环境因子对鳗弧菌生物膜形成的影响

【背景】鳗弧菌是海产动物弧菌病的主要病原,在海水水域中广泛存在。鳗弧菌为了适应环境变化会生成生物膜,形成自我保护,对其防治是水产养殖行业的一大难题。【目的】探讨致病性鳗弧菌(Vibrio anguillarum)BYK0638生物膜的形成特性,为进一步研究鳗弧菌生物膜形成机制和致病机理提供参考。【方法】采用改良的微孔板法研究静置培养条件下鳗弧菌(V.anguillarum)BYK0638在96孔酶标板上的成膜情况,CCK-8法(Cell counting kit-8)定量检测生物膜中鳗弧菌的活力。【结果】鳗弧菌BYK0638能够在聚苯乙烯酶标板上形成稳定而明显的生物膜,其生物膜的OD450值在24 h达到峰值,60 h后趋于稳定;在107-108 CFU/m L范围内,鳗弧菌生物膜的OD450值显著高于其他试验组(P0.05);25°C时的生物膜OD450值显著高于其他温度生物膜的形成量;在p H 4.0-11.0范围内,当p H值为7.0时鳗弧菌形成的生物膜量最高,在p H值为3.0和12.0时鳗弧菌几乎不形成生物膜;在TSB培养基中加入0.03-2.00 mmol/L Ca Cl2,鳗弧菌生物膜形成量与未添加Ca Cl2对照组无显著性差异;在TSB培养基中加入0.03 mmol/L Mg Cl2,可促进鳗弧菌生物膜形成;Na Cl浓度为5%时,形成的生物膜OD450值最高;鳗弧菌在大黄鱼表皮黏液、肝脏、前肠、后肠组织提取液包被的96孔酶标板上形成的生物膜显著高于其他黏液和组织提取液包被组(P0.05)。【结论】致病性鳗弧菌BYK0638能形成稳定而明显的生物膜,其生物膜形成与外界环境因子变化有密切的关系,培养时间、初始菌浓度、温度、p H、Mg2+、盐度及不同组织和黏液等各种环境因子均能显著影响鳗弧菌生物膜的形成。     

酶法提取薏苡仁油的优化

本文应用响应面分析的方法研究了酶用量、酶解温度和pH对薏苡仁油提取率的影响,并得到了最佳的提取条件。研究结果表明,酶用量为1.569%、酶解温度在47.7℃、酶解pH为4.75。响应的薏苡仁油提取率为10.948%。油脂质量分析结果表明,酶法处理并没有显著改变薏苡仁油的性质。     

疫苗

知12,‘用二步酶水解法由整鱼制备蛋白膝及这种蛋白膝的特性瑛}/Gildberg,A.…/l BioteChnol.APPI.Biochern一l,8夕,11(4)一4 13~423降自DBA,1989,8(19),89一1 1298】 两步水解法包括鱼料自溶3天,然后在碱性条件下,用中性鱼蛋白酶困FP)或尿黑酶(Alcalase)进行水解.两种酶都能有效地消化毛鳞鱼,但尿黑酶消化兰牙彗鱼更有效.虽然在兰牙彗鱼水解时,尿黑酶比中性蛋白酶产生更高的N,但中性蛋白陈的终产量相同.最终的蛋白陈溶液的超过滤实验证明,大约70%粗蛋白以游离氮基酸或分子量1 000以下的肤存在.这些蛋白脉用于细菌生长实验(对鳗弧菌,…     

超声辅助水酶法提取月见草籽油的研究

月见草籽油在医药、保健及食品工业领域具有重要价值。本文对超声辅助水酶法提取月见草籽油进行了研究,采用超声及粉碎处理方式对月见草籽进行预处理,确定最佳预处理条件：超声功率300W、超声时间30min、超声温度60℃;采用Alcalase 2.4L碱性蛋白酶进行酶解,利用响应面优化试验,确定最优的月见草籽油脂提取工艺：料液比为5.4（w/w）、酶添加量为1.38（v/w）、酶解温度为62.5℃、酶解时间2.8h,响应面有最优值为84.32%;测定油脂的基本组成及相关品质指标,结果表明,不同提取方法对月见草籽油的皂化值、折射率、色泽的影响不显著,但酸值及磷脂含量均低于溶剂法月见草籽油。优化的工艺简便可行、提取率高,为超声辅助水酶法提取月见草籽油提取工艺的产业化提供理论依据。     

不同方式提取松籽油理化性质及微波加热对其品质影响的研究

油脂氧化是导致食品质量下降的主要原因。油脂在微波加热过程品质会发生变化,本研究旨在确定微波加热过程对不同方式提取的松籽油的影响。本文对水酶法和溶剂法提取的松籽油的理化性质及脂肪酸组成进行了分析,两种方法提取的松籽油品质有一定的不同,水酶法松籽油酸值及磷脂含量均低于溶剂法松籽油,而过氧化值高于溶剂法松籽油,提取方法对松籽油脂肪酸组成的影响不显著。同时,研究了在一定功率（700W）下,不同加热时间（1、3、5和7min）对两种方法提取松籽油品质的影响,微波加热3min以上,会导致所有样品品质恶化及组成变化。     

水酶法提取丝瓜籽油的工艺研究及其理化性质分析

通过单因素和正交试验,确定水酶法提取丝瓜籽油的最佳工艺,并对得到的油脂的理化性质进行测定。结果表明,最佳工艺为酶解时间6.5h、酶解温度50℃、酶添加量5%、p H6,在此条件下丝瓜籽提油率可达29.6%。水酶法提取的丝瓜籽油各项理化性质均符合国家标准,且比传统工艺提取的丝瓜籽油品质更好。     

大豆种子蛋白和油脂含量调控的研究进展

作为重要的粮油饲兼用作物,大豆为世界膳食提供高达约71%的蛋白质和29%的油脂。随着人口不断增长和大豆消费需求的不断提高,在有限的耕地面积和单产条件下,大豆品质的遗传改良则更具重要意义。该文综述了大豆种子蛋白和油脂含量两个重要品质性状调控的研究进展,总结了调控大豆蛋白和油脂合成的关键酶和转录因子及因子间的相互作用,并根据蛋白和油脂合成代谢调控途径中关键酶和转录因子作用机制,绘制了大豆蛋白和油脂合成代谢的分子调控网络。此外,该文还讨论了当前大豆种子蛋白油脂含量调控研究存在的瓶颈及对策,以期为大豆种子品质的遗传改良和高产品种培育提供参考。     

恒化培养稀释率和碳氮比对圆红冬孢酵母油脂积累的影响

采用恒化培养的方法,考察了稀释率(D)和碳氮比(mol/mol)对圆红冬孢酵母Rhodosporidiumtoruloides AS 2.138 9积累油脂的影响。结果表明:稀释率增大,油脂含量和油脂得率降低。在D=0.02 h 1时油脂得率最大,为0.18 g油/g糖;D=0.14 h 1时油脂生成速率最大,为0.09 g/(L.h)。碳氮比增大,油脂含量略有增加。在C/N=92时油脂得率最大,为0.12 g油/g糖;C/N=32时油脂生成速率最大,为0.13 g/(L.h)。碳氮比对油脂的脂肪酸组成影响不明显,油脂的棕榈酸、硬脂酸和油酸总含量超过85%。     

人工养殖中华眼镜蛇(Naja naja)油的成分鉴定与分析

目的分析与鉴定人工养殖中华眼镜蛇(Naja naja)油的成分。方法采用酸水解法处理蛇油,用气相色谱-质谱(GC-MS)联用方法,分析与鉴定蛇油的脂肪酸成分。结果采用酸水解法,当水解温度为80℃、石油醚为萃取溶剂时,从蛇油中获得油脂的收率为90.10%;当水解温度为60℃、石油醚为萃取溶剂时,蛇油中多不饱和脂肪酸的含量为21.15%。结论人工养殖的中华眼镜蛇油含有较多生理活性物质,如花生四烯酸、DHA、DPA等。     

双低油菜品系含油量与发育种子中PEPc酶和PAL酶活性的相关分析

以10个含油量不同的双低油菜品系为材料,测定了发育种子中磷酸烯醇式丙酮酸羧化酶（PEPc）和苯丙氨酸解氨酶（PAL）活性动态变化,以及成熟种子的油脂含量,分析了PEPc和PAL酶活性与含油量之间的关系。结果表明：PEPc酶与含油量之间始终存在不显著正相关,说明PEPc酶恬性高低对含油量高低有一定作用,但并不是油菜种子油脂合成过程中的关键酶;PAL酶与含油量之间始终存在不显著的负相关,授粉后32d,两者之间负相关性最高,但该相关性并不显著。     

Enzymatic hydrolysis of cod (Gadus morhua) by-products: Optimization of yield and properties of lipid and protein fractions

The main products of hydrolysis of fish by-products are hydrolysed protein and oil. The aim of this work was to study the effect of initial heat inactivation of endogenous enzymes, addition of water prior to hydrolysis, use of different commercial enzymes and combination of enzymes on the yield and purity of the protein and oil fractions after enzymatic hydrolysis of cod by-products. This study was designed to examine how all these factors were effective for destroying protein–lipid complexes in order to obtain pure oil and protein fractions and reduce the insoluble fraction.

Initial heating of raw material changed both raw material properties and inactivated endogenous enzymes thereby influencing the following hydrolysis. High amount of lipids in raw material combined with initial heating caused formation of protein–lipid complexes which was found in all protein containing fractions. The main constituents of the lipids in the complexes were phospholipids and other polar lipids. Insoluble protein–lipid complexes formed lead to increased amount of sludge, reduced FPH yield and high amount of lipids in FPH. The highest amount of separated oil was obtained in the experiments after initial heating without added water. These treatments also reduce amount of emulsion, which is not a desirable product after hydrolysis. Initial heating caused denaturation of protein, which decreased their emulsifying properties.

Results showed that it is not possible to obtain all desirable quality indicators such as: maximum oil and FPH yield, minimum emulsion and sludge yield and the highest protein recovery in FPH with the lowest amount of lipids in FPH fraction by using only one hydrolysis process. Therefore, the aim and requirements for the final products should be prioritised and defined very clearly before the process is designed taking into account the composition of raw material. Hydrolysis of unheated raw material with Alcalase and addition of water was the best compromise taking into account the mentioned quality indicators.     

Optimization of lipase-catalyzed biodiesel by response surface methodology

Biodiesel prepared by catalyzed mild transesterification has become of much current interest for bioenergy. The ability of a commercial immobilized lipase (Novo Industries--Bagsvaerd, Denmark) from Rhizomucor miehei (Lipozyme IM-77) to catalyze the transesterification of soybean oil and methanol was investigated in this study. Response surface methodology and 5-level-5-factor central composite rotatable design were employed to evaluate the effects on reaction time, temperature, enzyme amount, molar ratio of methanol to soybean oil, and added water content on percentage weight conversion to soybean oil methyl ester by transesterification. Based on ridge max analysis, the optimum synthesis conditions giving 92.2% weight conversion were: reaction time 6.3 h, temperature 36.5 degrees C, enzyme amount 0.9 BAUN (Batch Acidolysis Units NOVO), substrate molar ratio 3.4:1, and added water 5.8%.     

Soybean oil methyl esters preparation using NaX zeolites loaded with KOH as a heterogeneous catalyst

The transesterification of soybean oil with methanol to methyl esters was carried out using NaX zeolites loaded with KOH as a solid base catalyst. Best result was obtained with NaX zeolite loaded with 10% KOH, followed by heating at 393 K for 3 h. When the transesterification reaction was carried out at reflux of methanol (338 K), with a 10:1 molar ratio of methanol to soybean oil, a reaction time of 8 h and a catalyst amount of 3 wt.%, the conversion of soybean oil was 85.6%.     

Biodiesel production through lipase catalyzed transesterification: An overview

Recently, with the global shortage of fossil fuels, excessive increase in the price of crude oil and increased environmental concerns have resulted in the rapid growth in biodiesel production. The central reaction in the biodiesel production is the transesterification reaction which could be catalyzed either chemically or enzymatically. Enzymatic transesterification has certain advantages over the chemical catalysis of transesterification, as it is less energy intensive, allows easy recovery of glycerol and the transesterification of glycerides with high free fatty acid contents. Limitations of the enzyme catalyzed reactions include high cost of enzyme, low yield, high reaction time and the amount of water and organic solvents in the reaction mixture. Researchers have been trying to overcome these limitations in the enzyme catalyzed transesterification reaction. This paper is meant to review the latest development in the field of lipase catalyzed transesterification of biologically derived oil to produce biodiesel.     

Tissue-specific biomass recalcitrance in corn stover pretreated with liquid hot-water: SEM imaging (part 2)

In the first part of our work, we combined compositional analysis, pretreatment and enzyme hydrolysis for fractionated pith, rind, and leaf tissues from a hybrid stay-green corn, in order to identify the role of structural characteristics on enzyme hydrolysis of cell walls. Hydrolysis experiments coupled with chemical analysis of the different fractions of corn stover showed significant differences in cell wall structure before and after liquid hot water pretreatment. The extent of enzyme hydrolysis followed the sequence rind < leaves < pith with 90% conversion of cellulose to glucose in 24 h in the best cases. Since similar lignin contents remained after liquid hot water pretreatment of leaves, rind, and pith, our results indicated that the amount of lignin alone is not sufficient to explain the different enzymatic hydrolysis characteristics of the fractions. While the role of structural characteristics on enzyme hydrolysis of cell walls is measured as described in part I, the SEM images presented in this part II of our work show that sugar yields from enzymatic hydrolysis of corn fractions correlate with changes in plant cell wall structure both before and after liquid hot water pretreatment.     

Enrichment of polyunsaturated fatty acids from tuna oil using immobilized Pseudomonas fluorescens lipase

Immobilized Pseudomonas fluorescens lipase enzyme was used to enrich the important polyunsaturated fatty acid (PUFA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) from tuna oil. Hydrolysis, esterification, and transesterification reactions were studied in detail to find out the fractionation pattern of DHA and EPA during these processes due to preferential selectivity for or against these PUFA. Hydrolysis with P. fluorescens biotype I lipase with stoichiometric amount of water content gave more than 80% of DHA and EPA in the free fatty acid (FFA) form after around 60% of hydrolysis. After some preferential specificity during the early stages of hydrolysis, P. fluorescens lipase exhibits nonselective characteristics on extended hydrolysis. Esterification of FFA extracted from the completely hydrolyzed mixture of tuna oil was found to be better with long chain fatty alcohol like octanol which lead to good enrichment (44.5% for DHA and 11.3% for EPA) and yields of the PUFA in the FFA form. Transesterification (ethanolysis) with immobilized P. fluorescens lipase enzyme resulted in good enrichment and recovery of DHA and EPA in the glyceride mixture. After around 60% of ester synthesis, 74% of (DHA + EPA) enrichment was achieved with yields of more than 90% in the glyceride mixture.     

Optimization of Pseudomonas cepacia lipase preparations for catalysis in organic solvents

The activity of different lipase (from Pseudomonas cepacia) forms, such as crude powder (crude PC), purified and lyophilized with PEG (PEG + PC), covalently linked to PEG (PEG-PC), cross-linked enzyme crystals (CLEC-PC), and immobilized in Sol-Gel-AK (Sol-Gel-AK-PC) was determined, at various water activities (aw), in carbon tetrachloride, benzene and 1,4-dioxane. The reaction of vinyl butyrate with 1-octanol was employed as a model and both transesterification (formation of 1-octyl butyrate) and hydrolysis (formation of butyric acid from vinyl butyrate) rates were determined. Both rates depended on the lipase form, solvent employed, and aw value. Hydrolysis rates always increased as a function of aw, while the optimum of aw for transesterification depended on the enzyme form and nature of the solvent. At proper aw, some lipase forms such as PEG + PC, PEG-PC, and Sol-Gel-AK-PC had a total activity in organic solvents (transesterification plus hydrolysis) which was close to (39 and 48%) or even higher than (130%) that displayed by the same amount of lipase protein in the hydrolysis of tributyrin-one of the substrates most commonly used as standard for the assay of lipase activity-in aqueous buffer. Instead, CLEC-PC and crude PC were much less active in organic solvents (2 and 12%) than in buffer. The results suggest that enzyme dispersion and/or proper enzyme conformation (favored by interaction with PEG or the hydrophobic Sol-Gel-AK matrix) are essential for the expression of high lipase activity in organic media.     

Biodiesel production from heterotrophic microalgal oil

The present study introduced an integrated method for the production of biodiesel from microalgal oil. Heterotrophic growth of Chlorella protothecoides resulted in the accumulation of high lipid content (55%) in cells. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane. Biodiesel comparable to conventional diesel was obtained from heterotrophic microalgal oil by acidic transesterification. The best process combination was 100% catalyst quantity (based on oil weight) with 56:1 molar ratio of methanol to oil at temperature of 30 degrees C, which reduced product specific gravity from an initial value of 0.912 to a final value of 0.8637 in about 4h of reaction time. The results suggested that the new process, which combined bioengineering and transesterification, was a feasible and effective method for the production of high quality biodiesel from microalgal oil.     

Characteristics of immobilized lipase-catalyzed hydrolysis of olive oil of high concentration in reverse phase system

Candida rugosa lipase immobilized by adsorption on swollen Sephadex LH-20 could almost completely hydrolyze 60% (v/v) olive oil in isooctane. Kinetic analysis of the lipase-catalyzed hydrolysis reaction was found to be possible in this system. Amount of fatty acids produced was linearly proportional to the enzyme concentration of 720 mug/g wet gel. The specific enzyme activity was 217 units/mg protein at 60% (v/v) olive oil concentration. When the initial rate is plotted versus concentration of olive oil, this system did not follow Michaelis-Menten kinetics. Maximum activity was obtained at pH 7, but optimum temperature shifted towards higher one with the increase of olive oil concentration. Among the various chemical compounds tested, Hg(2+) and Fe(2+) inhibited the lipase seriously. As the concentration of olive oil increased, the rate of the hydrolysis also increased, but degree of the hydrolysis was observed to decrease. The supply of water from the inside of the gel to the surface of the gel was the main factor for the control of the rate of hydrolysis in batch hydrolysis. The immobilized lipase was used to hydrolyze olive oil two times. Achievement of chemical equilibrium took a longer time with the addition of water and the degree of hydrolysis decreased in the second consecutive trial. After the second hydrolysis trial, the gels were regenerated in a packed column first by eluting out both residual fatty acids around the gel particles and the accumulated glycerol with ethanol and then with 0.05M phosphate buffer, pH 7. The immobilized lipase on the regenerated gel showed the same hydrolysis activity as the original one.     

Synthesis of antioxidant propyl gallate using tannase from Aspergillus niger van Teighem in nonaqueous media

Tannase from Aspergillus niger van Teighem has been used for synthesis of food additive antioxidant propyl gallate by direct transesterification of tannic acid. The optimized yield of 86% was obtained by using simultaneously pH tuned enzyme, immobilized on Celite and using the right amount of water in the non aqueous media.