA编码的氨基酸的密码子转换成大肠杆菌偏爱的形式;用人工合成的寡核苷酸片段,通过重叠PCR法扩增得到gatA片段(gat基因);将合成的gat基因插入pGEX-4T-1,构建pGEX-4T-1-gat融合表达载体,转化大肠杆菌DH5α株,IPTG诱导表达,经超声裂解后获得包涵体蛋白,经溶解、变性、复性处理后获得GST-Gassericin T融合蛋白;用琼脂扩散法测定其对金黄色葡萄球菌、大肠杆菌、李斯特菌、枯草杆菌等的抗菌活性。结果与结论:采用pGEX-4T-1融合表达系统在大肠杆菌中表达了有活性的Gassericin T,融合蛋白以包涵体形式存在。复性的融合蛋白对金黄色葡萄球菌和大肠杆菌有明显的抑制作用,对李斯特菌的抑制作用不明显。     

希瓦氏菌Shewallena oneidensis MR-1合成硒纳米棒

摘要:【目的】探索采用希瓦氏菌合成硒(Se)纳米棒,并阐明合成底物Se(IV)的浓度与细菌培养时间对生物合成的影响。【方法】将希瓦氏菌Shewallena oneidensis MR-1 接种至Luria-Bertani(LB)液体培养基,分别以Se(IV)浓度0.1、1、10和100 mmol/L的Na2 SO3作为电子受体,厌氧培养并绘制生长曲线。再将希瓦氏菌接种到含最适Se( IV)浓度的LB 培养基中,在厌氧培养后第24和72 h离心获取沉淀。采用扫描电镜、X射线能谱和X射线衍射对沉淀进行分析。【结果】在Se(IV)浓度1 mmol/L的培养基中培养24 h形成的纳米棒沉淀截面直径约80 nm,长度2－3 μm。而培养72 h形成的沉淀较大,超出纳米物质范畴。采用X射线能谱和X射线衍射确定纳米棒组成为单质Se。【结论】本研究为生物合成Se纳米棒提供了一种可行的方法。希瓦氏菌最适宜在1 mmol/L Se(IV)浓度下以及在对数生长期大量合成Se纳米棒,具有潜在应用价值。     

马利亚霉菌Mariannaea sp. HJ合成TeNPs及其抗菌和抗氧化性能

【背景】近年来,纳米碲(tellurium nanoparticles,TeNPs)在光电、能源和医学等领域的应用增加。微生物合成TeNPs具有绿色低毒和条件温和等优势,受到了广泛关注,然而目前关于真菌合成TeNPs的研究较少。【目的】探究真菌Mariannaea sp. HJ合成TeNPs的能力及其抗菌、抗氧化性能。【方法】利用真菌Mariannaea sp. HJ合成TeNPs并对其合成条件进行优化,采用扫描电子显微镜、Zeta电位分析及X射线衍射仪对TeNPs进行表征。此外,通过抗菌和抗氧化实验探究TeNPs的应用。【结果】菌株HJ在菌体湿重为1.5 g及TeO32-浓度为5 mmol/L的条件下还原率最高,合成的TeNPs主要为球形;X射线衍射仪表明TeNPs的结构为六方晶系;FTIR表明羟基、羧基和氨基等官能团可能参与了TeNPs的合成。抗菌实验表明,TeNPs对金黄色葡萄球菌(Staphylococcus aureus)具有良好的抗菌性;抗氧化实验表明,TeNPs对DPPH自由基具有抑制作用,当TeNPs浓度为500mg/L时抑制率可达80%。【结论】本实验提供了一种TeN...     

利用Mariannaea sp.HJ菌株胞内提取物合成纳米银及其抗菌特性研究

【目的】近年来,纳米银由于其自身独特的抗菌活性而受到越来越多的关注,有研究表明纳米银是一种广谱的抗菌剂,其对数十种致病微生物都有强烈的抑制和杀灭作用。相较于传统的合成方法而言,具有反应条件温和、环境友好等优势的生物合成法是目前的研究热点。【方法】本研究利用真菌Mariannaea sp. HJ的胞内提取物合成纳米银,并对其合成条件进行优化调控,还进一步考察了合成的纳米银颗粒的抗菌性能。【结果】胞内提取物浓度350 mg/L、AgNO_3浓度5 mmol/L、pH 7.0为菌株HJ胞内提取物合成纳米银的最优条件;TEM图像表明合成的纳米银颗粒主要为球形和伪球形,分散性良好,无明显的团聚现象;XRD表明合成的纳米银晶体结构为面心立方体结构;通过FTIR分析结果推测提取物中的羟基、羧基等官能团可能参与了纳米银的还原和稳定过程。此外,在本实验条件下合成的纳米银颗粒对革兰氏阴性菌Escherichia coli BL21和革兰氏阳性菌Arthrobacter sp. W1都有较好的抗菌活性。【结论】真菌Mariannaea sp. HJ胞内提取物能合成尺寸均一且分散性良好的球形纳米银颗粒,合成的纳米银颗粒在抗菌方面具有潜在的研究价值。     

Antioxidant and cytotoxic effect of biologically synthesized selenium nanoparticles in comparison to selenium dioxide

The present study was designed to evaluate antioxidant and cytotoxic effect of selenium nanoparticles (Se NPs) biosynthesized by a newly isolated marine bacterial strain Bacillus sp. MSh-1. An organic–aqueous partitioning system was applied for purification of the biogenic Se NPs and the purified Se NPs were then investigated for antioxidant activity using DPPH scavenging activity and reducing power assay. Cytotoxic effect of the biogenic Se NPs and selenium dioxide (SeO₂) on MCF-7 cell line was assesed by MTT assay. Tranmission electron micrograph (TEM) of the purified Se NPs showed individual and spherical nanostructure in size range of about 80–220 nm. The obtained results showed that, at the same concentration of 200 μg/mL, Se NPs and SeO₂ represented scavenging activity of 23.1 ± 3.4% and 13.2 ± 3.1%, respectively. However, the data obtained from reducing power assay revealed higher electron-donating activity of SeO₂ compared to Se NPs. Higher IC₅₀ of the Se NPs (41.5 ± 0.9 μg/mL) compared to SeO₂ (6.7 ± 0.8 μg/mL) confirmed lower cytotoxicity of the biogenic Se NPs on MCF-7 cell line.     

Biomineralization Abilities of Cupriavidus Strain and Bacillus subtilis Strains In Vitro Isolated from Speleothems,Rani Cave,Chhattisgarh, India

In vitro culture experiments using three bacterial strains CSJC1, CSJC2, and CSJC3 isolated from speleothems, Rani cave, Chhattisgarh, India, were studied to examine their biomineralization potential. These speleothems showed high microbial cell enumerations on nutrient agar and iron agar (9 × 10⁴ CFU/g) followed by thiosulfate agar (7 × 10⁴ CFU/g), and 60 diverse strains were isolated. The BLASTn sequence search of 16S rRNA sequences with the NCBI database to establish the identity of CSJC1, CSJC2, and CSJC3 strains yielded similarity scores of ≥99% with the respective organisms, and the strains were identified as CSJC1 – Bacillus sp., CSJC2 – Cupriavidus sp., CSJC3 – Bacillus sp. The phylogenetic analysis of CSJC2 strain suggests that it formed a separate major cluster with Cupriavidus sp. and Cupriavidus necator. The phylogenetic analysis of CSJC1 and CSJC3 strains revealed that it formed a major cluster with several strains of Bacillus sp. and Bacillus subtilis. The biominerals induced by Cupriavidus sp. CSJC2 strain imaged with an ultra high-resolution field emission scanning electron microscope (FE-SEM) were seen as calcified coccoid shells that transformed into calcified dumbbells. FE-SEM imaging of biominerals induced by B. subtilis CSJC1 and CSJC3 tested both on B4 media and sheep blood agar individually showed that the precipitates formed calcified dumbbells that were almost similar but not identical phenotypically, indicating that strain-specific morphologies and crystal formation is easier when Ca is present in the media. This is the first comprehensive report on the possible evidences about the role of Cupriavidus sp. in calcite precipitation isolated from speleothems in the Indian caves. These results allow us to postulate that the identified strains may have a role in the biogenic influences in mineral formations at Rani cave.     

Enhanced production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer and antimicrobial yellow pigmentation from Cupriavidus sp. USMAHM13 with antibiofilm capability

Antibiofilm polymers have the ability to inhibit bacterial biofilm formation, which is known to occur ubiquitously in the environment and pose risks of infection. In this study, production of P(3HB-co-4HB) copolymer and antimicrobial yellow pigment from Cupriavidus sp. USMAHM13 are enhanced through medium optimization. Before the improvement of yellow pigment production, screening for the best additional supplement was performed resulting in high-yield yellow pigmentation using yeast extract with optimum concentration of 2?g/L. Effects of different concentrations of 1,4-butanediol, ammonium acetate, and yeast extract were studied using central composite design. Under optimal conditions, 53?wt% of polyhydroxyalkanoate (PHA) content, 0.35?g/L of pigment concentration, and 5.87?g/L of residual biomass were achieved at 0.56?wt% C of 1,4-butanediol, 1.14?g/L of ammonium acetate, and 2?g/L of yeast extract. Antibiofilm tests revealed that the yellow pigment coated on P(3HB-co-4HB) copolymer had significant effect on the inhibition of bacteria proliferation and colonization from 6?hr onward reaching 100% inhibition by 12?hr, hence effectively inhibiting the biofilm formation.     

CeO2 nanoparticles synthesized through green chemistry are biocompatible: In vitro and in vivo assessment

Widespread use of cerium oxide (CeO₂) nanoparticles (NPs) is found in almost all areas of research due to their distinctive properties. CeO₂ NPs synthesized via green chemistry have been characterized for antioxidant, phytochemical, and biological potential. Physical characterization through scanning electron microscopy, XRD, and TGA showed that the NPs are circular in shape, 20‐25 nm in size, and stable in a wide range of temperature. NPs display significant antioxidant (32.7% free radical scavenging activity) and antileishmanial (IC₅₀ 48 µg mL^?1) properties. In vitro toxicity tested against lymphocytes verified that NPs are biocompatible (99.38% viability of lymphocytes at 2.5 μg mL^?1). In vivo toxicity experiments showed no harmful effects on rat serum chemistry and histology of various organs and did not even change the concentration of antioxidative enzymes, total protein contents, lipid peroxidation, and nitrosative stress. These observations are in line with the statement that plant‐based synthesis of CeO₂ NPs lessens or nullifies in vitro and in vivo toxicity and hence CeO₂ NPs are regarded as a safe and biocompatible material to be used in drug delivery.     

Biochemical characterization of an alcohol dehydrogenase from Ralstonia sp.

Stereoselective reduction towards pharmaceutically potent products with multi‐chiral centers is an ongoing hot topic, but up to now catalysts for reductions of bulky aromatic substrates are rare. The NADPH‐dependent alcohol dehydrogenase from Ralstonia sp. (RADH) is an exception as it prefers sterically demanding substrates. Recent studies with this enzyme indicated outstanding potential for the reduction of various alpha‐hydroxy ketones, but were performed with crude cell extract, which hampered its detailed characterization. We have established a procedure for the purification and storage of RADH and found a significantly stabilizing effect by addition of CaCl₂. Detailed analysis of the pH‐dependent activity and stability yielded a broad pH‐optimum (pH 6–9.5) for the reduction reaction and a sharp optimum of pH 10–11.5 for the oxidation reaction. The enzyme exhibits highest stability at pH 5.5–8 and 8–15°C; nevertheless, biotransformations can also be carried out at 25°C (half‐life 80 h). Under optimized reaction parameters a thorough study of the substrate range of RADH including the reduction of different aldehydes and ketones and the oxidation of a broad range of alcohols was conducted. In contrast to most other known alcohol dehydrogenases, RADH clearly prefers aromatic and cyclic aliphatic compounds, which makes this enzyme unique for conversion of space demanding substrates. Further, reductions are catalyzed with extremely high stereoselectivity (>99% enantio‐ and diastereomeric excess). In order to identify appropriate substrate and cofactor concentrations for biotransformations, kinetic parameters were determined for NADP(H) and selected substrates. Among these, we studied the reduction of both enantiomers of 2‐hydroxypropiophenone in more detail. Biotechnol. Bioeng. 2013; 110: 1838–1848. © 2013 Wiley Periodicals, Inc.     

产碱杆菌NX-3胞外多糖的结构与性能

一株产碱杆菌NX-3分泌的可溶性胞外多糖（PS-238）经醇析、Sevag法脱蛋白、透析、冷冻干燥得分析样品,酸水解后经TLC、GC以及化学法、IR和高碘酸氧化分析测得PS-238是由葡萄糖、甘露糖、鼠李糖和葡萄糖醛酸组成的杂多糖,糖苷键类型主要为（1→3）和（1→4）。此外考察多糖溶液粘度与剪切速率、溶液浓度、温度、酸碱度以及盐离子种类和浓度等因素之间的关系,结果表明多糖水溶液的流体特征表现为假塑型,25℃时1％的水溶液粘度可达3,300cp,耐酸碱,在pH2～13的范围内粘度稳定;耐高温,在150℃内不发生明显降解;部分盐类对PS-238的增稠性有协同效应。