PB试验和BBD响应面优化酶解鱼鳞制备抗菌肽工艺

以酶解液的多肽含量和抑菌活性作为考察指标,采用酸性蛋白酶酶解制备草鱼鱼鳞抗菌肽。在单因素的基础上,运用PB试验设计进行影响因素的显著性筛选,确定出显著性影响因素为pH、温度和时间,进而通过BBD试验得到草鱼鱼鳞抗菌肽制备的最佳工艺条件为:pH 2.72、40.22℃、97.33min,在该条件下酶解液对金黄色葡萄球菌的抑菌圈直径为18.49mm,对大肠杆菌的抑菌圈直径为17.31 mm。结果表明,利用酸性蛋白酶酶解制备草鱼鱼鳞抗菌肽是行得通的,为草鱼鱼鳞等下脚料实现低值原料高值化提供了新的思路。     

θ3 fatty acids in freshwater fish from south brazil

Lipid and fatty acid levels in the edible flesh of 17 freshwater fish from Brazil’s southern region were determined. Analyses of fatty acid methyl esters were performed by gas chromatography. Palmitic acid (C_16:0) was the predominant saturated fatty acid, accounting for 50–70% of total saturated acids. Oleic acid (C_18:1θ9) was the most abundant monounsaturated fatty acid. Linoleic acid (C_18:2θ6), linolenic acid (C_18:3θ3), and docosahexaenoic acid (C_22:6θ3) were the predominant polyunsaturated fatty acids (PUFA). The data revealed that species such as truta, barbado, and corvina were good sources of eicosapentaenoic acid (C_20:5θ3) and docosahexaenoic acid (C_22:6θ3), and that most freshwater fish examined were good sources of PUFA θ3.     

A hybrid membrane-emulsion reactor for the enzymatic hydrolysis of lipids

A hybrid reactor, consisting of a stirred vessel, a hydrophilic membrane loop and a hydrophobic membrane loop, is presented for the continuous enzymatic hydrolysis of soybean oil in an emulsion. The permeates of the hydrophilic and the hydrophobic membrane consist of a single water phase and a single lipid phase, respectively. No lipase activity could be detected in the permeates of both membranes, which implies that all enzyme is retained in the system. An important advantage of this system is that it combines the high surface area in an emulsion with the containment of lipase in a membrane reactor. It is further shown that the stability of the system can be improved considerably by the addition of CaCl₂ to the water phase. Under comparable conditions the enzyme stability in the hybrid reactor is lower than the stability in a stirred vessel. The composition of the emulsion appears to influence the flux of the membranes. The flux of the hydrophobic membrane increases with an increasing oil fraction of the emulsion while the flux of the hydrophilic membrane has an optimum for two different oil fractions—0 and 0.55 (v/v).     

Fatty acid composition and oxidation of lipids in Korean catfish

This study determined the lipid content and FA composition of muscle and a mixture of muscle and viscera from Korean catfish as well as lipid oxidation and hydrolysis. Lipid content and FA compositions in Korean catfish, which were purchased every month or two during September 1999–July 2000, were analyzed. Lipid oxidation and hydrolysis were determined as PV, thiobarbituric acid value, and FFA in the muscle and the mixture during storage at 2°C for 12 d and −14°C for 9 wk. Lipid contents of the muscle and the mixture were 3.2 (w/w) and 5.4%, respectively. Oleic acid was the most abundant FA in the catfish lipids, constituting 28.0% (w/w) of total FA in the muscle and 25.6% in the mixture, followed by palmitic acid and linoleic acid, amounting to 20.2 and 12.2%, respectively, in the muscle and 20.2 and 12.5% in the mixture. EPA and DHA were 3.2 and 6.8%, respectively, in the muscle and 3.5 and 8.1% in the mixture. Seasonal variation in lipid contents and FA composition was minimal in catfish. Lipids in minced catfish oxidized and hydrolyzed readily at 2°C. Inclusion of viscera into the muscle increased lipid oxidation and hydrolysis. Frozen storage at −14°C and addition of ascorbic acid both reduced lipid oxidation. Frozen storage retarded lipid hydrolysis in catfish.     

酶/酸水解毛叶木姜子中键合态香味成分的比较

采用AmberliteXAD-2树脂吸附洗脱分离毛叶木姜子中的键合态香气物质前体,将分离得到的键合态香气物质前体在AR2000酶、果胶酶和酸3种条件下分别进行水解释放,采用气相色谱-质谱联用法(GC-MS)对水解后的键合态香气物质进行分离分析。结果表明:3种水解条件得到的键合态香气物质共有90种,主要为萜烯类和含氧萜类物质。其中,香叶醇在2种酶水解条件下均含量高,果胶酶水解条件下含量最高达21798.79μg/L;共34种香气物质具有明显香气特征,2种酶水解下最显著的香气特征为花香和甜香;酸水解条件下最显著的香气特征为花香。酶水解与酸水解条件下得到的键合态香气物质存在明显差异,果胶酶水解条件下所检出的香气物质种类多且含量较高。因此,果胶酶处理更有利于毛叶木姜子键合态香气成分的释放。     

Interference of polar lipids with the alkalimetric determination of free fatty acid in fish lipids

An examination of the suitability of an alkalimetric method for the determination of free fatty acid (FFA) contents in fats, oils, and lipid extracts was conducted by comparing AOCS method Ca 5a-40 with a method based on a Chromarod-latroscan thin-layer chromatography-flame-ionization detector (TLC-FID) system. The FFA contents determined by the alkalimetric method were consistently higher than the genuine FFA contents obtained by the latroscan TLC-FID method. Phospholipids were found to be the major components that contributed to the alkali-titratable, nongenuine FFA in the total FFA determined alkalimetrically. Contributions from other polar lipid components were smaller, but they dominated as the proportion of phospholipids fell. The other alkali-titratable polar components may include oxidized lipids and their by-products bound to protein fragments. The accurate determination of FFA contents by alkalimetric methods may only be applicable to those commercially refined fats and oils that contain negligible amounts of phospholipids. Corrections for the alkalimetrically determined FFA contents should be made for those fats and oils with relatively high phospholipid contents by correlating the nongenuine FFA contents and the phospholipid contents.     

Effect of several factors on peracetic acid pretreatment of sugarcane bagasse for enzymatic hydrolysis

BACKGROUND: Lignocellulose should undergo pretreatment to enhance its enzymatic digestibility before being saccharified. Peracetic acid (PAA) is a strong oxidant that can remove lignin under mild conditions. The sulfuric acid in the PAA solution also can cause degradation of hemicelluloses. The objective of the present work is to investigate the effect of several factors on peracetic acid pretreatment of sugarcane bagasse. RESULTS: It was found that PAA charge, liquid/solid (l/s) ratio, temperature, time, interactions between PAA charge and l/s ratio, temperature and time, all had a very significant effect on the enzymatic conversion ratio of cellulose. The relative optimum condition was obtained as follows: PAA charge 50%, l/s ratio 6:1, temperature 80 °C and time 2 h. More than 80% of the cellulose in bagasse treated under the above conditions was converted to glucose by cellulase of 20 FPU g^?1 cellulose. Compared with H₂SO₄ and NaOH pretreatments under the same mild conditions, PAA pretreatment was the most effective for enhancement of enzymatic digestibility. CONCLUSION: PAA pretreatment could greatly enhance the enzymatic digestibility of sugarcane bagasse by removing hemicelluloses and lignin, but removal of lignin was more helpful. This study can serve as a step to further optimization of PAA pretreatment and understanding the mechanism of enhancement of enzymatic digestibility. Copyright © 2007 Society of Chemical Industry     

酶法提取小檗碱工艺研究

用正交实验法研究了酶法提取小檗碱的最佳工艺条件。以小檗碱提取率为指标,分别考察了温度、时间、酶解pH值、加酶量对酶解反应的影响。确定了酶解黄连的最佳工艺条件:温度40℃、时间90 m in、pH 4.0、酶用量30 mg/g。在此条件下,对酶法提取小檗碱与传统乙醇浸提法进行比较,结果表明,酶法提取工艺比传统乙醇浸提法小檗碱产量提高了49%。     

蔗渣酶法水解过程的酶吸附和活性

以纸浆甘蔗渣纤维素为底物,使用纤维素酶为催化剂,在50℃,pH为4.8的条件下研究了釜式及固定床反应流程,考察酶的吸附及活性.结果表明,固定床反应器的酶吸附比釜式多,而酶的活性变化在两种型式反应器中比较接近.     

甘蔗渣的酸催化常压甘油有机溶剂预处理及其酶解

预处理可以打破木质纤维素原料纤维素、半纤维素和木质素三大组分间的顽抗结构,从而提升纤维素基质可酶解性。本文针对目前常压甘油有机溶剂预处理花费时间过长的问题,尝试开展酸催化的常压甘油有机溶剂预处理研究以缩短预处理时间。实验通过单因素选择和响应面Box-Behnken设计优化,获得酸催化常压甘油有机溶剂预处理的最佳条件为：预处理温度245℃,预处理时间38min,硫酸添加质量0.1%。在此条件下获得基质48h酶解率的响应面预测值为94.0%,实际值为91.4%。结果表明响应面优化方案和回归模型适用于本实验,预处理显著提高了基质可酶解性。高浓度基质（15%～20%）酶解进一步证明了预处理后基质具有突出的可酶解性,20%浓度基质在酶载量5FPU/g干基质条件下批次酶解72h,酶解率达60%,葡萄糖浓度达83.4g/L。酸催化常压甘油有机溶剂酸预处理在明显缩短预处理时间的同时,能显著提高木质纤维素基质可酶解性,使后续工业化意义的浓醪酶解糖化成为可能。     

生物酶解法提取苦楝素工艺过程的研究

采用国产纤维素酶64001131,研究生物酶解法提取苦楝树皮中苦楝素的工艺条件。优化的工艺条件为:酶解液pH值4.5,酶解温度55℃,酶解时间1.5 h,酶浓度0.15 mg/mL。在此条件下,苦楝素的提取率达2.008%。     

非离子表面活性剂对木质纤维素酶解的促进作用

纤维素酶单位酶活力较低、酶用量较高及酶自身易失活等因素依然是木质纤维素工业生产能源和生物基产品的瓶颈性问题。本文尝试在木质纤维素基质水解时添加一些非离子型表面活性剂以减少纤维素酶用量,并对这些非离子型表面活性剂促进酶解效率提高的原因进行了初步探讨。研究发现,添加非离子性表面活性剂能提高木质纤维素的酶解,添加浓度为0.05 g/g底物,常压甘油自催化预处理麦草经过添加两种非离子表面活性剂Tween-80和PEG 6000后葡萄糖产量分别可提高20%左右;非离子表面活性剂对不含木质素的原料酶解产糖也有较大的提高,以滤纸为底物时葡萄糖产量提高近90%,以微晶纤维素为底物时分别提高70%以上;添加非离子表面活性剂使得酶解体系中扩散系数k升高,异相反应效率提高,酶促反应动力学Km值明显减小,显著提高底物对纤维素酶的亲和力。     

Palm date fibers: analysis and enzymatic hydrolysis

Waste palm dates were subjected to analysis for composition and enzymatic hydrolysis of their flesh fibers. The fruit contained 32% glucose and 30% fructose, while the water-insoluble fibers of its flesh consisted of 49.9% lignin and 20.9% polysaccharides. Water-insoluble fibers were settled to 55% of its initial volume in 12 h. The presence of skin and flesh colloidal fibers results in high viscosity and clogging problems during industrial processes. The settling velocity of the fibers was improved by enzymatic hydrolysis. Hydrolysis resulted in 84.3% conversion of the cellulosic part of the fibers as well as reducing the settling time to 10 minutes and the final settled volume to 4% of the initial volume. It implies easier separation of the fibers and facilitates fermentation processes in the corresponding industries. Two kinds of high- and low-lignin fibers were identified from the water-insoluble fibers. The high-lignin fibers (75% lignin) settled easily, while the low-lignin fibers (41.4% lignin) formed a slurry suspension which settled very slowly. The hydrophilicity of these low-lignin fibers is the major challenge of the industrial processes.     

酶解木质素的制备及其在聚氯乙烯改性中的应用

介绍了采用无机碱性水溶液和有机溶剂分离、提取酶解木质素的方法,研究了热塑性酶解木质素(FTPL10)对聚氯乙烯材料的力学性能、热稳定性的影响。结果表明:当FTPL10用量为5份时,改性聚氯乙烯材料的拉伸强度为30.66MPa,冲击强度为5.91kJ/m2,热稳定时间为2 091s,与未经酶解木质素改性的聚氯乙烯材料相比,拉伸强度略微下降,冲击强度提高了60%,热稳定时间延长了74%。     

半纤维素酶添加对碱处理甘蔗渣结构及酶解的影响

半纤维素作为木质纤维素的重要组分之一,通过氢键与纤维素的微纤丝结合,严重阻碍了纤维素表面与纤维素酶的接触,降低了酶解的效率。该试验以碱处理甘蔗渣作为底物,通过添加不同量的半纤维素酶去除不同比例的半纤维素。通过成分分析、X射线衍射(XRD)和扫描电镜(SEM)等手段分析添加半纤维素酶前后残渣的结构和酶解特性变化,发现随着半纤维素酶添加量的增大,残渣中木质素所占的比例逐渐增大,结晶指数逐渐增大,电镜表面沟壑逐渐加深,纤维束之间结构变得疏松。以半纤维素酶处理过的甘蔗渣作为底物,按照5FPU/g底物加入纤维素酶水解72h,与不添加半纤维素酶对照组相比,添加1600U/g半纤维素酶处理的试验组木聚糖的转化率提高了74.24%,葡聚糖转化率提高了35.30%。通过半纤维素酶添加可以有效促进纤维素酶解过程的进行,节约反应时间提高酶解转化率。     

竹青表面蜡质对纤维素酶解的影响研究进展

覆盖在竹子最表层的竹青蜡质层在竹子生长发育、适应外界环境方面起重要作用,同时也是阻碍生物炼制的主要抗降解屏障之一。目前,竹材的工业化加工利用大多以竹纤维为主,竹青和竹黄部分通常以加工剩余物的形式除去,不仅浪费资源,而且造成严重的环境污染。与不含蜡质层的竹肉和竹黄相比,含有蜡质层的竹青即使在预处理后其纤维素酶水解糖化效率仍十分低下。本文对竹青表皮蜡质的组成、蜡质成分对纤维素酶的相互作用以及预处理对蜡质的影响等方面研究进行综述,并探讨其存在的问题机理与研究前景。为提高以竹加工剩余物为原料的生物炼制效率提供理论依据和借鉴,为竹源天然蜡质提取及材料应用创新提供新途径。     

近临界水中鱼蛋白水解及水解液脱色研究

鱼类蛋白包括鱼肉及鱼类加工过程中产生的废弃物,文中采用近临界水技术,在反应温度180—320℃,反应压力5—26 MPa,反应时间5—60 min,无催化剂条件下将其制备成氨基酸,不但能提高附加值,而且有利于环境保护。实验结果发现,水解物含17种氨基酸,水解物中不同种类氨基酸的产率随反应温度、反应压力和反应时间的变化规律各不相同。对其中8种含量高、用途广、附加值高的氨基酸,分别得到了最佳水解工艺。以颗粒状活性炭为脱色剂,对水解液在不同pH值、活性炭用量、吸附温度和吸附时间条件下进行脱色实验,并考察脱色率和氨基酸损失率,结果表明,较好的脱色工艺条件为:pH=4.0,活性炭用量为0.020 g/mL,吸附温度45℃,吸附时间25 min,脱色率90%。被活性炭吸附的氨基酸,可以通过解吸附得到回收。     

厌氧水解的新解读：快速水解和慢速水解

挥发性脂肪酸（VFA）具有可生化性好、附加价值高等优点而得到广泛应用,厌氧消化产VFA是目前学术界的研究热点,但对有机物经过水解后的生物降解性能则少有论及,厌氧发酵过程中的水解产物存在“数量”和“质量”之间的权衡问题。为了进一步提高厌氧发酵过程中微生物对有机物的利用效率,突破水解对厌氧消化的限制性作用,从基质碳源释放快慢及厌氧发酵过程中碳源降解性能不同,本文将厌氧水解分为快速水解和慢速水解,分别阐述了两种水解方式的含义、分类以及优缺点,指出细胞内外碳源的释放速率和释放方式的不同是影响厌氧产酸和生物降解性能的决定性因素。最后指出快速水解与慢速水解相结合的分阶段联合处理方式,是今后厌氧消化产VFA的主要研究方向。     

High‐pressure enzymatic hydrolysis of oil

The hydrolysis of sunflower and soybean oil, catalyzed by two enzymes, non‐immobilized Candida rugosa and immobilized Candida antarctica lipase, was performed at atmospheric and high‐pressure. The results showed that at atmospheric pressure between 40 °C and 60 °C initial reaction rates were influenced by the temperature variation, as expected. Due to favorable physico‐chemical properties of dense gases as reaction media, hydrolysis of soybean oil was performed in non‐conventional solvents: in supercritical (SC) CO₂ and near‐critical propane. In SC CO₂ the activity of non‐immobilized Candida rugosa lipase decreased while the reaction rates of hydrolysis catalyzed by immobilized Candida antarctica lipase were 1.5‐fold higher than at atmospheric pressure. However, the reaction rates for the hydrolyses catalyzed by both lipases, were much higher in propane than at atmospheric pressure.     

Microscale recovery of total lipids from fish tissue by accelerated solvent extraction

A number of techniques are available for the extraction of lipids from a variety of tissues; however, conventional methods are characteristically labor intensive, typically involve large volumes of toxic solvents, and usually require at least 1 g of tissue. With the availability of accelerated solvent extraction (ASE) technology, the opportunity exists to modify classical lipid extraction techniques such that automated high-pressure, high-temperature extractions may be performed with the use of far smaller volumes of costly and harmsul solvents. Moreover, the high extraction efficiency attainable by ASE suggests that significantly less tissue would be required than is routinely used. This paper describes the adaptation of previously developed lipid extraction solvent systems for use with ASE toward the purpose of extracting total lipids from 100 mg of fish tissue. The efficacy of three solvent systems for lipid extraction from representative fish tissues, including a standard reference material, was explored using gravimetry and FA analysis by GC. A TG was used as a surrogate to monitor overall method performance. The findings herein demonstrate that microscale ASE represents an effective and efficient alternative to traditional lipid extraction techniques based on quantity and composition of extracted lipid, surrogate recovery, and precision.