Purification and Characteristics of Feruloyl Esterase from Aspergillus awamori G-2 Strain

ABSTRACT:  For food industry production processes and other uses, a mold that produces high levels of feruloyl esterase was obtained from laboratory mold collections and other sources. It was Aspergillus awamori G-2 that produces high levels of feruloyl esterase. The feruloyl esterase was purified using ion-exchange chromatography, size-exclusion chromatography, and HPLC chromatography. The enzyme was identified as a monomer protein using size-exclusion chromatography. Its optimum temperature and pH were, respectively, 40 °C and pH 5. Its activity was stable at pH 3 to 5. The enzyme was combined with xylan and starch, but it was absorbed by cellulose. The km of the feruloyl esterase was 0.0019% (0.01 mM). The enzyme showed stable activity at pH 3 and 50 °C, making this enzyme useful for food production.     

Utilization of feruloyl esterase and xylanase for the degradation of brewers＇ spent grain

橘青霉以麦糟为唯一碳源培养时,阿魏酸酯酶的最佳发酵时间为60 h,其酶活力可达40.8 mU/mL。在pH值为5.0、45 ℃、料液比1∶30（g∶mL）条件下,取47.5 U/mL的木聚糖酶粗酶液15 mL,加入1.0 g 麦糟的乙醇不溶物,反应12 h后,加入40.8 U/mL的阿魏酸酯酶粗酶液15 mL再反应12 h,阿魏酸和低聚木糖释放率分别为54.1%和161 mg/g （麦糟的乙醇不溶物）。实验结果还表明,阿魏酸酯酶与木聚糖酶存在协同作用,能极大提高麦糟中阿魏酸及低聚木糖的释放率,有利于麦糟的降解。     

阿魏酸酯酶与纤维素酶协同水解麦麸释放阿魏酸的研究

研究了重组阿魏酸酯酶和纤维素酶协同作用水解去淀粉麸皮对阿魏酸释放量的影响。以高效液相色谱法检测阿魏酸的提取量,结果显示,底物的超声预处理最佳条件为350 W,20 min,是未经超声处理的底物阿魏酸释放率的1.45倍,单独使用阿魏酸酯酶(re Ao Fae A)30 U时阿魏酸的释放率仅为4.1%,单独使用纤维素酶(r Au Cel12A)并不释放阿魏酸,当两种酶协同作用时,阿魏酸的释放率明显提高,经单因素试验确定双酶协同作用的最佳条件为:re Ao Fae A的最适添加量为30 U,r Au Cel12A的最适添加量为70 U,水解时间为10 h,水解温度为40℃,水解p H为5.0,料液质量体积比为1 g:30 m L,此时阿魏酸的释放率为23.6%。该结果表明,去淀粉麸皮中纤维素的降解,对提高阿魏酸酯酶水解释放阿魏酸效率具有重要作用。     

产阿魏酸酯酶菌株的筛选及其酚酸释放研究

本文采用平板筛菌和牛津杯透明圈试验法筛选具有产阿魏酸酯酶的菌株,将其接种于麸皮中进行固态发酵,采用分光光度法和HPLC法分析发酵麸皮其阿魏酸酯酶活力、总酚酸以及阿魏酸的释放量,并进行个体形态、菌落特征和TIS测序鉴定,结果表明:筛选出一株菌株的阿魏酸酯酶酶活优于其他菌株,在发酵第4 d该菌株的阿魏酸酯酶酶活达最高为163.5 m U/g;而且发酵麸皮释放总酚酸的能力较高,将麸皮的总酚酸量提高6.6倍,其中阿魏酸的量达到0.697%,比没有发酵的麸皮阿魏酸的含量增加了0.22%。对其进行分子鉴定,该菌株与烟曲霉(Aspergillus clavatus)同源性达100%,确认为烟曲霉命名为烟青。将该菌应用于麸皮发酵提高阿魏酸含量,是农副产品增值利用以及药用功能开发的主要研究方向。     

玉米麸皮中阿魏酰低聚糖的制备

以玉米麸皮为原料,利用纤维素酶辅助木聚糖酶制备阿魏酰低聚糖,通过正交试验对制备条件进行了优化,得到最佳制备条件：反应温度60℃,pH值5．0,反应时间36h,纤维素酶质量浓度8g／L,木聚糖酶质量浓度6g／L,底物质量浓度165g／L,在此条件下,产物中阿魏酰低聚糖的浓度达到2．127mmol／L．     

Exposure or release of ferulic acid from wheat aleurone: Impact on its antioxidant capacity

The relationship between the aleurone cell integrity and the exposure or release of bioavailable ferulic acid (FA) with the antioxidant capacity of aleurone in in vitro and under simulated gastric conditions was explored. The antioxidant capacity of aleurone was increased by around 2-fold when its median particle size was reduced to under 50 μm. The opening of aleurone cells increased the physical exposure of FA bound to the insoluble polysaccharides, which seemed to be responsible of the increased antioxidant capacity. Synergistic combination of xylanase and feruloyl esterase was found to be the most efficient enzymatic treatment releasing up to 86% of total FA in bioaccessible forms. This enzymatic treatment significantly enhanced the radical scavenging activity of aleurone by up to 4-fold, which overlapped the overall antioxidant potential estimated from the total content of FA in aleurone. The improvement in the antioxidant capacity of aleurone was also observed in the simulated gastric digestion by inhibition of lipid oxidation.     

阿魏酸酯酶功能特性及应用概述

阿魏酸酯酶(ferulic acid esterase,FAE)广泛存在于自然界,属于羧酸酯水解酶的一个亚类。经过一系列反应,其能将天然半纤维素和果胶中的酚酸、阿魏酸(ferulic acid,FA)及对香豆酸等释放出来,是一种具有应用潜力的酶。因此,各行业通过相关科学技术来利用FAE,以获得如人畜食品、纸浆和纸张、燃料乙醇和药物制剂等产品。该文对阿魏酸酯酶的酶学特性、生产研究、结构解析及生物技术中的潜在应用等方面进行综述,旨在工业需求的推动下,加快阿魏酸酯酶的科学研究发现,进行知识转移,对该类物质实现高效利用,并致力于植物生物质转化和生产高附加值分子的生物合成。     

泡盛曲霉发酵麦麸过程中酚类物质含量的变化与三种酶活性的相关性

本研究以小麦麦麸为原料,分别采用3株泡盛曲霉(分别为GIM3.266,GIM3.4,GIM3.5)进行连续8天的固态发酵,每天分别提取发酵过程中小麦麦麸中不同存在形态酚类物质并测定其含量,同时,每天测定3株泡盛曲霉在发酵过程中木聚糖酶、纤维素酶以及阿魏酸酯酶的活性变化。结果表明,与未经发酵的麦麸相比,经泡盛曲霉发酵后的麦麸游离型酚类物质的含量显著增加,总酚含量可高达11561.30μg/g麸皮,比发酵前的麸皮总酚提高了169%;另外,酶活力的结果表明:纤维素酶活力在发酵前三天显著增大,其活力最高可达156.70 U,木聚糖酶活力随总酚释放量的增加而增加,最高可达34123.00 U,阿魏酸酯酶活力在发酵中后期达到顶峰,最高可达2642.60 U。总体而言,麦麸发酵过程中酚类物质的释放量与木聚糖酶和阿魏酸酯酶的酶活力呈正相关,说明发酵后麦麸中酚类物质提取量的显著增加与这两种酶密切相关,这两种酶活力高的泡盛曲霉菌株,更有利于释放麦麸中的酚类物质。