角质细胞生长因子-2的发酵及纯化工艺研究

目的研究重组人角质细胞生长因子 2 (rhKGF 2 )工程菌的高密度发酵和提纯工艺。方法通过增加培养基营养成分 ,发酵过程补加葡萄糖 ,获得高产率菌体。细菌裂解液经硫酸铵沉淀、分子筛、肝素亲和色谱和离子交换色谱等方法进行纯化 ,电泳分析结果。结果每 1L发酵液可获得菌体 12 .2g ,表达率达 2 8.7%。纯化后rhKGF 2纯度达 97%。结论rhKGF 2工程菌可获得高密度发酵。纯化工艺适用于rhKGF 2的大量制备     

Enhanced Production of Biogas Using Biochar–Sulfur Composite in the Methane Fermentation Process

The methane fermentation of organic waste is one way to minimize organic waste, which accounts for 77% of the global municipal waste stream. The use of biochar as an additive for methane fermentation has been shown to increase the production potential of biogas. Sulfur waste has a potential application to synergistic recycling in a form of composites with other materials including biochar. A composite product in the form of a mixture of biochar and molten sulfur has been proposed. In this experiment, additions of the sulfur–biochar composite (SBC) were tested to improve the fermentation process. The biochar was produced from apple chips under the temperature of 500 °C. The ground biochar and sulfur (<1 mm particle size) were mixed in the proportion of 40% biochar and 60% sulfur and heated to 140 °C for sulfur melting. After cooling, the solidified composite was ground. The SBC was added in the dose rate of 10% by dry mass of prepared artificial kitchen waste. Wet anaerobic digestion was carried out in the batch reactors under a temperature of 37 °C for 21 days. As an inoculum, the digestate from Bio-Wat Sp. z. o. o., Świdnica, Poland, was used. The results showed that released biogas reached 672 mL × g_vs⁻¹, and the yield was 4% higher than in the variant without the SBC. Kinetics study indicated that the biogas production constant rate reached 0.214 d⁻¹ and was 4.4% higher than in the variant without the SBC.     

Influence of Fermentation and Drying Materials on the Contamination of Cocoa Beans by Ochratoxin A

Ochratoxin A (OTA) is a mycotoxin produced mainly by species of Aspergillus and Penicillium. Contamination of food with OTA is a major consumer health hazard. In Côte d’Ivoire, preventing OTA contamination has been the subject of extensive study. The current study was conducted to evaluate the influence of fermentation and drying materials on the OTA content in cocoa. For each test, 7000 intact cocoa pods were collected, split open to remove the beans, fermented using 1 of 3 different materials, sun-dried on 1 of 3 different platform types and stored for 30 days. A total of 22 samples were collected at each stage of post-harvesting operations. The OTA content in the extracted samples was then quantified by high-performance liquid chromatography. OTA was detected in beans at all stages of post-harvesting operations at varying levels: pod-opening (0.025 ± 0.02 mg/kg), fermentation (0.275 ± 0.2 mg/kg), drying (0.569 ± 0.015 mg/kg), and storage (0.558 ± 0.04 mg/kg). No significant relationships between the detected OTA level and the materials used in the fermentation and drying of cocoa were observed.     

Decrease of wheat flour allergenicity via lactic acid fermentation

Results of the research on the lowering of allergenicity of wheat flour fermented using homo- and heterofermentative lactic acid bacteria and mixed cultures with yeast were discussed. The gliadins’ immunoreactivity was measured using the indirect non-competitive enzyme-linked immunosorbent assay method. The biggest decrease of the immunoreactivity of gliadin fraction was observed in the case of fermentation performed with the use of mixed cultures of lactic acid bacteria and yeast. This way of wheat flour modification seems to be promising, because fermentation is a natural process causing not only lowering of the allergenicity, but also improving organoleptic and nutritional values of obtained products.     

六神曲质量标准研究

目的：建立六神曲的质量标准。方法：通过全国六神曲处方组成、药物剂量配比、发酵工艺和成品质量研究进行分析。结果：各地标准六神曲药物组成、剂量配比出入较大,发酵工艺不尽相同,成品的质量控制仅凭感官判断,质量较难控制。结论：制定统一的国家标准,确保六神曲临床疗效。     

Contribution of Colonic Fermentation and Fecal Water Toxicity to the Pathophysiology of Lactose-Intolerance

Whether or not abdominal symptoms occur in subjects with small intestinal lactose malabsorption might depend on differences in colonic fermentation. To evaluate this hypothesis, we collected fecal samples from subjects with lactose malabsorption with abdominal complaints (LM-IT, n = 11) and without abdominal complaints (LM-T, n = 8) and subjects with normal lactose digestion (NLD, n = 15). Lactose malabsorption was diagnosed using a ¹³C-lactose breath test. Colonic fermentation was characterized in fecal samples at baseline and after incubation with lactose for 3 h, 6 h and 24 h through a metabolomics approach using gas chromatography-mass spectrometry (GC-MS). Fecal water cytotoxicity was analyzed using a colorimetric assay. Fecal water cytotoxicity was not different between the three groups (Kruskall-Wallis p = 0.164). Cluster analysis of the metabolite patterns revealed separate clusters for NLD, LM-T and LM-IT samples at baseline and after 24 h incubation with lactose. Levels of 5-methyl-2-furancarboxaldehyde were significantly higher in LM-IT and LM-T compared to NLD whereas those of an unidentified aldehyde were significantly higher in LM-IT compared to LM-T and NLD. Incubation with lactose increased short chain fatty acid (SCFA) concentrations more in LM-IT and LM-T compared to NLD. In conclusion, fermentation patterns were clearly different in NLD, LM-IT and LM-T, but not related to differences in fecal water cytotoxicity.     

产纤溶酶海洋枯草芽孢菌的液体发酵条件优化

目的采用响应面法对高产纤溶酶海洋菌株Y-6-A的液体发酵条件进行优化,以进一步提高产酶量并考察该法的有效性。方法先采用单因素实验考查各因素对菌株产酶的影响,并在此基础上利用PlackettBurman设计筛选到影响菌株产酶的主要因素,接着通过最陡爬坡实验逼近酶活最高区域,最后根据Box-Behnken中心组合实验设计对各显著因子进行优化。结果最终得到影响产酶的因素有4个:可溶性淀粉、黄豆粕粉、温度和转速,最佳发酵条件为:可溶性淀粉46.1g/L,黄豆粕粉23.3g/L,温度31.2℃,转速184r·min^-1,在此发酵条件下得到的酶活为3 606.23IU/mL,经3次验证实验测得的酶活稳定在3 580.32±14.82IU/mL,较优化前提高了22.4%,此外,实验值与预测值之间的相对误差为0.63%。结论响应面的优化结果比较理想,经优化后能较好提高酶活。     

不同切碎长度和发酵方式对全株饲用谷子青贮质量的影响

本研究旨在探索不同切碎长度和发酵方式对全株饲用青贮谷子质量的影响。采用2×3双因子试验设计,切割长度分别为1.5 cm、2.5 cm和3.5 cm,发酵方式分为复合菌剂发酵和乳酸菌发酵两个类型,每处理3个重复。常温发酵40 d后取样,进行感官评定,分析基础养分,测定挥发性成分。结果表明,随着切碎长度增加,气味评分和总分总体呈降低趋势,中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)含量呈增加趋势,丁酸含量显著增加;相比乳酸菌发酵组,复合菌剂发酵组整体气味评分和乳酸含量较高,pH值和NDF、ADF含量较低。综上,不同切碎长度和发酵方式对于全株饲用谷子青贮质量均有显著影响。降低切碎长度和使用复合菌剂发酵均有助于促进纤维分解,减少丁酸产量,改善青贮感官评分。     

Extended-Spectrum β-Lactamases (ESBLs): Characterization,Epidemiology and Detection

β-lactamases of Enterobacteriaceae are the most important mechanism of resistance against β-lactam drugs. Two types of β-lactamases can confer resistance against 3rd generation cephalosporins. Chromosomally mediated β-lactamases are inducible and are not inhibited by clavulanic acid. Resistance due to these enzymes is non-transferable. The 2nd type of enzyme is plasmid-mediated β-lactamases, which are inhibited by clavulanic acid. These enzymes are more important clinically as these can be transferred between various species of Enterobacteria ceae. These enzymes are called extended-spectrum β-lactamases (ESBLs). ESBL-producing Enterobacteriaceae have been responsible for numerous outbreaks of infection throughout the world and pose challenging infection control issues. Antibacterial choice is often complicated by multi-resistance. ESBLs can confer resistance against all β-lactam drugs except carbapenems and cephamycins. Nursing home patients may be an important reservoir of ESBL-containing multiple antibiotic-resistant organisms. Use of broad-spectrum oral antibiotics and probably poor infection control practices may facilitate spread of this plasmid-mediated resistance. In addition to known populations at risk, ambulatory patients with chronic conditions represent another patient population that may harbor ESBL-producing organisms. Various methods can be used for detection of ESBLs in the laboratory. These tests include double disc diffusion test, Vitek ESBL test, E Tests, MIC Determination, genetic method, and isoelectric focusing (IEF).     

Engineering Escherichia coli for efficient conversion of glucose to pyruvate

Escherichia coli TC44, a derivative of W3110, was engineered for the production of pyruvate from glucose by combining mutations to minimize ATP yield, cell growth, and CO2 production (DeltaatpFH DeltaadhE DeltasucA) with mutations that eliminate acetate production [poxB::FRT (FLP recognition target) DeltaackA] and fermentation products (DeltafocA-pflB DeltafrdBC DeltaldhA DeltaadhE). In mineral salts medium containing glucose as the sole carbon source, strain TC44(DeltafocA-pflB DeltafrdBC DeltaldhA DeltaatpFH DeltaadhE DeltasucA poxB::FRT DeltaackA) converted glucose to pyruvate with a yield of 0.75 g of pyruvate per g of glucose (77.9% of theoretical yield; 1.2 g of pyruvate liters(-1).h(-1)). A maximum of 749 mM pyruvate was produced with excess glucose. Glycolytic flux was >50% faster for TC44 producing pyruvate than for the wild-type W3110 during fully aerobic metabolism. The tolerance of E. coli to such drastic changes in metabolic flow and energy production implies considerable elasticity in permitted pool sizes for key metabolic intermediates such as pyruvate and acetyl-CoA. In strain TC44, pyruvate yield, pyruvate titer, and the rate of pyruvate production in mineral salts medium were equivalent or better than previously reported for other biocatalysts (yeast and bacteria) requiring complex vitamin feeding strategies and complex nutrients. TC44 offers the potential to improve the economics of pyruvate production by reducing the costs of materials, product purification, and waste disposal.