杂色曲霉素的分离、纯化及鉴定

将两株杂色曲霉菌分别接种于4kg玉米豆粉固体产毒培养基,28℃静置培养35d。以甲醇-4％ KC1(9:1)混合液及甲醇-三氯甲烷混合液浸提杂色曲霉素(sterigmatocy stin,简称ST),再用三氯甲烷萃取,经硅胶柱层析法初步纯化。在甲醇、乙腈、乙醚、丙酮等溶剂中多次重结晶,最后得到2271.6mg淡黄色针状结晶。经过熔点测定、元素分析、四种光谱学鉴定、薄层层析及高效液相色谱鉴定,确认该晶体为ST。纯度在99.9％以上。     

杂色曲霉素的分离,纯化及鉴定

将两株杂色曲霉菌分别接种于4kg玉米豆粉固体产毒培养基,28℃静置培养35d。以甲醇-4％ KC1(9:1)混合液及甲醇-三氯甲烷混合液浸提杂色曲霉素(sterigmatocy stin,简称ST),再用三氯甲烷萃取,经硅胶柱层析法初步纯化。在甲醇、乙腈、乙醚、丙酮等溶剂中多次重结晶,最后得到2271.6mg淡黄色针状结晶。经过熔点测定、元素分析、四种光谱学鉴定、薄层层析及高效液相色谱鉴定,确认该晶体为ST。纯度在99.9％以上。     

不同生境来源杂色曲霉次级代谢产物研究进展

杂色曲霉是曲霉属的常见种类，广泛分布于海洋、高等植物、红树林和节肢动物等生境中，近年研究表明杂色曲霉可产生生物碱、萜类、聚酮等结构多样的次级代谢产物，具有抗菌、抗肿瘤、抗氧化、抗病毒等活性。本文总结了2017—2022年间报道的37株杂色曲霉次级代谢产物结构及其生物活性的研究进展，涉及325个次级代谢产物，其中新化合物111个。本文揭示了杂色曲霉的次级代谢产物与生境的相关性，对从更多生境来源的杂色曲霉及其它微生物资源来挖掘新天然产物具有参考作用。     

真菌和氮素在食物链中与胃癌病因关系的探讨

作者等根据假设,即真菌与氮素通过食物链过程进入人体,由于真菌毒素或其代谢产物与胃内亚硝酸盐、胍类、胺类等合成致癌性亚硝基化合物,并在慢性胃炎的基础上逐渐发展为胃癌,在我国胃癌高,低发区进行了综合考察。本文报告考察结果,发现胃液内杂色曲霉(Aspergillus versicolor)检出率最高,高低发区对比差异显著(p<0.001)。此菌与胃液的pH值、酸度、NO_2~-含量、胃粘膜活检结果关系密切。已知杂色曲霉产的杂色曲霉素(sterigmatocystin)可诱发大鼠的腺胃癌,因此考虑此菌与人胃癌的发生可能有关,但是否为因果关系,尚需进一步实验研究。     

相似蜂海绵相关真菌杂色曲霉F62活性代谢产物研究

研究了1株相似蜂海绵相关真菌F62的活性代谢产物,经分子系统学分析表明该真菌属于杂色曲霉。将F62菌株用大米固体培养基在室温（约25℃）下静置培养45d,经乙酸乙酯超声萃取得到粗提物。通过硅胶柱层析、Sephadex LH-20凝胶柱层析和HPLC等色谱手段,分离得到了6个化合物。通过核磁共振、质谱等波谱分析手段可鉴定出其结构分别为Alantrypinone（1）、洛伐他汀（2）、甲酯型莫纳克林K（3）、土曲霉酮（4）、土震素B（5）和麦角甾醇（6）。化合物1系首次从该属真菌中分离得到,化合物2－5为首次从该种真菌中分离得到。首次对化合物3的碳谱数据进行报道。初步的药理研究表明,化合物4具有体外抗炎活性。     

应用薄层层析技术快速检出杂色曲霉素产毒菌株

直接硅胶板薄层层析快速检出杂色曲霉素产毒菌株,简化了培养、萃取纯化等繁琐步骤,大大缩短样品检测所需时间,做到快速灵敏。检验时取生长待测菌株的琼脂块,直接以硅胶薄层板展开浸出溶液,显色后产毒株色谱带中毒素斑点清晰易辨。所用试剂、设备简单,费用低廉,容易掌握。尤其适用于大量菌株的分离筛选。应用此方法,检测了222个真菌菌株的产毒能力。     

红曲霉液态发酵生产Monacolin K培养基的优化

利用正交试验设计对红曲霉TQ-57发酵生产Monacolin K的摇瓶发酵培养基进行了初步研究。结果表明:Monacolin K的最适发酵碳源为甘油,最佳添加质量分数为14%,最适氮源为蛋白胨,最佳氮源质量分数为3%,培养基初始pH为6.5,培养周期为8 d,在此条件下Monacolin K产量可达27.75 mg/L。     

多壁碳纳米管固定化生物酶修饰电极检测杂色曲霉素的初步研究

色曲霉素（ST）是一种致癌致畸的真菌毒素,已报道的检测ST的技术有TLC、HPLC、ELISA以及毒素基因的PCR检测技术。初步探索了酶生物传感器三电极系统对ST的电化学分析。在实验中,应用多壁碳纳米管作为分子识别元件ADTZ的固定化基质和传感器的电子传递体构建了Au工作电极,对ST进行CV和DPV分析,结果表明：ST在-600mＶ位置有一明显的特征还原峰电位,线性检测范围是8.32×10^-5～6656×10^-5mg/mL,检测下限为8.32×10^-5mg/mL,响应时间10s,为进一步的研究打下良好的基础。     

应用薄层层析技术快速检出杂色曲霉素产毒菌株

直接硅胶板薄层层析快速检出杂色曲霉素产毒菌株,简化了培养,萃取纯化等繁琐步骤,大大缩短样品检测所需时间,做到快速灵敏,检验时取生长待测菌株的琼脂块,直接以硅胶薄层析展开浸出溶液,显色后产毒株色谱带中毒素斑点清晰易辨,所用试剂,设备简单,费用低廉,容易掌握,尤其适用于大量菌株的分离筛选,应用此方法,检测了２２２个真菌菌株的产毒能力。     

杂色云芝产漆酶的发酵条件研究*

本文对杂色云芝（Coriolus versicolor）产漆酶的发酵条件作了研究。结果表明摇瓶实验产漆酶（Laccase）的最佳培养基成分为：可溶性淀粉 2g/L, NH4Cl 24mmol/L, 微量元素混合液 7ml/L, pH3.0柠檬酸—Na2HPO4缓冲溶液 0.01mol/L, KH2PO4 1.4×10-2 mol/L, MgSO4·7H2O 2.03×10-3mol/L, CaCl2·2H2O 6.8×10-4 mol/L, VB1 2.97×10-6 mol/L, 吐温80 4.0g/L, 愈创木酚0.01mmol/L, CuSO4 ·5H2O 0.005mmol/L,最佳发酵条件为培养基初始pH3.0, 菌体生长6d,培养基装量为250ml三角瓶中25ml培养液,25℃条件下振荡培养(150r/min)9d。     

Effect of Medium Components on Bacteriocin Production by Lactobacillus Pentosus ST151BR,a Strain Isolated from Beer Produced by the Fermentation of Maize,Barley and Soy Flour

Lactobacillus pentosus ST151BR, isolated from home-brewed beer, produces a 3.0 kDa antibacterial peptide (bacteriocin ST151BR) active against Lactobacillus casei, Lactobacillus sakei, Pseudomonas aeruginosa, Enterococcus faecalis and Escherichia coli. Treatment with Proteinase K or Pronase resulted in loss of activity. Bacteriocin levels of 6400 AU/ml were recorded in MRSbb (De Man-Rogosa-Sharpe broth without Tween 80) at pH 5.5, 6.0 and 6.5. The same growth conditions at pH 4.5 yielded only 1600 AU/ml bacteriocin. Inclusion of Tween 80 in the growth medium reduced bacteriocin production by more than 50%. Growth in the presence of tryptone or tryptone plus meat extract stimulated bacteriocin production, whereas much lower activity was recorded when the bacteria were grown in the presence of meat extract, yeast extract, tryptone plus yeast extract, meat extract plus yeast extract, or a combination of tryptone, meat extract and yeast extract. MRSbb supplemented with maltose, lactose or mannose (2.0%, w/v) yielded bacteriocin levels of 6400 AU/ml. Sucrose or fructose at these concentrations reduced the activity by 50 and 75%, respectively. Growth in the presence of 4.0%(w/v) glucose resulted in 50% activity loss. Glycerol levels as low as 0.1%(w/v) repressed bacteriocin production. Addition of cyanocobalamin, ascorbic acid, thiamine and thioctic acid (1.0 mg/l) to the growth medium did not lead to an increase in bacteriocin production. This revised version was published online in July 2006 with corrections to the Cover Date.     

固定化果胶酶澄清果汁的条件及效果

利用固定化果胶酶对四种不同果汁澄清条件及效果进行研究,结果表明,固定化果胶酶澄清四种不同果汁的效果明显,其中澄清桔汁的果胶酶重复使用20次以上,酶活力及透光率仍可维持在80%以上,其最适反应条件是:果汁浓度50%;pH 3.0~3.5;温度45~50℃;反应时间2小时;酶量每毫升果汁0.05 g固定化果胶酶;澄清时间20小时。     

Differences in Escherichia coli Culture Conditions Can Have a Large Impact on the Induction of Extreme Acid Resistance

A recently isolated Escherichia coli strain (3TF4) survived an acid shock that mimicked the low pH of the human gastric stomach (pH 2, 1 h), but this survival was highly influenced by prior growth conditions. Only 0.01% of the stationary phase cells that had been grown anaerobically in a carbonate medium (2 mg glucose and 0.25 mg yeast extract per ml, 40 mm sodium carbonate, final pH 6.5) survived the acid shock, and the survival of exponential phase cultures was even lower (0.0001%). Small amounts of Trypticase (1.5 mg/ml) increased the survival as much as 5000-fold, but cultures that were provided with higher concentrations of Trypticase (7.5 mg/ml) did not reach the stationary phase in 24 h and were more acid sensitive. Sodium acetate (50 mm) also increased acid resistance, and the increased acid shock survival was greater for the cells that had reached the stationary phase (100 versus 1000-fold, respectively). E. coli 3TF4 cultures that had been grown aerobically in Luria broth were already so acid resistant (survivals greater than 40%) that they did not respond to sodium acetate. E. coli 3TF4 cultures that were refrigerated (5°C, 7 days) were nearly as acid resistant as those that were immediately subjected to acid shock (pH 2.0, 1 h). Received: 20 December 2000/Accepted: 7 February 2001     

模拟胃肠环境对3株光合细菌存活率影响的实验研究

目的模拟胃肠环境对3株光合细菌(Photosynthetic bacteria,PSB)存活率影响进行体外实验研究。方法测定3株PSB分别在pH2.0、3.0和4.0的模拟胃液及含有或不含有0.45%牛胆盐(pH8.0)的模拟小肠液中不同时间的存活率。结果pH2.0时模拟胃液处理不同时间对3株PSB存活率有显著影响(P0.01);pH3.0时,模拟胃液处理60、90和180 min对菌株rub存活率有显著影响(P0.01),处理90、180 min对菌株Ga存活率有显著影响(P0.05),处理180 min对菌株2C存活率无显著影响(P0.05);pH4.0时模拟胃液处理180 min对3株PSB存活率无显著影响(P0.05)。模拟小肠液处理240 min对3株PSB菌株活菌数无显著影响(P0.05)。结论模拟胃液对3株PSB存活率影响具有菌株特异性和pH依赖性,而模拟小肠液对3株PSB存活率无显著影响。     

Increased production of bacteriocin ST4SA byEnterococcus mundtii ST4SA in modified corn steep liquor

Enterococcus mundtii ST4SA produces a broad-spectrum bacteriocin (bacST4SA), active against Gram-positive and Gramnegative bacteria. Growth in corn steep liquor (CSL) with a sugar content of 5.0 and 10.0 g/l yielded bacST4SA levels of 12800 AU/ml. A four-fold increase in bacST4SA production (51200 AU/ml) was recorded in CSL with a sugar content of 7.5 g/l supplemented with 6.5 g/l yeast extract (CSL-YE). Poor growth and low levels of bacST4SA production were observed when cells were grown in CSL-YE controlled at pH 5.5. Fermentation at pH 7.5 yielded 25600 AU/ml after 6 h, but the activity levels decreased to approximately 1000 AU/ml during the next 6 h. Adjustment of the culture pH from 6.5 to 5.5 after 6 h of fermentation extended bacST4SA activity (51200 AU/ml) over 8 h. Activity then decreased to 25600 AU/ml and was maintained this level for 10 h. Optimal levels of bacST4SA production (102400 AU/ml) were obtained after 6 h of fermentation in CSL-YE supplemented with 7.5 g/l glucose at the start of the fermentation. This level of production was maintained by changing the culture pH from 6.5 after 6 h of fermentation to pH 5.5. This study proved that bacST4SA could be produced at high levels in an inexpensive industrial medium byE. mundtii ST4SA.     

Partial characterisation of two bacteriocins produced byLactobacillus paracasei subsp.paracasei ST242BZ and ST284BZ and the effect of medium components on their production

The influence of medium components on production of bacteriocins ST242BZ (10.0 kDa) and ST284BZ (3.5 kDa) byLactobacillus paracasei subsp.paracasei ST242BZ and ST284BZ have been studied. Growth in MRS broth (pH of 6.5) yielded bacteriocin levels of 12800 AU/ml. Modified MRS with tryptone as the only nitrogen source, MRS supplemented with KH₂PO₄ (10–100 g/l), or MRS supplemented with thiamine increased bacteriocin ST242BZ production to 25600 AU/ml. Tryptone, combinations of tryptone, meat extract and yeast extract, or thiamine did not increase bacteriocin ST284BZ production. However, MRS supplemented with K₂HPO₄ (50–100 g/l) increased bacteriocin ST284BZ production up to 25600 AU/ml. Our results suggest that production of bacteriocins ST242BZ and ST284BZ are stimulated by potassium ions.     

Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi

The influence of growth parameters on the fermentative production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi was studied. The bacteriocin production was greatly affected by carbon and nitrogen sources. Strain A164 produced at least 4-fold greater bacteriocin in M17 broth supplemented with lactose than other carbon sources. The amount of 3% yeast extract was found to be the optimal organic nitrogen source. While the maximum biomass was obtained at 37 degrees C, the optimal temperature for the bacteriocin production was 30 degrees C. The bacteriocin production was also affected by pH of the culture broth. The optimal pH for growth and bacteriocin production was 6.0. Although the cell growth at pH 6.0 was nearly the same level at pH 5.5 and 6.5, the greater bacteriocin activity was observed at pH 6.0. Exponential growth took place only during an initial period of the cultivation, and then linear growth was observed. Linear growth rates increased from 0.160 g(DCW) x l(-1) x h(-1) to 0.245 g(DCW) x l(-1) x h(-1) with increases in lactose concentrations from 0.5 to 3.0%. Maximum biomass was also increased from 1.88 g(DCW) x l(-1) to 4.29 g(DCW) x l(-1). However, increase in lactose concentration did not prolong the active growth phase. After 20 h cultivation, cell growth stopped regardless of lactose concentration. Production of the bacteriocin showed primary metabolic kinetics. However, bacteriocin yield based on cell mass increased greatly during the late growth phase. A maximum activity of 131x10(3) AU x ml(-1) was obtained at early stationary growth phase (20 h) during the batch fermentation in M17L broth (3.0% lactose) at 30 degrees C and pH 6.0.     

Dextransucrase production using cashew apple juice as substrate: effect of phosphate and yeast extract addition

Cashew apples are considered agriculture excess in the Brazilian Northeast because cashew trees are cultivated primarily with the aim of cashew nut production. In this work, the use of cashew apple juice as a substrate for Leuconostoc mesenteroides cultivation was investigated. The effect of yeast extract and phosphate addition was evaluated using factorial planning tools. Both phosphate and yeast extract addition were significant factors for biomass growth, but had no significant effect on maximum enzyme activity. The enzyme activities found in cashew apple juice assays were at least 3.5 times higher than the activity found in the synthetic medium. Assays with pH control (pH = 6.5) were also carried out. The pH-controlled fermentation enhanced biomass growth, but decreased the enzyme activity. Crude enzyme free of cells produced using cashew apple juice was stable for 16 h at 30°C at a pH of 5.0.     

Human prostatic gastricsinogen: The precursor of seminal fluid acid proteinase

A gastricsinogen-like acid proteinase precursor has been purified by DEAE-cellulose, DEAE-Sephadex A-50, poly-l-lysine-Sepharose 4B, and N-acetyl-l-phenylalanyl-l-tyrosine-Sepharose 4B affinity chromatography from human prostates. The active enzyme hydrolyzes acid-denatured hemoglobin at pH 1.0 and 3.0, while two other active fractions only showed the pH 3.0 activity and resembled cathepsin D (EC 3.4.23.5). The pH optimum, milk-clotting activity, specificity toward synthetic substrates, inhibition by pepstatin, and molecular weight strongly suggest that the prostatic-derived enzyme is identical to seminal fluid and to gastric juice gastricsin.