Enzymatic hydrolysis

酶水解
  • 文章类型: Journal Article
    酶固定化的使用在饮料加工中变得越来越流行,由于这种方法具有显著的优势,如增强酶性能和扩大应用,同时允许通过简单的过滤过程容易终止。这篇文献综述分析了大约120篇文章,2000年至2023年在WebofScience上发表,重点研究了用于饮料加工应用的酶固定系统。固定化对酶活性的影响,包括对化学和动力学性质的影响,可回收性,在连续过程中的可行性,进行了评估。这些系统在饮料生产中的应用,比如葡萄酒,啤酒,果汁,牛奶,和植物性饮料,进行了检查。固定化过程有效地提高了pH值和热稳定性,但通过降低最大速度和米氏-门顿常数对动力学特性产生了负面影响。然而,它允许多次重复使用,并促进连续流动过程。封装还允许通过简单过滤简化从饮料中去除酶的过程控制。不需要昂贵的热处理,这可能会导致产品质量损失。
    The use of enzyme immobilisation is becoming increasingly popular in beverage processing, as this method offers significant advantages, such as enhanced enzyme performance and expanded applications, while allowing for easy process termination via simple filtration. This literature review analysed approximately 120 articles, published on the Web of Science between 2000 and 2023, focused on enzyme immobilisation systems for beverage processing applications. The impact of immobilisation on enzymatic activity, including the effects on the chemical and kinetic properties, recyclability, and feasibility in continuous processes, was evaluated. Applications of these systems to beverage production, such as wine, beer, fruit juices, milk, and plant-based beverages, were examined. The immobilisation process effectively enhanced the pH and thermal stability but caused negative impacts on the kinetic properties by reducing the maximum velocity and Michaelis-Menten constant. However, it allowed for multiple reuses and facilitated continuous flow processes. The encapsulation also allowed for easy process control by simplifying the removal of the enzymes from the beverages via simple filtration, negating the need for expensive heat treatments, which could result in product quality losses.
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  • 文章类型: Journal Article
    大量研究表明,不同硅酸盐细菌对矿石的共浸出显着提高了生物浸出系统的性能。然而,不同硅酸盐细菌协同或互补增强含锂硅酸盐浸出过程的机理尚不清楚。本研究讨论了两种代谢不同的硅酸盐细菌的组合存在对锂云母的浸出影响,目的是理解在浸出过程中由于粘液芽孢杆菌和环状芽孢杆菌的存在而产生的协同作用。结果表明,细菌分泌的多糖和蛋白质含有官能团,如-OH和-COOH,在复杂的矿石分解中起着重要作用。有机酸起着酸蚀和络合作用。粘液芽孢杆菌和环状芽孢杆菌表现出较低的个体浸出效率,主要是由于它们的有机酸分泌弱。此外,粘液芽孢杆菌生产的多产多糖导致细菌聚集,与矿物的接触能力减弱。环状芽孢杆菌在共浸出过程中通过酶水解分解了胶质芽孢杆菌产生的过量多糖,为这两种菌株提供后期营养供应。两个菌株的共生增强了两个菌株的合成和代谢能力,导致有机酸分泌增加。此外,胶质芽孢杆菌产生的蛋白质和腐殖酸增强,集体提高浸出效率。这些发现表明,不同菌株在浸出过程中分泌的主要代谢产物不同。共浸出过程中生物浸出效率的提高归因于它们的有效协同代谢。这项工作有助于构建高效的工程微生物群落,以提高硅酸盐矿物的浸出效率,并揭示了微生物共培养提高生物浸出的可行性。
    Numerous studies have demonstrated that the co-leaching of ores by different silicate bacteria significantly improves the performance of bioleaching systems. Nevertheless, the mechanism of different silicate bacteria synergistically or complementarily enhanced the leaching process of lithium-containing silicate remains unclear. This study discussed the leaching impact of the combined presence of two metabolically distinct silicate bacteria on lepidolite, with the aim of comprehending the synergistic effect resulting from the presence of Bacillus mucilaginosus and Bacillus circulans in the leaching process. The results indicated that the polysaccharides and proteins secreted by bacteria-containing functional groups such as -OH and -COOH, which played an important role in the complex decomposition of ores. Organic acids played the role of acid etching and complexation. Bacillus mucilaginosus and Bacillus circulans exhibited low individual leaching efficiency, primarily due to their weak organic acid secretion. Moreover, the prolific polysaccharide production by Bacillus mucilaginosus led to bacterial aggregation, diminishing contact capability with minerals. Bacillus circulans decomposed the excessive polysaccharides produced by Bacillus mucilaginosus through enzymatic hydrolysis in the co-bioleaching process, providing later nutrient supply for both strains. The symbiosis of the two strains enhanced the synthesis and metabolic capabilities of both strains, resulting in increased organic acid secretion. In addition, protein and humic acid production by Bacillus mucilaginosus intensified, collectively enhancing the leaching efficiency. These findings suggested that the primary metabolic products secreted by different bacterial strains in the leaching process differ. The improvement in bioleaching efficiency during co-leaching was attributed to their effective synergistic metabolism. This work contributes to the construction of an efficient engineering microbial community to improve the efficiency of silicate mineral leaching, and reveals the feasibility of microbial co-culture to improve bioleaching.
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  • 文章类型: Journal Article
    Ovolucin(OM),具有不溶部分的是粘性糖蛋白,在鸡蛋白蛋白中发现。OM的酶解产物具有水溶性和生物活性。本研究调查了通过使用各种蛋白水解酶(Alcalase®,菠萝蛋白酶,α-胰凝乳蛋白酶,Neutrase®,胰酶,木瓜蛋白酶,Protamax®,和胰蛋白酶)在RAW264.7细胞中。结果表明,用胰酶(OMPA)制备的OMH在RAW264.7细胞中产生的亚硝酸盐氧化物水平最高,通过上调诱导型一氧化氮合酶mRNA表达。促炎细胞因子如肿瘤坏死因子-α和白细胞介素-6的产生随着细胞因子mRNA的表达而增加。OMPA对丝裂原活化蛋白激酶信号通路的影响是p38,c-JunNH2末端激酶磷酸化增加,和细胞外信号调节激酶以浓度依赖的方式。因此,OMPA可以在功能性食品工业中用作潜在的免疫刺激剂。
    Ovomucin (OM), which has insoluble fractions is a viscous glycoprotein, found in egg albumin. Enzymatic hydrolysates of OM have water solubility and bioactive properties. This study investigated that the immunostimulatory effects of OM hydrolysates (OMHs) obtained by using various proteolytic enzymes (Alcalase®, bromelain, α-chymotrypsin, Neutrase®, pancreatin, papain, Protamax®, and trypsin) in RAW 264.7 cells. The results showed that OMH prepared with pancreatin (OMPA) produced the highest levels of nitrite oxide in RAW 264.7 cells, through upregulation of inducible nitric oxide synthase mRNA expression. The production of pro-inflammatory cytokines such as tumor necrosis factor-α and interleukin-6 were increased with the cytokines mRNA expression. The effect of OMPA on mitogen-activated protein kinase signaling pathway was increased the phosphorylation of p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase in a concentration-dependent manner. Therefore, OMPA could be used as a potential immune-stimulating agent in the functional food industry.
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  • 文章类型: Journal Article
    尽管现代食品卫生取得了进步,由于微生物污染引起的食物中毒仍然是一个全球性问题,对人类健康构成了极大的威胁。尤其是,单核细胞增生李斯特菌和金黄色葡萄球菌是在具有生物膜的食物接触表面上发现的革兰氏阳性细菌。这些食源性病原体每年引起相当数量的食物中毒和感染。Ovomucin(OM)是蛋白中的水不溶性凝胶型糖蛋白。酶水解可用于提高OM的生物活性。本研究旨在调查是否使用五种商业酶(Alcalase®,菠萝蛋白酶,α-胰凝乳蛋白酶,木瓜蛋白酶,和胰酶)可以抑制单核细胞增生李斯特菌ATCC15313,单核细胞增生李斯特菌H7962,金黄色葡萄球菌KCCM11593和金黄色葡萄球菌7的生物膜形成。特别是,用木瓜蛋白酶(OMPP;500μg/mL)制备的OMH显着抑制了单核细胞增生李斯特菌ATCC15313,单核细胞增生李斯特菌H7962,金黄色葡萄球菌KCCM11593和金黄色葡萄球菌7的生物膜形成,80.28%,91.70%,79.00%,分别。此外,OMPP降低了代谢活性,胞外多糖生产(EPS),粘附能力,以及与这些细菌菌株的生物膜形成相关的基因表达。这些结果表明,OMH,尤其是OMPP,对单核细胞增生李斯特菌和金黄色葡萄球菌发挥抗生物膜作用。因此,OMPP可作为天然抗生物膜剂用于控制食品工业中的食物中毒。
    Despite modern advances in food hygiene, food poisoning due to microbial contamination remains a global problem, and poses a great threat to human health. Especially, Listeria monocytogenes and Staphylococcus aureus are gram-positive bacteria found on food-contact surfaces with biofilms. These foodborne pathogens cause a considerable number of food poisoning and infections annually. Ovomucin (OM) is a water-insoluble gel-type glycoprotein in egg whites. Enzymatic hydrolysis can be used to improve the bioactive properties of OM. This study aimed to investigate whether ovomucin hydrolysates (OMHs) produced using five commercial enzymes (Alcalase®, Bromelain, α-Chymotrypsin, Papain, and Pancreatin) can inhibit the biofilm formation of L. monocytogenes ATCC 15313, L. monocytogenes H7962, S. aureus KCCM 11593, and S. aureus 7. Particularly, OMH prepared with papain (OMPP; 500 μg/mL) significantly inhibited biofilm formation in L. monocytogenes ATCC 15313, L. monocytogenes H7962, S. aureus KCCM 11593, and S. aureus 7 by 85.56 %, 80.28 %, 91.70 %, and 79.00 %, respectively. In addition, OMPP reduced the metabolic activity, exopolysaccharide production (EPS), adhesion ability, and gene expression associated with the biofilm formation of these bacterial strains. These results suggest that OMH, especially OMPP, exerts anti-biofilm effects against L. monocytogenes and S. aureus. Therefore, OMPP can be used as a natural anti-biofilm agent to control food poisoning in the food industry.
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  • 文章类型: Journal Article
    在这项研究中,目的是研究一步法从einkorn面粉中直接释放BAPs的情况。因此,取消了蛋白质提取步骤,从而降低加工成本。商业蛋白酶(Alcalase,调味剂,中和酶,和胰蛋白酶),和来自莫哈韦芽孢杆菌EBTA7的粗酶用于水解einkorn面粉(30%,w/v)50-60°C的溶液离心后的上清液用于生物活性和技术功能性测试。所有水解产物都表现出显著的抗氧化能力,DPPH的值范围为17.7至33.0μmolTE/g,对于ABTS,107至190μmolTE/g,和0.09至3.08mgEDTA/g的离子螯合活性。Alcalase和Flavourzyme水解具有最高的DPPH活性,而莫雅芽孢杆菌EBTA7酶具有相对较高的ABTS和离子螯合活性。值得注意的是,莫雅芽孢杆菌。EBTA7粗酶水解物表现出较高的吸油能力(2.94g油/g水解物),强效乳液(227分钟),和泡沫稳定性(94%)与商业酶相比。FTIR光谱证实了肽的二级结构的变化。所有水解产物均表现出负ζ电位。SDS-PAGE显示的分子量范围为14至70kDa,受酶类型和水解程度的影响。总的来说,莫雅芽孢杆菌。EBTA7水解产物显示出相当大的生物和技术功能特性。
    In this study, it was aimed to investigate the direct release of BAPs from einkorn flour in one-step process. Thus, the protein extraction step was eliminated, thereby reducing processing cost. Commercial proteases (Alcalase, Flavourzyme, Neutrase, and Trypsin), and crude enzyme from Bacillus mojavensis sp. EBTA7 were used for hydrolyzing einkorn flour (30 %, w/v) solutions at 50-60 °C. The supernatants after centrifugation were used for bioactivity and techno-functionality tests. All hydrolysates demonstrated significant antioxidant capacities, with values ranging from 17.7 to 33.0 μmol TE/g for DPPH, 107 to 190 μmol TE/g for ABTS, and 0.09 to 3.08 mg EDTA/g for ion-chelating activities. Alcalase and Flavourzyme hydrolysis had the highest DPPH activities, while Bacillus mojavensis sp. EBTA7 enzyme yielded relatively high ABTS and ion-chelating activities. Notably, Bacillus mojavensis sp. EBTA7 crude enzyme hydrolysates demonstrated higher oil absorption capacity (2.94 g oil/g hydrolysate), robust emulsion (227 min), and foam stability (94 %) compared to commercial enzymes. FTIR spectroscopy confirmed variations in the secondary structure of peptides. All hydrolysates exhibited negative zeta potentials. The SDS-PAGE showcased MW ranged from 14 to 70 kDa, which was influenced by both the enzyme type and the degree of hydrolysis. Overall, Bacillus mojavensis sp. EBTA7 hydrolysates revealed considerable bio and techno-functional characteristics.
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  • 文章类型: Journal Article
    精油显示出几种生物学特性,如抗菌活性,但对其可用性和稳定性有限制。为了最大限度地提高其抗菌效果和对环境条件的保护,Pickering型乳液用于以纤维素纳米纤维(CNF)为乳液稳定剂的牛至精油(OEO)。酶水解用于从食品工业废物(木薯皮)生产CNF,获得环境可持续的乳液稳定剂。评价了纳米纤维的不同性质如何影响乳液的稳定性。此外,鉴于在可生物降解的活性涂料中的目标应用,分散相的组成有所不同(OEO和葵花籽油-SO的比例不同)。即使在非常低的浓度(0.01%w/w),CNF能够使用油混合物(OEO+SO)形成具有小液滴尺寸的动力学稳定乳液。稳定机制不是纯粹的皮克林,因为界面张力降低了。对细菌和真菌链格孢菌观察到优异的抗菌活性,证明了将这些乳液应用于活性体系如涂层和薄膜的能力。当使用油的混合物时,观察到乳液稳定性的改善。考虑到热处理的成本和稳定性,这是非常有利的,因为在最终应用中保持了所需的抗微生物活性。
    Essential oils show several biological properties, such as antimicrobial activity, but have limitations regarding their availability and stability. To maximize their antimicrobial effect and protection against environmental conditions, Pickering-type emulsions were used to vehiculate oregano essential oil (OEO) using cellulose nanofibers (CNF) as emulsion stabilizer. Enzymatic hydrolysis was used to produce CNF from a food industry waste (cassava peel), obtaining an environmentally sustainable emulsion stabilizer. It was evaluated how the different properties of the nanofibers affected the stability of the emulsions. Furthermore, the composition of the dispersed phase was varied (different ratios of OEO and sunflower oil-SO) in view of the target application in biodegradable active coatings. Even at very low concentration (0.01 % w/w), CNF was able to form kinetically stable emulsions with small droplet sizes using oil mixtures (OEO + SO). The stabilization mechanism was not purely Pickering, as there was a reduction in interfacial tension. Excellent antimicrobial activity was observed against bacteria and the fungus Alternaria alternata, demonstrating the ability to apply these emulsions in active systems such as coatings and films. An improvement in the stability of emulsions was observed when using a mixture of oils, which is extremely advantageous considering costs and stability to heat treatments, since the desired antimicrobial activity is maintained for the final application.
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  • 文章类型: Journal Article
    开发了一种由氯化胆碱(ChCl)基低共熔溶剂(DES)和甲基异丁基酮(MIBK)组成的简便两相体系,以实现糠醛的生产。一锅法从桉树中分离木质素并制备可发酵葡萄糖。结果表明,ChCl/1,2-丙二醇/MIBK体系具有将半纤维素转化为糠醛的最佳性能。在最佳条件下(MRChCl:1,2-丙二醇=1:2,原料:DES:MIBK比例=1:4:8g/g/mL,0.075mol/LAlCl3·6H2O,140°C,和90分钟),糠醛产率和葡萄糖产率分别达到65.0和92.2%,分别。同时,低分子量木质素(1250-1930g/mol),低多分散性(DM=1.25-1.53)和高纯度(碳水化合物含量仅为0.08-2.59%)从两相系统中再生。随着预处理温度的升高,再生木质素中的β-O-4、β-β和β-5键逐渐断裂,酚羟基的含量增加,但脂肪族羟基的含量下降。该研究为生物炼制过程中木质纤维素的综合利用提供了新的策略。
    A facile biphasic system composed of choline chloride (ChCl)-based deep eutectic solvent (DES) and methyl isobutyl ketone (MIBK) was developed to realize the furfural production, lignin separation and preparation of fermentable glucose from Eucalyptus in one-pot. Results showed that the ChCl/1,2-propanediol/MIBK system owned the best property to convert hemicelluloses into furfural. Under the optimal conditions (MRChCl:1,2-propanediol = 1:2, raw materials:DES:MIBK ratio = 1:4:8 g/g/mL, 0.075 mol/L AlCl3·6H2O, 140 °C, and 90 min), the furfural yield and glucose yield reached 65.0 and 92.2 %, respectively. Meanwhile, the lignin with low molecular weight (1250-1930 g/mol), low polydispersity (DM = 1.25-1.53) and high purity (only 0.08-2.59 % carbohydrate content) was regenerated from the biphasic system. With the increase of pretreatment temperature, the β-O-4, β-β and β-5 linkages in the regenerated lignin were gradually broken, and the content of phenolic hydroxyl groups increased, but the content of aliphatic hydroxyl groups decreased. This research provides a new strategy for the comprehensive utilization of lignocellulose in biorefinery process.
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  • 文章类型: Journal Article
    木质纤维素是化石燃料的有希望的替代品。监测木质纤维素颗粒的质量和尺寸变化而不破坏过程可以帮助调整预处理和酶促水解。传统的筛分方法不足。开发了一种利用聚焦光束反射率测量(FBRM)的方法,以建立FBRM弦信息(弦长和计数)与通过筛分量化的颗粒特征(重量和尺寸)之间的数学相关性。结果表明,颗粒尺寸与平方加权的中值弦长(Lsqr)呈线性相关,R2为0.93。Further,可以使用Lsqr和弦计数(R20.98)预测实时散装颗粒质量。这些相关性适用于53μm至358.5μm的范围。玉米秸秆酶解的实时监测证明了FBRM的实际适用性。本研究介绍了一种在线表征木质纤维素颗粒的新方法,从而增强木质纤维素生物精炼厂。
    Lignocellulose presents a promising alternative to fossil fuels. Monitoring the mass and size changes of lignocellulosic particles without disrupting the process can assist in adjusting pretreatment and enzymatic hydrolysis, where conventional sieving methods fall short. A method utilizing focused beam reflectance measurement (FBRM) was developed to establish mathematical correlations between FBRM chord information (chord length and count) and particle characteristics (weight and size) quantified through sieving. Results indicate particle size exhibits a linear correlation with the square weighted median chord length (Lsqr) with R2 at 0.93. Further, real-time bulk particle mass can be predicted using Lsqr and chord count (R2 0.98). These correlations are applicable in range 53 μm to 358.5 μm. Real-time monitoring of enzymatic hydrolysis of corn stalks has demonstrated the practical applicability of FBRM. This study introduces a novel approach for online characterization of lignocellulosic particles, thereby enhancing lignocellulosic biorefineries.
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  • 文章类型: Journal Article
    这项工作的目的是发展,第一次,可持续战略,基于超声辅助提取的使用,天然深共晶溶剂,和加压液体萃取,从石灰(柑橘xlatifolia)果皮中提取蛋白质,并评估其释放生物活性肽的潜力。PLE显示最大的蛋白质提取(66-69%),使用三种不同的酶水解(Alcalase2.4LFG,Alcalase®PURE2.4L,和嗜热菌蛋白酶)。评价了释放肽的体外抗氧化和降压活性。尽管所有水解产物都显示出抗氧化和降压活性,用嗜热菌蛋白酶获得的水解产物显示出最显著的值。由于所有水解产物中的总酚类含量较低,肽可能是这些生物活性的主要贡献者。通过UHPLC-QTOF-MS分析水解产物,鉴定出总共98种不同的肽。这些肽中的大多数富含与抗氧化活性相关的氨基酸。
    The aim of this work was to develop, for the first time, sustainable strategies, based on the use of Ultrasound-Assisted Extraction, Natural Deep Eutectic Solvents, and Pressurized Liquid Extraction, to extract proteins from lime (Citrus x latifolia) peels and to evaluate their potential to release bioactive peptides. PLE showed the largest extraction of proteins (66-69%), which were hydrolysed using three different enzymes (Alcalase 2.4 L FG, Alcalase®PURE 2.4 L, and Thermolysin). The in vitro antioxidant and antihypertensive activities of released peptides were evaluated. Although all hydrolysates showed antioxidant and antihypertensive activity, the hydrolysate obtained with Thermolysin showed the most significant values. Since the Total Phenolic Content in all hydrolysates was low, peptides were likely the main contributors to these bioactivities. Hydrolysates were analyzed by UHPLC-QTOF-MS and a total of 98 different peptides were identified. Most of these peptides were rich in amino acids associated with antioxidant activity.
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  • 文章类型: Journal Article
    商品大豆蛋白粉(SPI)的低复水特性,一种主要的植物性食品成分,限制了植物性食品的发展。本研究提出了一种大豆卵磷脂改性与Alcalase水解相结合的处理方法,以改善大豆蛋白粉的复水,以及其他加工性能(乳化,粘度)。结果表明,大豆蛋白-大豆卵磷脂复合粉,水解30分钟(SPH-SL-30),具有最小的颗粒尺寸,最小的zeta电位,最高的表面疏水性,和均匀的微观结构。此外,在SPH-SL-30中,α-螺旋结构/β-折叠结构的比值最小。在测量复水特性后,乳化性能,和粘度,发现SPH-SL-30具有3.04min的最短润湿时间,最短的分散时间为12.29s,最高溶解度为93.17%,乳化活性最高,为32.42m2/g,最高乳化稳定性98.33min,最低粘度为0.98Pa.s.这表明大豆卵磷脂改性与碱性蛋白酶水解相结合的处理破坏了大豆蛋白的结构,改变了它的物理化学性质,并提高其功能性能。在这项研究中,采用大豆卵磷脂改性与Alcalase水解相结合的方法对大豆蛋白进行改性,以改善大豆蛋白粉的加工特性,为其在植物性食品领域的高值化利用提供理论依据。
    The low rehydration properties of commercial soy protein powder (SPI), a major plant-based food ingredient, have limited the development of plant-based foods. The present study proposes a treatment of soy lecithin modification combined with Alcalase hydrolysis to improve the rehydration of soy protein powder, as well as other processing properties (emulsification, viscosity). The results show that the soy protein-soy lecithin complex powder, which is hydrolyzed for 30 min (SPH-SL-30), has the smallest particle size, the smallest zeta potential, the highest surface hydrophobicity, and a uniform microstructure. In addition, the value of the ratio of the α-helical structure/β-folded structure was the smallest in the SPH-SL-30. After measuring the rehydration properties, emulsification properties, and viscosity, it was found that the SPH-SL-30 has the shortest wetting time of 3.04 min, the shortest dispersion time of 12.29 s, the highest solubility of 93.17%, the highest emulsifying activity of 32.42 m2/g, the highest emulsifying stability of 98.33 min, and the lowest viscosity of 0.98 pa.s. This indicates that the treatment of soy lecithin modification combined with Alcalase hydrolysis destroys the structure of soy protein, changes its physicochemical properties, and improves its functional properties. In this study, soy protein was modified by the treatment of soy lecithin modification combined with Alcalase hydrolysis to improve the processing characteristics of soy protein powders and to provide a theoretical basis for its high-value utilization in the plant-based food field.
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