Growth and viability of Lactobacillus acidophilus NRRL B-4495, Lactobacillus casei NRRL B-1922 and Lactobacillus plantarum NRRL B-4496 in milk supplemented with cysteine,ascorbic acid and tocopherols

The effect of three reducing agents (RAs), l-cysteine, ascorbic acid, and tocopherols, on the growth of Lactobacillus acidophilus, Lactobacillus casei, or Lactobacillus plantarum during milk fermentation was evaluated. pH, redox potential, and Lactobacillus counts were determined until pH ≈ 4.6. Further, the study aimed to optimise the concentration of the RAs by formulating the fermented milks with the same RA at different concentrations (0–250 mg L⁻¹ for l-cysteine or ascorbic acid and 0–15 mg L⁻¹ for tocopherols) using a Box–Behnken experimental design. After 45 days of refrigerated storage, the viability of each Lactobacillus species was maximised. We observed that the effect of RA on Lactobacillus is species dependent; ascorbic acid and tocopherols reduced the fermentation time (29%–43%), whereas l-cysteine enhanced the Lactobacillus counts (≥1 log₁₀ cfu mL⁻¹). Lactobacillus species differ in terms of oxygen sensitivity.     

海藻酸钠/明胶协同固定化假单胞菌B-3酶法合成L-半胱氨酸

目的:研究海藻酸钠/明肢协同固定化假单胞菌B-3的制备条件.以及固定化细胞酶法转化DL-2-氨基-△2-噻唑啉-4-羧酸(DL-ATC)生物合成L-半胱氨酸的工艺条件.方法:比较8种不同的细胞固定化方法,优选海藻酸钠/明胶协同固定化为假单胞菌B-3最佳的固定化方法,并对凝胶组成,增殖活化时间和菌体包埋量等条件进行优化;通过向转化液中添加表面活性剂Tween-60,N-氨甲酰-L-半胱氨酸酰胺水解酶(L-NCC水解酶)激活剂Mn2+和L-半胱氨酸脱巯基酶抑制剂盐酸羟胺来提高L-半胱氧酸产量.结果:以增殖活化10 h的固定化细胞为酶源,DL-ATC·3H2O质量浓度为20g/L,pH 8.0,42℃酶促反应10 h,产物L-半胱氨酸含量达9.18g/L,较游离细胞提高了29.0%,底物转化率达75.83%.固定化细胞重复使用4批次后相对转化率仍能维持在初始值的71.5%.结论:海藻酸钠/明胶协同包埋假单胞菌B-3细胞具有良好的生物转化活性;转化液中添加适量的Tween-60、Mn2+和盐酸羟胺能明显提高L-半胱氨酸产量.     

Pseudomonas sp.ZD-8对菜油降解特性的研究

从土壤中分离一株Psendomonassp .ZD - 8菌株 ,具有较强的降解菜油的能力。在温度为 30℃、pH值为 7、摇床转速 2 10r/min和接种量为 10 %条件下 ,该菌在 2 4h内降解 5 0 0mg/L菜油的去除率达到 95 .6 %;假单胞菌的菜油降解反应符合Monod动力学模式 ,其反应的米氏常数Km为 15 0 1mg/L ,最大反应速度为 80 .4mg菜油 / (10 8.h) .     

Degradation of natural estrogen and identification of the metabolites produced by soil isolates of Rhodococcus sp. and Sphingomonas sp.

Five bacterial strains capable of utilizing 17β-estradiol (E2) and estrone (E1) were isolated from soil samples. Using their morphological and physiological features and 16S rDNA sequences, we classified these isolates into two groups: Group A (Rhodococcus sp. strains ED6, ED7, and ED10) and Group B (Sphingomonas sp. strains ED8 and ED9). All isolates used E2 and E1 as the sole carbon sources and showed high E1 and E2 degradation activities. In all strains, more than 50% of 0.8 mg of E1 or E2 was degraded in 4 mL of inorganic medium over 24 h, and 90% was degraded over 120 h. By incubating the resting ED8 cells with E2 and the meta-cleavage inhibitor 3-chlorocatechol, we identified two metabolites, 4-hydroxyestrone (4-OH-E1) and 4-hydroxyestradiol (4-OH-E2), and confirmed their identity using authentic chemicals. The 4-OH-E1 and 4-OH-E2 compounds were assumed to be intermediate metabolites formed before meta-cleavage, as they were not identified in culture without 3-chlorocatechol. Degradation of E2 by strain ED8 can be initiated by hydroxylation of the C-4 position, followed by meta-cleavage of the benzene ring. When strains ED8 degraded E2, we further identified hydroxy-E2, keto-E1 and -E2, and an additional degradation product via mass spectrometry. The presence of these compounds implied degradation through a second pathway initiated through an attack of the saturated ring.     

Constitutive production of aconitate isomerase by Pseudomonas sp. WU-0701 in relation to trans-aconitic acid assimilation

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Optimization of critical medium components using response surface methodology for phenazine-1-carboxylic acid production by Pseudomonas sp. M-18Q

The optimal flask-shaking batch fermentation medium for phenazine-1-carboxylic acid (PCA) production by Pseudomonas sp. M-18Q, a qscR chromosomal inactivated mutant of the strain M18 was studied using statistical experimental design and analysis. The Plackett-Burman design (PBD) was used to evaluate the effects of eight medium components on the production of PCA, which showed that glucose and soytone were the most significant ingredients (P<0.05). The steepest ascent experiment was adopted to determine the optimal region of the medium composition. The optimum composition of the fermentation medium for maximum PCA yield, as determined on the basis of a five-level two-factor central composite design (CCD), was obtained by response surface methodology (RSM). The high correlation between the predicted and observed values indicated the validity of the model. A maximum PCA yield of 1240 mg/l was obtained at 17.81 g/l glucose and 11.47 g/l soytone, and the production was increased by 65.3% compared with that using the original medium, which was at 750 mg/l.     

生物法降解油烟废气的研究

实验考察了气体停留时间，进气负荷和温度等对油烟去除率的影响，初步研究了生物滤塔的抗冲击能力。结果表明，当停留时间大于20s，温度25～35℃，进气负荷为80～400g／(m^3h)，生物滤塔去除率达到90％以上，且具有较强的抗冲击能力。     

Removal of p-xylene with Pseudomonas sp. NBM21 in biofilter

As a p-xylene (p-Xyl)-degrading microorganism, Pseudomonas sp. NBM21 was isolated from an activated sludge of a wastewater treatment plant. NBM21 degraded p-Xyl, m-xylene, benzene and toluene, but not o-xylene, ethylbenzene (Eb) and styrene. NBM21 was inoculated to a biofilter with Biosol as a packing material and p-Xyl removal was operated for 105 d under sterile and nonsterile conditions. The maximum elimination capacities for p-Xyl at higher than 90% removal efficiency were 160 g/m3/h and 150 g/m3/h under nonsterile and sterile conditions, respectively. A high load of Eb adversely affected to the removal of xylene.     

The increased viability of probiotic Lactobacillus salivarius NRRL B-30514 encapsulated in emulsions with multiple lipid-protein-pectin layers

Probiotics have demonstrated various health benefits but have poor stability to sustain food processing and storage conditions, as well as after ingestion. Biopolymer beads are commonly studied to encapsulate probiotic cells to improve their stability, but the millimeter-dimension of these beads may not meet the quality requirement of food products. The aim of this study was to enhance the viability of Lactobacillus salivarius NRRL B-30514 by encapsulation in emulsion droplets with multiple lipid-protein-pectin layers. Spray-dried L. salivarius was suspended in melted anhydrous milk fat that was then emulsified in a neutral aqueous phase with whey protein isolate or sodium caseinate to prepare primary solid/oil/water (S/O/W) emulsions. Subsequently, pectin was electrostatically deposited onto the droplet surface at pH 3.0 to form secondary emulsions. The encapsulation efficiency was up to 90%. After 20-day storage at 4 °C, the viable cell counts of bacteria in secondary emulsions at pH 3.0 and primary emulsions at 7.0 were 3 log higher than the respective free cell controls. After heating at 63 °C for 30 min, free L. salivarius was inactivated to be undetectable, while about 2.0 log CFU/mL was observed for primary (at pH 7.0) and secondary (at pH 3.0) emulsion treatments. Additionally, a 5 log-CFU/g-powder reduction was observed after spray drying free L. salivarius, while a 2 log CFU/g reduction was observed for emulsion treatments with capsules smaller than 20 μm. Furthermore, cross-linking the secondary emulsion with calcium enhanced the viability of L. salivarius after the simulated gastric and intestinal digestions. Therefore, the studied S/O/W emulsion systems may be used to improve the viability of probiotics during processing, storage, and gastrointestinal digestion.     

Determination of the extracellular and cell-associated hydrolase profiles of Pseudomonas fluorescens sp. using the Analytab API ZYM system

The extracellular and cell-associated hydrolase profiles of a number of Pseudomonas fluorescens strains were examined with the Analytab API ZYM system. Esterase/lipase was the only strong extracellular enzyme activity detected (mean 3.33): weak esterase, lipase, and leucine aminopeptidase activities were found with some strains (mean activities of 1.08, 1.53, and 1.40, respectively). Very strong leucine aminopeptidase activity (4.5) was associated with the cells. Cell-associated trypsin, esterase/lipase, acid phosphatase, and phosphoamidase were also found. Neither extracellular nor cell-associated hydrolase profiles changed significantly when cells were grown in skim milk or mineral salts medium at either 5 or 20 degrees C. Similarly, added calcium did not seem required for synthesis of any of the enzymes. The extracellular enzyme profiles differed considerably from those of the cell-associated enzymes for all strains tested. An extracellular proteinase-deficient mutant of strain 32A (RM14) failed to produce significant quantities of extracellular esterase/lipase activity. Production of cell-associated enzymes was unaffected by the mutation. These results suggest that the Analytab API ZYM system may be useful in identifying psychrotrophs isolated from milk.     

Purification and characterization of an l-amino acid oxidase from Pseudomonas sp. AIU 813

An l-amino acid oxidase was found from a newly isolated strain, Pseudomonas sp. AIU 813. This enzyme was remarkably induced by incubation with l-lysine as a nitrogen source, and efficiently purified using an affinity chromatography with l-lysine as ligand. The enzyme oxidized l-lysine, l-ornithine and l-arginine, but not other l-amino acids and d-amino acids. The oxidase activity for l-lysine was detected in a wide pH range, and its optimal was pH 7.0. In contrast, the oxidase activity for l-ornithine and l-arginine was not shown in acidic region from pH 6.5, and optimal pH for both substrates was 9.0. The enzyme was a flavoprotein and composed of two identical subunits with molecular mass of 54.5?kDa. The N-terminal amino acid sequence was similar to that of putative flavin-containing amine oxidase and putative tryptophan 2-monooxygenase, but not to that of l-amino acid oxidases.     

太平洋杆菌对铜绿假单胞菌毒力因子的抑制作用

为了研究太平洋杆菌XC22919(Oceanobacillus sp.XC22919)对野生型铜绿假单胞菌PAO1(Pseudomonas aeruginosa,PAO1)群体感应(Quorum sensing,QS)系统控制的相关毒素的抑制作用以及抑制效果.以PAO1为检测菌株,在不影响其生长的浓度下,通过吸光度法检测XC22919发酵粗提物对野生型铜绿假单胞菌毒素的抑制率.结果显示:太平洋杆菌XC22919在不影响PAO1生长的浓度下,对其绿脓素、弹性蛋白酶、蛋白水解酶、生物体膜的最高抑制率分别为31％,30％,18％和15 ％.表明太平洋杆菌XC22919发酵液粗提物可以抑制PAO1相关毒素的分泌,具有很好的筛选群体感应抑制剂的潜力,该结果有助于推动新型抗菌药物的研究.     

HPLC-MS/MS同时检测大鼠体内三聚氰胺和三聚氰酸含量

目的建立HPLC-MS/MS同时检测大鼠血浆和组织中三聚氰胺和三聚氰酸含量的方法。方法以同位素标记的三聚氰胺和三聚氰酸作为内标物,大鼠血浆和组织经二氯甲烷-乙腈前处理后,经色谱柱分离,串联离子阱质谱采用ESI源按运行时间段切换正、负离子模式进行测定。结果在0.3～75μg/ml范围内,三聚氰酸和三聚氰胺具有良好的线性(r≥0.998)。在0.9、5.0、50μg/ml的给药大鼠的血浆样本中,三聚氰酸和三聚氰胺的回收率分别为97.4%～99.4%、96.8%～98.9%,两者的变异系数为2.43%～6.35%(日内)、1.70%～4.12%(日间)和0.30%～9.17%(日内)、0.75%～3.39%(日间);在0.9、5.0、50μg/ml的给药大鼠肝组织样本中,三聚氰酸和三聚氰胺的回收率分别为90.8%～104.6%、91.7%～106.0%,对应的变异系数为3.70%～5.14%(日内)、2.84%～4.50%(日间)和1.31%～2.21%(日内)、1.12%～3.36%(日间)。结论该方法样品前处理简单,基质干扰小,分析效率高,具有良好的精密度、准确度和特异性,是进行三聚氰酸、三聚氰胺在动物体内的组织代谢和毒性研究的理想分析方法。     

A preliminary study of caffeine degradation by Pseudomonas sp. GSC 1182

Several microorganisms isolated from soil were tested for their ability to utilize caffeine as the sole carbon and nitrogen source. The isolate identified as Pseudomonas sp. GSC 1182 showed 80% degradation of caffeine in 48 h when caffeine was used as the sole carbon and nitrogen source. In the presence of sucrose (5 g/l), 100% degradation of caffeine was achieved within 36-40 h. The degradation rate was also found to increase when fructose, lactose and galactose were used as carbon source. The isolate showed decreased level (<10%) of caffeine degradation in the presence of glucose. At an initial pH of 6.0, the complete degradation of caffeine was attained in 24 h. The addition of urea and ammonium sulfate as external nitrogen source decreased the caffeine degradation to 35% and 70% respectively.     

Pseudomonadaceae sp.胆固醇氧化酶产酶条件优化及其初步纯化

Pseudomonadaceae sp.可以发酵生产胆固醇氧化酶。本文对Pseudomonadaceae sp.进行产酶条件的优化及产物的初步分离。确定其最适培养基及培养条件,优化后胆固醇氧化酶的酶活力可达1691.72U/L,比优化前839.35U/L有较大提高。经过(NH4)2SO4沉淀及Sephadex G-75凝胶过滤层析初步纯化后酶比活力达24.95U/mg,纯化倍数为9.18。     

Modulator-mediated synthesis of active lipase of Pseudomonas sp. 109 by Escherichia coli cell-free coupled transcription/translation system

Catalytically active lipase was synthesized using Escherichia coli S30 extract from the signal-deleted lipL gene (lipL) in the presence of its N-terminal hydrophobic fragment-truncated modulator (rLimL) that was purified from the overexpressing E. coli cells. The specific activity of the lipase thus synthesized was 125 times higher than that of the purified one from Pseudomonas sp. 109. No lipase activity was detected in the absence of rLimL, even though the lipase protein itself was synthesized. Active lipase was also produced in vitro by coexpression of rlipL and the modulator gene (rlimL), although a much smaller amount of the lipase was formed. In the absence of rLimL, aggregates of the lipase were formed during its folding process. The addition of rLimL proportionally raised both lipase solubility and enzyme activity. An unstable but high activity peak of the lipase was found during its folding process.     

Performance of a styrene-degrading biofilter inoculated with Pseudomonas sp. SR-5

Styrene removal was studied for 3 months in a laboratory-scale biofilter packed with a mixed packing material of peat and ceramic at a ratio of 1 to 1 on a dry-weight basis and inoculated with Pseudomonas sp. SR-5. More than 90% removal efficiency (RE) was attained at 1-140 g/m3/h styrene loads under nitrogen-source limitation. When RE decreased to 70% after 30 d with an increase in styrene load, readdition of SR-5 and washing of the filter packing material restored the RE to more than 90% by maintaining the population of SR-5 at 1-10% of the total cell number. The maximum elimination capacity (EC) by kinetic analysis was estimated to be 290 g/m3/h. High conversion of the removed styrene carbon to CO2, and significantly small production of cell mass from the removed carbon were confirmed.     

响应面法优化假单胞菌产胞外多糖发酵培养基

采用响应面法对假单胞菌（Pseudomonas sp.）FJY5-13生产胞外多糖的发酵培养基进行优化。通过Plackett-Burman试验、最陡爬坡试验及Box-Behnken试验构建回归方程,结果表明,最佳发酵培养基成分为甘油83.0 g/L、酵母浸粉5.7 g/L、NaCl 8.2 g/L、柠檬酸钠5 g/L、（NH4）2SO4 1 g/L、玉米浆粉7.5 g/L,在此条件下,胞外多糖的产量为10.50 g/L,约为优化前多糖产量7.9 g/L的1.3倍。