Schwertmannite formation adjacent to bacterial cells in a mine water treatment plant and in pure cultures of Ferrovum myxofaciens

Schwertmannite has previously been found in iron- and sulfate-rich mine waters at pH 2.8-4.5. In the present study, schwertmannite (Fe(8)O(8)(OH)(6)SO(4)) was shown to be the major mineral in a mine water treatment plant at pH 3, in which ferrous iron is mainly oxidized by bacteria belonging to the species Ferrovum myxofaciens. Strain EHS6, which is closely related to the type strain of Fv. myxofaciens, was isolated from the pilot plant and characterized as an acidophilic, iron-oxidizing bacterium. In contrast to the pilot plant, the mineral phase formed by a pure culture of Fv. myxofaciens EHS6 was a mixture of schwertmannite and jarosite (KFe(3)(SO(4))(2)(OH)(6)). In contrast to other reports of neutrophilic, iron-oxidizing bacteria, acidophilic microorganisms in the pilot plant and cultures of strain EHS6 did not show encrustation of the cell surface or deposition of minerals inside the cell, though a few cells appeared to be in contact with jarosite crystals. It was concluded that no direct biomineralization occurred in the pilot plant or in laboratory cultures. The lack of encrustation of bacterial cells in the pilot plant is considered advantageous since the cells are still able to get in contact with ferrous iron and the iron oxidation process in the mine water treatment plant can proceed.     

3 种脂环酸芽孢杆菌分离培养基的性能评价

目的：比较3 个厂家的K培养基、酵母粉淀粉葡萄糖（yeast extract starch glucose medium,YSG）培养基和耐酸耐热芽孢菌（Alicyclobacillus）培养基（Bacillus acidoterrestris medium,BAT）对脂环酸芽孢杆菌的检测效果。方法：选用标准菌株、分离株及实际样品对3 个厂家（广东环凯微生物科技有限公司（简称HK,下同）、厂家Ⅰ和厂家Ⅱ）的K培养基、YSG培养基和BAT培养基进行生长菌落数及分离效果的评估。结果：酸土脂环酸芽孢杆菌标准株和脂环酸芽孢杆菌分离株4-2在K、YSG和BAT 3 种培养基上生长的菌落数（3 个厂家培养基上的菌落数平均数）均无显著性差异（P＞0.05）;酸热脂环酸芽孢杆菌标准株和脂环酸芽孢杆菌分离株5-3在K培养基上不生长,在YSG培养基和BAT培养基上生长良好,其菌落平均数统计学上无显著性差异（P＞0.05）;对实际样品中酸土脂环酸芽孢杆菌的分离,3 个厂家的同种培养基分离率和符合率相当;对实际样品中酸热脂环酸芽孢杆菌的分离,在YSG和BAT培养基上,HK和厂家Ⅱ的分离率和符合率相当,优于厂家Ⅰ。结论：K培养基不适合所有脂环芽孢杆菌,但分离酸土脂环酸芽孢杆菌效果很好,YSG培养基和BAT培养基适合所有脂环酸芽孢杆菌。HK和厂家Ⅱ的培养基优于厂家Ⅰ。     

Optimization of medium composition for the production of a probiotic microorganism, Lactobacillus rhamnosus, using response surface methodology

This study was undertaken to optimize yeast extract, glucose, and vitamin concentrations; and also culture pH for maximizing the growth of a probiotic bacterium, Lactobacillus rhamnosus, and to assess the effects of these factors by using response surface methodology. A central composite design was used as an experimental design for the allocation of treatment combinations. A polynomial regression model with cubic and quartic terms was used for analysis of the experimental data. It was found that the effects involving yeast extract, glucose, vitamins and pH on the growth of L. rhamnosus were significant, and the strongest effect was given by the yeast extract concentration. Estimated optimum conditions of the factors for the growth of L. rhamnosus are as follows: pH=6.9; vitamin solution=1.28% (v/v); glucose=5.01% (w/v) and yeast extract=6.0% (w/v).     

Optimization of Medium Composition for Cultivating Clostridium butyricum with Response Surface Methodology

ABSTRACT: Probiotic bacteria, such as some strains of Clostridium butyricum , have been frequently used as the active ingredient in functional foods. However, slow growth of the probiotic bacteria has been one of the big concerns for their potential commercial application. In the present study, a fractional factorial design was applied to investigate the main factors, namely, the concentrations of the glucose, tryptone, yeast extract, beef extract, cys-teine, (NH₄) ₂SO₄, NaCl, K₂HPO₄, and the medium initial pH that affected the growth of 1 probiotic strain, C. butyricum ZJUCB, currently preserved in our laboratory. Central composite experimental design and response surface methodology were adopted to derive a statistical model for optimizing the composition of the medium. The experimental results showed that the optimum medium for incubating the C. butyricum ZJUCB was composed of 2.44% (w/v) glucose, 2.08% yeast extract (w/v), 1% tryptone (w/v), 0.1% (NH₄)₂SO₄ (w/v), 0.1% NaHCO₃ (w/v), 0.02% MnSO₄. H₂O (w/v), 0.02% MgSO₄. 7H₂O (w/v), 0.002% CaCl₂ (w/v), and 2% agar (w/v) (if necessary) at pH 8.55. After incubation for 24 h in the optimum medium, the populations of the viable organisms could reach 10⁹ colony-forming units (CFU)/mL, which was 100 times higher than that incubated in the initial medium.     

呕吐毒素脱毒菌Devosia sp.发酵培养基优化

德沃斯氏菌(Devosia sp.)是一种可以高效脱除谷物及饲料中呕吐毒素的新型微生物菌株,其转化产物是目前研究较为安全的呕吐毒素转化产物。本研究以碳、氮源单因素实验为基础,选用8个因素进行部分因子试验(FFD),筛选出酵母浸粉、酵母膏和蔗糖3个最显著影响因素;对3个最显著影响因素进行最陡爬坡实验获得其最优点区域;再通过中心组合实验设计(CCD)进行响应面分析得到了Devosia sp.的最适发酵培养基组成为:酵母浸粉10.00 g/L,酵母膏6.49 g/L,蔗糖8.40 g/L,硫酸铵2 g/L,磷酸二氢钾2 g/L,硫酸镁0.7 g/L,氯化钙0.02 g/L和硫酸亚铁0.02 g/L。使用该培养基进行发酵验证试验,培养40 h的OD₆₀₀达到21.66,与预测值(21.62)基本相符。综上,该回归模型对Devosia sp.发酵培养基的优化是可行的,且优化后较原始培养基生物量(OD₆₀₀)提高了89.1%,降低了生产成本,为呕吐毒素脱毒菌株的工业生产奠定了基础。     

Thermally stable hydrogels from enzymatically oxidized polysaccharides

Polysaccharides guar galactomannan (guar gum), locust bean galactomannan (locust bean gum) and tamarind galactoxyloglucan were selectively oxidized by galactose oxidase. The degrees of oxidation of the products were 18-28% for guar galactomannan, 10-16% for locust bean galactomannan and 12-14% for tamarind galactoxyloglucan, calculated from the ratio of oxidized galactose units and total carbohydrates. The rheological properties of the unoxidized and oxidized polysaccharide solutions were investigated by determining their viscosities, storage and loss moduli, and temperature dependence of moduli from 20 °C to 90 °C. All the studied oxidized polysaccharides formed hydrogels throughout the entire temperature range. Concentration (0.2-1% w/v) and degree of oxidation had an effect on the gel formation. The oxidized galactomannans formed stable gels already in low concentrations, such as 0.2-0.4% w/v, while oxidized galactoxyloglucan required a concentration of 0.8% w/v to be stable up to 90 °C. The oxidized polysaccharide hydrogels are highly potential materials for food and medical applications requiring thermal stability.     

Optimization of Medium Composition for Nisin Fermentation with Response Surface Methodology

ABSTRACT:  Nisin is an effective food biopreservative widely used in food industry. However, 1 problem of concern is limited production rate and final nisin concentration. A nisin-producing strain, L. lactis Lac2, a mutant strain with high yield of nisin, was obtained in our laboratory recently. In the present study, a fractional factorial design was applied to investigate the main factors that affect the yield of L. lactis Lac2. Central composite experimental design and response surface methodology were adopted to derive a statistical model for optimizing the composition of the medium. The results showed that the optimum medium for nisin production of L. lactis Lac2 was composed of 2.68% sucrose (w/v), 0.5% tryptone (w/v), 1% yeast extract (w/v), 0.3% Tween-80 (w/v), 0.02% MgSO₄·7H₂O (w/v), 0.81% NaCl (w/v), 1.91% K₂HPO₄ (w/v), 0.05% ascorbic acid (w/v), and 2% agar (w/v) (if necessary) at pH 6.5. When cultured in the optimum medium, the nisin yield is an average of 3381.81 IU/mL, which nearly doubled the yield when incubated in the initial medium. Also, the concentration of tryptone was decreased while that of the sucrose was increased when compared with CM broth, which means a reduction of the fermentation cost.     

Denitrification activity of the bacterium Pseudomonas sp. ASM-2-3 isolated from the Ariake Sea tideland

A new denitrifying bacterium strain ASM-2-3 was isolated from the Ariake Sea tideland, Japan. The isolate had the capability to fully remove as high as 225.8 mg nitrate-nitrogen.l(-1) under stationary culture conditions without accumulation of nitrite as an intermediate. From biochemical tests and 16S rDNA sequencing analysis, the genus of the bacterium was identified as Pseudomonas and close to stutzeri species. The nitrate removal efficiency of the isolate was faster than that of the control strain Pseudomonas stutzeri NBRC 14165, using succinate as the sole carbon source. The isolate could grow in up to 10% (w/v) of NaCl containing medium. The enzymatic tests showed that the activity of enzymes responsible for the reduction of nitrate and nitrite in strain ASM-2-3 was 1.4 and 2.3 times higher than that of the control strain. The feasibility of application of the isolate strain ASM-2-3 in a packed bed bioreactor was investigated for 40 d.     

酱油增香用耐盐酵母的分离及生长特性分析

从酱醪中分离筛选出2株耐盐产香酵母菌C5、C6,并对其耐盐性及牛长特性进行研究。试验结果表明,2株酵母菌均能耐受24％（w／w）NaCl浓度,其在低盐稀态发酵生酱油的最适生长条件为温度28℃～32℃,pH4．5～5．5,酒精含量O～1．2％vol,葡萄糖或果糖添加量2％-4％（w／w）。此外,结果显示,菌株C5的还原糖消耗能力、酒精耐受性和产酒精能力均强于C6。     

Natural antioxidant extract from galangal (Alpinia galanga) for minced beef

Raw and cooked minced beef samples containing commercial antioxidants or galangal (Alpinia galanga) extract were evaluated for storage stability at 4 ± 1 °C. Thiobarbituric acid (TBA) values of raw and cooked samples containing galangal extract at 5 and 10% (w/w) were significantly (p < 0.05) lower than those of the controls. In raw beef, galangal extract at 10% (w/w) was as effective as α‐tocopherol at 0.10% (w/w) and butylated hydroxytoluene (BHT) at 0.02% (w/w) in inhibiting/minimising lipid oxidation. Galangal extract also delayed the induction period of lipid oxidation in cooked beef but was again observed to be effective at a higher concentration than the commercial antioxidants studied. No difference in total plate count was observed in raw beef, but samples with galangal extract had higher Hunter a (redness) and chroma (colourfulness relative to the brightness of the surroundings) values. However, cooked beef treated with galangal extract or antioxidants had lower Hunter a values and total microbial counts than the control. Thus galangal extract may be a possible natural antioxidant source for meat and meat products. © 2000 Society of Chemical Industry     

Isolation of iron-oxidizing bacteria from corroded concretes of sewage treatment plants

Thirty-six strains of iron-oxidizing bacteria were isolated from corroded concrete samples obtained at eight sewage treatment plants in Japan. All of the strains isolated grew autotrophically in ferrous sulfate (3.0%), elemental sulfur (1.0%) and FeS (1.0%) media (pH 1.5). Washed intact cells of the 36 isolates had activities to oxidize both ferrous iron and elemental sulfur. Strain SNA-5, a representative of the isolated strains, was a gram-negative, rod-shaped bacterium (0.5-0.6x0.9-1.5 microm). The mean G+C content of its DNA was 55.9 mol%. The pH and temperature optima for growth were 1.5 and 30 degrees C, and the bacterium had activity to assimilate 14CO2 into the cells when ferrous iron or elemental sulfur was used as a sole source of energy. These results suggest that SNA-5 is Thiobacillus ferrooxidans strain. The pHs and numbers of iron-oxidizing bacteria in corroded concrete samples obtained by boring to depths of 0-1, 1-3, and 3-5 cm below the concrete surface were respectively 1.4, 1.7, and 2.0, and 1.2 x 10(8), 5 x 10(7), and 5 x 10(6) cells/g concrete. The degree of corrosion in the sample obtained nearest to the surface was more severe than in the deeper samples. The findings indicated that the levels of acidification and corrosion of the concrete structure corresponded with the number of iron-oxidizing bacteria in a concrete sample. Sulfuric acid produced by the chemolithoautotrophic sulfur-oxidizing bacterium Thiobacillus thiooxidansis known to induce concrete corrosion. Since not only T. thiooxidans but also T. ferrooxidans can oxidize reduced sulfur compounds and produce sulfuric acid, the results strongly suggest that T. ferrooxidans as well as T. thiooxidans is involved in concrete corrosion.     

Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions

The photosynthetic bacterium Rhodobacter sphaeroides accumulates 5-aminolevulinic acid (ALA), which is a precursor in tetrapyrrole biosynthesis, under light illumination and upon addition of levulinic acid as an inhibitor of ALA dehydratase. To generate an industrial strain which produces ALA in the absence of light, we sequentially mutated R. sphaeroides CR-286 using N-methyl-N'-nitro-N-nitrosoguanidine (NTG). The mutant strains were screened by cultivating in the absence of light and assayed for ALA by the Ehrlich reaction in a 96-well microtiter plate. The mutant strain CR-386, derived from R. sphaeroides CR-286, was selected as a mutant that exhibited significant ALA accumulation. While CR-286 required light illumination for ALA production, CR-386 was able to accumulate 1.5 mM ALA in the presence of 50 mM glucose, 60 mM glycine, 15 mM levulinic acid and 1.0% (w/v) yeast extract under conditions of agitation in the absence of light. The mutant strain CR-450, derived from strain CR-386, was selected further as a mutant that exhibited significant ALA accumulation but no accumulation of aminoacetone, analogue of ALA. CR-450 accumulated 3.8 mM ALA under the same conditions. In the presence of 50 mM glucose, 60 mM glycine, 5 mM levulinic acid and 1.0% (w/v) yeast extract, the mutant strain CR-520, derived from strain CR-450, and strain CR-606, derived from strain CR-520, accumulated 8.1 mM and 11.2 mM ALA, respectively. In batch fermentation, the strain CR-606 accumulated 20 mM ALA over 18 h after the addition of glycine, levulinic acid, glucose and yeast extract.     

Evaluation of direct plating methods to enumerate Alicyclobacillus in beverages

Ten agar media were evaluated for their suitability to support spore germination and colony development by six strains of Alicyclobacillus acidoterrestris, three strains of Alicyclobacillus acidocaldarius, and one strain of Alicyclobacillus cycloheptanicus. The influence of plating method (pour versus spread), incubation temperature (43 degrees C and 50 degrees C), and incubation time (up to 10 days) on colony development were determined. K agar, Alicyclobacillus medium (ALI agar), and Bacillus acidoterrestris thermophilic (BAT) agar recovered the highest numbers of spores. Orange serum agar and Hiraishi glucose yeast extract agar were the least suitable. Overall, surface plating was superior to pour plating and, with the exception of one strain of A. acidocaldarius which grew better at 50 degrees C, incubation of K agar, ALI agar, and BAT agar plates at 43 degrees C or 50 degrees C resulted in recovery of equivalent numbers of spores. Essentially all viable spores were detected on media incubated for 3 days at 43 degrees C. The ability of one strain of each Alicyclobacillus species to grow in ten non-carbonated commercially manufactured beverages at 30 degrees C and 43 degrees C was markedly affected by the composition of the beverages. Results show that surface plating samples on BAT agar, followed by incubating plates at 43 degrees C for 3 days provide the most suitable conditions to enumerate ten strains of three species of Alicyclobacillus most commonly responsible for spoilage of beverages.     

Effectiveness of ethanolic galangal extract (Alpinia galanga Linn.) on inhibition of lipid oxidation in fish muscle systems

Effectiveness of ethanolic galangal extract as an antioxidant in fish mince and fillet was investigated. The extract (5.8 μm gallic acid equivalent; GAE) reduced deoxygenation of sea bass haemoglobin (Lates calcarifer) at pH 6.5. The lag time for TBARs development in washed sea bass mince mediated by the haemoglobin at pH 6.5 was extended from 2 to 8 days by added galangal extract (0.02% w/w GAE). Washing the sea bass fillet with low preformed lipid hydroperoxide (LHP) content by galangal extract solution (0.2% w/v GAE) effectively inhibited the TBARs formation throughout the storage. However, in the longtail tuna fillet (Thunnus tonggol) having a significant content of preformed LHP, there was nonsignificant difference in the reaction lag time of the samples washed with the solution and with water. Thus, galangal extract can be used as an antioxidant in fish if its application is performed before initiation of lipid oxidation.     

Solubility and Foaming Properties of Phytate-Reduced Soy Protein Isolate

A pilot scale ion exchange process was developed to produce a 75 - 77% phytate-reduced soy protein isolate. The solubility and foaming properties of this isolate were compared to those of control and commercial soy protein isolates as a function of protein concentration (5 and 10%, w/v), pH (3, 6 and 9) and preheat temperature (25, 60 and 80°C). Phytate-reduced soy protein extract exhibited minimum solubility at pH 4.8 - 5.0, compared to 4.2 - 4.5 for control soy extract. Phytate-reduced soy protein isolate was most soluble and functional at pH values below its isoelectric point (pH 3), whereas control and commercial soy isolates were generally most soluble and functional at pH values above their isoelectric point (pH 6 and 9).     

南极磷虾油醇提工艺优化及其虾青素热稳定性研究

通过单因素实验和响应面分析法对南极磷虾油的醇提工艺进行优化研究,系统考察了提取溶剂、液料比、提取时间对南极磷虾油得率、磷脂和总虾青素含量的影响。利用HPLC、GC/MS等方法对最佳醇提工艺条件制备的南极磷虾油进行主要功效成分分析,并经热处理实验,探究不同抗氧化剂对南极磷虾油中虾青素稳定性的影响。结果表明,南极磷虾油最佳醇提工艺条件为:提取溶剂为95%的乙醇、液料比为10 mL/g、提取时间为137.3 min。该工艺条件制备的南极磷虾油得率为13.6%±1.2%,磷脂和总虾青素含量分别为33.62%±2.48%、210.46±5.95 mg/kg,其中多不饱和脂肪酸含量约占总脂肪酸的41.58%,且富含维生素E和维生素A。通过添加抗氧化剂可显著提高南极磷虾油中虾青素的耐热稳定性,其作用效果由大到小依次为迷迭香提取物（0.02%,w/v）+生育酚（0.02%,w/v）、特丁基对苯二酚(TBHQ,0.02%,w/v)、迷迭香提取物（0.02%,w/v）和生育酚(0.02%,w/v)。本研究结果可为南极磷虾油的工业化生产及其贮藏稳定性提供科学依据。     

Temperature and NaCl Affect Growth and Survival of Escherichia coli 0157:H7 in Poultry-Based and Laboratory Media

In tryptic soy broth (TSB) and a poultry extract broth (PB) with 0 to 10% (w/v) NaCl incubated at 37°C, growth of E. coli 0157:H7 was inhibited at 28% NaCl whereas at 10°C, growth was inhibited at 24% NaCl in TSB and at 26% NaCl in PB. The bacterium did not grow at 4°C. Increased NaCl-sensitivity observed at 10°C was a bacteriostatic effect that was ineffective with increasing incubation temperature. At 10°C, E. coli 0157:H7 was more salt-tolerant in PB than in TSB, although PB growth rates were lower. Findings suggest that PB may be a more suitable medium for testing E. coli 0157:H7 in poultry products. Cells of E. coli 0157:H7 that were exposed to refrigeration (4°C) and/or NaCl for 24 days did not grow on MacConkey agar with 1% sorbitol.     

牛肝菌胞外多糖发酵培养基的优化

在Plackett Burman设计实验结果基础上 ,采用响应曲面法对影响牛肝菌 (Boletussp .)ACCC 5 0 3 2 8发酵胞外多糖的培养基 5个关键组成成分酵母膏 (X1)、麦芽糖 (X2 )、(NH4 ) 2 SO4(X3)、FeSO4 (X4 )和CuSO4 ·5H2 O(X5)的最佳水平范围进行了研究和探讨。通过对二次多项回归方程求解得知 ,在上述自变量取值分别为 :酵母膏 1 4 .2 g/L ,麦芽糖 2 2 .2g/L ,(NH4 ) 2 SO4 2 .7g/L ,FeSO4 66.9mg/L ,CuSO4 1 0 1 .5 9μg/L时 ,胞外多糖最大预测值为 75 2 .0 79μg/mL(发酵醪 ) ,此预测可信度不仅被统计分析所验证 ,也被实践所证实     

Thermal inactivation of Alicyclobacillus acidoterrestris spores under different temperature, soluble solids and pH conditions for the design of fruit processes

Alicyclobacillus acidoterrestris, a thermoacidophilic, non-pathogenic and spore-forming bacterium has been detected in several spoiled commercial pasteurised fruit juices. A. acidoterrestris spores, besides being resistant to the pasteurisation treatment conditions normally applied to acidic fruit products, can germinate and grow causing spoilage. Therefore, this microorganism was suggested as the target to be used in the design of adequate pasteurisation processes. The objectives of this work were to investigate the influence of temperature (T: 85-97 degrees C), total soluble solids (SS: 5-60 degrees Brix or % by weight) and pH (2.5-6.0) on D-values (decimal reduction time) of Alicyclobacillus acidoterrestris (type strain, NCIMB 13137) spores, and to fit a model using response surface methodology. A central composite face-centred experimental design was used, and the response, D-value determined in malt extract broth, ranged between 0.498+/-0.045 and 94.9+/-6.7 min. Within the factor ranges studied, temperature was the parameter that most affected the D-value. Following this was the SS and, lastly, the pH value. A linear decrease in D-value was observed with decreasing SS and pH, and a non-linear decrease in D-value was noticed with increasing temperature. A second order polynomial was successfully fitted to the data (R2 = 0.98). In general, D-values measured in real fruit systems, such as orange, apple and grape juices, blackcurrant concentrates, cupua?u (exotic fruit) extract and orange juice drink, were higher than those predicted by the malt extract broth model. This result emphasises the importance of experimental validation of any model-derived process.     

忧遁草茎中黄酮类化合物的提取工艺优化及比较

为筛选适合忧遁草茎中黄酮类化合物的提取方法,以得率为评价指标,在单因素基础上,通过正交试验优化纤维素酶法和有机溶剂浸提法提取忧遁草茎中的黄酮类化合物工艺;采用ABTS自由基法、DPPH自由基法和还原力法比较提取物的抗氧化性。结果表明,纤维素酶辅助提取忧遁草茎中黄酮类化合物的最佳工艺条件为:乙醇浓度80%(v/v)、温度55 ℃、pH5.0、底物质量浓度30 g/L、酶用量300 U/g、时间1.5 h;有机溶剂浸提的最佳工艺条件为:乙醇浓度80%(v/v)、温度55 ℃、料液比1:15 (g/mL)、时间2.5 h。纤维素酶辅助提取忧遁草茎中的黄酮类化合物得率为(1.32±0.11)%(w/w),高于有机溶剂浸提法的(1.05±0.08)%(w/w)。2种提取方法得到的黄酮化合物均具有较强的抗氧化性,其中纤维素酶辅助提取物清除ABTS自由基和DPPH自由基的半抑制浓度(IC50)值分别为(1.94±0.04)、(0.178±0.013)mg/mL,低于有机溶剂提取物的(2.76±0.05)、(0.200±0.015)mg/mL,说明酶法提取物的抗氧化性强于有机溶剂浸提法。忧遁草茎中黄酮类化合物提取采用纤维素酶辅助提取较为适宜。