柑橘皮中果胶的提取工艺条件研究

目的:研究柑橘皮中果胶的提取工艺条件,提高果胶资源的开发及利用度,为农产品深加工提供理论基础。方法:以柑橘皮为原料,采用盐酸提取、真空浓缩、乙醇沉淀的方法,以果胶提取率和吸光度为考察指标,在单因素试验基础上,进行多因素的L16(45)正交试验设计,研究了浸提时间、浸提温度、溶液pH值、乙醇用量和料液比等重要影响因素,对柑橘皮果胶提取率的影响,获得果胶提取最优化组合,建立最佳的果胶提取工艺参数。结果:最佳的工艺条件为提取温度80℃,料液比1:15,提取液pH1.5,浸提时间2h,乙醇用量80%。在此条件下,果胶提取率可达到11.82%。结论:本实验所得的果胶制备最佳工艺参数组合,能获得较理想的橘皮中果胶的提取率。果胶产品各项检测指标均达到或超过国家标准。     

柚果皮中果胶提取的工艺研究

研究了酸种类、提取液pH值、温度、时间、提取液固比等因素对柚皮中果胶提取的影响。通过单因素及L16(45)正交实验,获得了柚子中提取果胶的最佳条件,即pH值1.5～2.0,提取温度100℃,提取时间100min,提取液固比为20。在此条件下果胶得率达14.9%。提取液经浓缩后采用95%乙醇沉淀,所得果胶胶凝度为152°,半乳糖醛酸含量为76.3%,而灰分仅为2.75%。     

Optimization of polysaccharides (ABP) extraction from the fruiting bodies of Agaricus blazei Murill using response surface methodology (RSM)

Response surface methodoloty (RSM) was used to optimize the extraction conditions of polysaccharides (ABP) from the fruiting body of Agaricus blazei. A central composite design (CCD) was used for experimental design and analysis of the results to obtain the optimal processing parameters. Four independent variables such as extraction temperature (°C), ratio of water to raw material, number of extraction, and extraction time (h) were investigated. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and also analyzed by appropriate statistical methods. The 3-D response surface plot and the contour plot derived from the mathematical models were applied to determine the optimal conditions. The optimum extraction conditions were as follows: extraction temperature 91 °C, ratio of water to raw material 14, number of extraction 6, and extraction time 2.1 h. Under these conditions, the experimental value was 65.8 ± 1.42, which is well in close agreement with value predicted by the model.     

Characterization of bioflocculants from biologically aerated filter backwashed sludge and its application in dying wastewater treatment

In this study, the feasibility of bioflocculant extraction from backwashing sludge to reduce its production costs was investigated. Results showed that ultrasound and base treatment could significantly enhance bioflocculant extraction efficiency, however, flocculating activity was affected. It was observed that bioflocculants extracted from sludge of pH 11.0 had no flocculating activity. In contrast, bioflocculants extracted from sludge of pH 5.0, named as M-1, had good flocculating activity. To further study the flocculating activity of M-1, factors such as bioflocculant dosage, temperature and pH of the reaction solution were tested. The optimal conditions were 6.0 mg/l bioflocculant dosage and pH 5.0, at a temperature of 10 °C. Under these conditions, the flocculating rate of kaolin clay was 92.67%. The effectiveness of such bioflocculants in the decolorization of synthetically dyed wastewater was then examined. In flocculating methylene blue and fast blue in aqueous solutions, decolorization efficiency levels were 82.9% and 77.8%, respectively.     

Optimization of aqueous-assisted extraction of polysaccharides from pumpkin (Cucurbita moschata Duch) and their biological activities

The pumpkin pulp contains a greater composition of edible polysaccharides and has reported with excellent biological applications. This research pertains to optimize the extraction of polysaccharides from the fleshy portion of the pumpkin using aqueous assisted extraction (AAE). The result showed that the optimal extraction condition of pumpkin polysaccharide was as follows: extraction temperature at 55 °C, pH 4.5, and enzyme concentration of 4000 µ/g for 80 min. Under the optimal extraction condition, the yield of pumpkin polysaccharide via AAE (15.4) was significantly higher. The biological activities of extracted polysaccharide including α-amylase inhibition (57.41% at 1000 µg/mL) and anti-inflammatory (50.41% at 25 µg/mL) activity increased significantly. Additionally, the antioxidant activities of extracted pumpkin polysaccharides including IC₅₀ values of DPPH and ABTS were 59.87% and 58.74%, respectively. The pumpkin polysaccharide has maximum inhibitory effects against bacterial strains especially for Escherichia coli than that of fungal strains. It is suggested that the aqueous assisted extraction of is a cost-effective promising method to decrease the processing time as well as enhancing extracted polysaccharide yield – times.     

发酵液中多拉菌素的提取和萃取条件研究

采用单因素实验,分别研究提取试剂、发酵液放置时间、pH值和温度对发酵液中多拉菌素提取效果的影响;然后以乙酸乙酯为萃取试剂,研究萃取次数及萃取体积对多拉菌素萃取效果的影响。结果显示,甲醇为最佳提取试剂;发酵液在pH为3～11、温度为20～80℃的条件下放置144 h,多拉菌素均能稳定存在,提取得到的多拉菌素的质量浓度没有显著变化;浓缩提取液液经2倍体积乙酸乙酯萃取2次即可。该条件下多拉菌素的质量浓度和萃取率分别为151.78μg/mL和98.00%。     

响应面优化纤维素酶辅助提取木豆叶总黄酮工艺

为探讨纤维素酶辅助提取木豆叶总黄酮最佳工艺条件,在单因素试验基础上,以酶用量、酶解温度、酶解时间和酶解pH为影响因子,总黄酮得率为响应值,采用Box-behnken中心组合设计建立4个影响因子与总黄酮得率关系的数学模型,进行响应面法分析。结果表明,纤维素酶辅助提取木豆叶总黄酮的最佳工艺条件为：酶用量为8.65 mg,酶解温度为33.88℃,酶解时间为2.02 h,酶解pH为5.02。在最优条件下总黄酮理论得率为4.86%,实测值为4.88%,拟合得到的模型与实际吻合良好。本研究建立的提取工艺条件稳定可靠,为以后木豆叶总黄酮的应用开发提供实验依据。     

豆腐柴叶提取低酯果胶的研究

豆腐柴叶含有丰富的果胶。文中结合盐酸浸提法的基础上,采用醇氨降酯法提取豆腐柴叶的果胶并检测,提取率为部颁19.5%,pH值为2.9,水分和灰分分别为5.13%和8.67%,半乳糖含量为65%,酯化度为38.5%,所提取的低酯果胶的基本性质与国标检测标准接近。果胶成品经AB-8树脂柱纯化后,所提果胶的成分经硅胶板薄层层析初步测定,果胶多糖成分为葡萄糖,果糖,D-甘露糖3种,果胶的溶解度与温度呈正相关性,pH值对其溶解度影响不大。     

Effective technological pectinases by Aspergillus carneus NRC1 utilizing the Egyptian orange juice industry scraps

The production of a notable and highly effective pectinase by the local fungal strain Aspergillus carneus NRC1 utilizing the abundant Egyptian orange peels and pulps (OPP) scraps excluded in the orange juice and canning industry was achieved in 5-day submerged fermentation (SMF) cultures, at temperature and pH ranges of 30–55 °C and 5.0–5.5, respectively. Fresh or thawed OPP (6%, w/v) were the most preferable sole carbon source. Pectinase activity was dramatically stimulated by ammonium sulphate as the sole nitrogen source, and at the same time strongly inhibited the production of the other tested enzymes, i.e., cellulases and hemicellulases. The lyophilized enzyme preparation was free from any ochra or aflatoxins. The optimum conditions of this methodology including enzyme and substrate (citrus pectin) concentrations were 40 mg ml^?1 and 7% (w/v), respectively, with pH and temperature of 4.0 and 55 °C, respectively.     

Enzyme assisted extraction of polysaccharides from the fruit of Cornus officinalis

Process of enzyme assisted extraction (EAE) of polysaccharides from Cornus officinalis was optimized by response surface methodology (RSM). The influence of four different factors on the yield of C. officinalis polysaccharides (COP) was studied. Results showed that the optimal conditions were compound enzyme amount of 2.15%, extraction pH of 4.2, extraction temperature of 55 °C and extraction time of 97 min. Under these conditions, the COP yield was 9.29 ± 0.31%, which was well in agreement with the value predicted by the model. The three methods, EAE, hot water extraction (HWE), ultrasound-assisted extraction (UAE) for extracting COP by RSM were further compared. Results showed that EAE had the largest yield of polysaccharides with lower equipment cost.     

Ultrasound Assisted Extraction of Phenolic Compounds from Peaches and Pumpkins

The ultrasound-assisted extraction (UAE) method was used to optimize the extraction of phenolic compounds from pumpkins and peaches. The response surface methodology (RSM) was used to study the effects of three independent variables each with three treatments. They included extraction temperatures (30, 40 and 50°C), ultrasonic power levels (30, 50 and 70%) and extraction times (10, 20 and 30 min). The optimal conditions for extractions of total phenolics from pumpkins were inferred to be a temperature of 41.45°C, a power of 44.60% and a time of 25.67 min. However, an extraction temperature of 40.99°C, power of 56.01% and time of 25.71 min was optimal for recovery of free radical scavenging activity (measured by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) reduction). The optimal conditions for peach extracts were an extraction temperature of 41.53°C, power of 43.99% and time of 27.86 min for total phenolics. However, an extraction temperature of 41.60°C, power of 44.88% and time of 27.49 min was optimal for free radical scavenging activity (judged by from DPPH reduction). Further, the UAE processes were significantly better than solvent extractions without ultrasound. By electron microscopy it was concluded that ultrasonic processing caused damage in cells for all treated samples (pumpkin, peach). However, the FTIR spectra did not show any significant changes in chemical structures caused by either ultrasonic processing or solvent extraction.     

桔皮果胶提取条件的选择

用酸浸提、酒精沉淀法,从桔皮中提取果胶,通过正交设计分析,获得最佳得率的工艺条件。即果胶浸提液的酸度为pH2,反应时间2小时。沉淀果胶的酒精浓度为50％,其得率是12.6％。     

Maximal release of highly bifidogenic soluble dietary fibers from industrial potato pulp by minimal enzymatic treatment

Potato pulp is a poorly utilized, high-volume co-processing product resulting from industrial potato starch manufacturing. Potato pulp mainly consists of the tuber plant cell wall material and is particularly rich in pectin, notably galactan branched rhamnogalacturonan I type pectin which has previously been shown to exhibit promising properties as dietary fiber. The objective of this study was to solubilize dietary fibers from potato pulp by a one-step minimal treatment procedure and evaluate the prebiotic potential of the fibers. Statistically designed experiments were conducted to investigate the influence of enzyme type, dosage, substrate level, incubation time, and temperature on the enzyme catalyzed solubilization to define the optimal minimal enzyme treatment for maximal fiber solubilization. The result was a method that within 1 min released 75% [weight/weight (w/w)] dry matter from 1% (w/w) potato pulp treated with 1.0% (w/w) [enzyme/substrate (E/S)] pectin lyase from Aspergillus nidulans and 1.0% (w/w) E/S polygalacturonase from Aspergillus aculeatus at pH 6.0 and 60 °C. Molecular size fractionation of the solubilized fibers revealed two major fractions: one fraction rich in galacturonic acid of 10–100 kDa indicating mainly homogalacturonan, and a fraction >100 kDa rich in galactose, presumably mainly made up of β-1,4-galactan chains of rhamnogalacturonan I. When fermented in vitro by microbial communities derived from fecal samples from three healthy human volunteers, both of the solubilized fiber fractions were more bifidogenic than fructo-oligosaccharides (FOS). Notably the fibers having molecular masses of >100 kDa selectively increased the densities of Bifidobacterium spp. and Lactobacillus spp. 2–3 times more than FOS.     

Effects of Temperature, pH, and NaCl on Growth and Pectinolytic Activity of Pseudomonas marginalis

The interaction of temperature (4, 10, 18, and 30°C), pH (6, 7, and 8), and NaCl (0, 2.5, and 5%) and their effects on specific growth rate, lag phase, and pectinolytic enzymes of Pseudomonas marginalis were evaluated. Response surface methodology was adapted to describe the response of growth parameters to environmental changes. To obtain good conditions of storage, the combined action of salt and temperature is necessary. At 4°C with an NaCl concentration of 5% and a pH of 7, the lag time was 8 days and no growth was observed at 4°C with 5% NaCl and a pH of 6. In the absence of salt, P. marginalis could grow regardless of temperature and pH. Pectate lyase and pectin lyase were produced by P. marginalis, while pectin methyl esterase activity was not observed in our culture conditions. The enzyme production depended on temperature, pH, and salt concentration but also on the age of the culture. Pectinolytic enzymes were abundantly excreted during the stationary phase, and even at 4°C, after 2 weeks of storage, enzyme activities in supernatant culture were sufficient to damage vegetables. Both bacterial growth and enzymatic production have to be taken into account in order to estimate correctly the shelf life of vegetables.     

Optimization of pancreatic lipase inhibition by Cudrania tricuspidata fruits using response surface methodology

The fruits of Cudrania tricuspidata (Carr.) Bur. (Moraceae) significantly inhibited pancreatic lipase, which plays a key role in fat absorption. Optimization of extraction conditions with minimum pancreatic lipase activity and maximum yield was determined using response surface methodology with three-level-three-factor Box–Behnken design (BBD). Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as ethanol concentration, 74.5%; temperature 61.9 °C and extraction time, 13.5 h. The pancreatic lipase activity and extraction yield under optimal conditions were found to be 65.5% and 54.0%, respectively, which were well matched with the predicted value of 65.8% and 47.1%. Further fractionation of C. tricuspidata extract resulted in the isolation of compound 1, which was identified as 5,7,4′-trihydroxy-6,8-diprenylisoflavone. It inhibited pancreatic lipase activity with IC₅₀ value of 65.0 μM. HPLC analysis suggested positive correlation between pancreatic lipase inhibition and 5,7,4′-trihydroxy-6,8-diprenylisoflavone of C. tricuspidata fruits.     

Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization

Lignocellulose is the major component of plant cell walls and it represents a great source of renewable organic matter. One of lignocellulose constituents is pectin. Pectin is composed of two basic structures: a ‘smooth’ region and a ‘hairy’ region. The ‘smooth’ region (homogalacturonan) is a linear polymer of galacturonic acid residues with α-(1→4) linkages, substituted by methyl and acetyl residues. The ‘hairy’ region is more complex, containing xylogalacturonan and rhamnogalacturonans I and II. Among the enzymes which degrade pectin (pectinases) is pectin lyase (E.C. 4.2.2.10). This enzyme acts on highly esterified homogalacturonan, catalysing the cleavage of α-(1→4) glycosidic bonds between methoxylated residues of galacturonic acid by means of β-elimination, with the formation of 4,5-unsaturated products. In this work, the gene and cDNA of a pectin lyase from Penicillium purpurogenum have been sequenced, and the cDNA has been expressed in Pichia pastoris. The gene is 1334 pb long, has three introns and codes for a protein of 376 amino acid residues. The recombinant enzyme was purified to homogeneity and characterized. Pectin lyase has a molecular mass of 45 kDa as determined by SDS-PAGE. It is active on highly esterified pectin, and decreases 40 % the viscosity of pectin with a degree of esterification ≥85 %. The enzyme showed no activity on polygalacturonic acid and pectin from citrus fruit 8 % esterified. The optimum pH and temperature for the recombinant enzyme are 6.0 and 50 °C, respectively, and it is stable up to 50 °C when exposed for 3 h. A purified pectin lyase may be useful in biotechnological applications such as the food industry where the liberation of toxic methanol in pectin degradation should be avoided.     

Quantification of 4-aminopyridine in plasma by capillary electrophoresis with electrokinetic injection

A rapid and sensitive CE method for the determination of 4-aminopyridine in human plasma using 3,4-diaminopyridine as an internal standard was developed and validated. The analytes were extracted from 0.5-mL aliquots of human plasma by liquid–liquid extraction, using 8 mL of ethyl ether, and injected electrokinetically into capillary electrophoresis equipment. The instrumental conditions were obtained and optimized by Design of Experiments (DOE – factorial and response surface model), having as factors: separation voltage, ionic strength (buffer concentration), pH and temperature. The response variables were migration time, resolution, tailing factor and drug peak area. After obtaining mathematically predicted values for the response variables with best factors combinations, these were reproduced experimentally in good agreement with predicted values. In addition to optimal separation conditions obtained by Design of Experiments, sensitivity was improved using electrokinetic injection at 10 kV for 10 s, and a capillary with 50 cm effective length and 100 μm I.D. The final instrumental conditions were voltage at 19 kV, capillary temperature at 15 °C, wavelength at 254 nm, and phosphate buffer 100 mM, pH 2.5 as the background electrolyte. This assay was linear over a concentration range of 2.5–80 ng/mL with a lower limit of quantification of 2.5 ng/mL. The relative standard deviation for the assay precision was <7% and the accuracy was >95%. This method was successfully applied to the quantification of 4-aminopyridine (4-AP) in plasma samples from patients with spinal cord injury.     

Effects of temperature on simultaneous nitrification and denitrification via nitrite in a sequencing batch biofilm reactor

A laboratory scale experiment was described in this paper to enhance biological nitrogen removal by simultaneous nitrification and denitrification (SND) via nitrite with a sequencing batch biofilm reactor (SBBR). Under conditions of total nitrogen (TN) about 30 mg/L and pH ranged 7.15–7.62, synthetic wastewater was cyclically operated within the reactor for 110 days. Optimal operation conditions were established to obtain consistently high TN removal rate and nitrite accumulation ratio, which included an optimal temperature of 31 °C and an aeration time of 5 h under the air flow of 50 L/h. Stable nitrite accumulation could be realized under different temperatures and the nitrite accumulation ratio increased with an increase of temperature from 15 to 35 °C. The highest TN removal rate (91.9%) was at 31 °C with DO ranged 3–4 mg/L. Process control could be achieved by observing changes in DO and pH to judge the end-point of oxidation of ammonia and SND.     

Quantification of phototrophic biomass on rocks: optimization of chlorophyll-a extraction by response surface methodology

Biological colonization of rock surfaces constitutes an important problem for maintenance of buildings and monuments. In this work, we aim to establish an efficient extraction protocol for chlorophyll-a specific for rock materials, as this is one of the most commonly used biomarkers for quantifying phototrophic biomass. For this purpose, rock samples were cut into blocks, and three different mechanical treatments were tested, prior to extraction in dimethyl sulfoxide (DMSO). To evaluate the influence of the experimental factors (1) extractant-to-sample ratio, (2) temperature, and (3) time of incubation, on chlorophyll-a recovery (response variable), incomplete factorial designs of experiments were followed. Temperature of incubation was the most relevant variable for chlorophyll-a extraction. The experimental data obtained were analyzed following a response surface methodology, which allowed the development of empirical models describing the interrelationship between the considered response and experimental variables. The optimal extraction conditions for chlorophyll-a were estimated, and the expected yields were calculated. Based on these results, we propose a method involving application of ultrasound directly to intact sample, followed by incubation in 0.43 ml DMSO/cm² sample at 63°C for 40 min. Confirmation experiments were performed at the predicted optimal conditions, allowing chlorophyll-a recovery of 84.4 ± 11.6% (90% was expected), which implies a substantial improvement with respect to the expected recovery using previous methods (68%). This method will enable detection of small amounts of photosynthetic microorganisms and quantification of the extent of biocolonization of stone surfaces.     

娄彻氏链霉菌ATCC10739产抗生素Borrelidin发酵条件优化及其分离纯化

对娄彻氏链霉菌ATCC10739固体发酵产生十八元大环内酯抗生素Borrelidin进行了发酵条件的优化。首先筛选得到了理想的发酵培养基;其次考察了发酵时间、起始pH值以及在ISP-2培养基中添加附加碳源、氮源对Borrelidin产量的影响。初步确定最适发酵条件为:ISP-2培养基中添加1%甘油,起始pH值为6.0,培养温度30°C,发酵时间为7 d,产量可达1.336 mg/L。采用有机溶剂萃取、硅胶层析和半制备型高效液相色谱(HPLC)等分离技术纯化得到Borrelidin。