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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。 相似文献
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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。 相似文献
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为提高白酒酿造过程中阿魏酸的含量,该研究从浓香型白酒大曲中分离筛选产阿魏酸酯酶的菌株,通过形态观察、生理生化试验及分子生物学技术对其进行菌种鉴定,并以阿魏酸酯酶活力为评价指标,采用单因素试验及响应面试验对其发酵条件进行优化。结果表明,筛选得到一株产阿魏酸酯酶的菌株,编号为M2。经鉴定,其为一株蜡样芽孢杆菌(Bacillus cereus),该菌株产阿魏酸酯酶的最佳发酵条件为接种量5%,发酵温度30℃,初始pH值5.5,发酵时间72 h。在此优化条件下,阿魏酸酯酶活力为33.89 U/L,比优化前提高10.03%,具有较强的产阿魏酸酯酶能力。 相似文献
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筛选高产阿魏酸酯酶菌株,并对其发酵特性及在黄酒中的应用进行研究。通过初筛及复筛,根据菌落形态、生理生化特征及rDNA ITS1-5.8S-ITS2基因序列的系统发育树构建分析进行菌株鉴定;以阿魏酸含量为衡量指标,考察温度、初始pH、麦麸添加量、酒度等因素对菌株发酵过程中产阿魏酸的影响;菌株经过富集培养反添加到黄酒酿造过程中。筛选出一株高产阿魏酸酯酶霉菌,经分子鉴定为枝状枝孢霉菌株(Cladosporium cladosporioides),保藏在中国典型培养物保藏中心,保藏编号为CCTCC NO:M2015549。菌株在30℃、pH 4.8、麦麸添加量5%、酒度9%条件下培养3 d,菌株产阿魏酸酯酶活力最高可达175.4 U/L。当菌株添加量为0.010%时,黄酒阿魏酸含量提高到150%。筛选到一株新的高产阿魏酸酯酶的枝状枝孢霉菌株,并将其反添加到黄酒酿造过程中,对黄酒阿魏酸含量的提高起到积极作用,证明菌株在产阿魏酸酯酶及富含阿魏酸功能性食品开发方面具有很大潜力。 相似文献
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高产阿魏酸酯酶菌株的筛选及其固态发酵的研究 总被引:4,自引:0,他引:4
从自然界中采集土壤样本,使用阿魏酸乙酯为唯一碳源的选择性培养基;进行初筛、固态发酵产酶试验、复筛得到1株高产阿魏酸酯酶的菌株,通过形态学观察鉴定为黑曲霉。对该菌株固态发酵产阿魏酸酯酶的培养条件进行了研究,单因素实验结果表明,相同汽爆条件下汽爆稻草的发酵产酶酶活较高;在固液比1∶3、初始pH3、麸皮添加量25%、30℃下发酵3d后酶活最高,可达445.1mU/g。 相似文献
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为了评价两种丝状真菌对小麦麸皮降解能力,以及对酚酸释放能力的差异,本实验在不同时间进行总酚和9种酚酸组分的变化监测并且结合阿魏酸酯酶和木聚糖酶活性,电镜观察小麦麸皮表面结构对黑曲霉和棒曲霉进行综合评估。黑曲霉发酵7 d阿魏酸含量达到416.56μg/g麦麸。次之是没食子酸,从7.71μg/g麦麸增加到105.77μg/g麦麸。棒曲霉阿魏酸含量达到200.81μg/g麦麸。次之是丁香酸,没食子酸,对羟基苯甲酸和香豆酸,从分别从原麦麸中含量14.7μg/g麦麸、7.71μg/g麦麸、8.57μg/g麦麸和5.62μg/g麦麸增长至88.4μg/g麦麸,60.38μg/g麦麸,55.56μg/g麦麸,52.93μg/g麦麸。黑曲霉对麦麸的降解能力及对阿魏酸和没食子酸的释放能力都显著优于棒曲霉,而对丁香酸的释放能力略差于棒曲霉。电镜结果显示,黑曲霉降解能力优于棒曲霉。 相似文献
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A partial purification of ferulic acid esterase, which degrades feruloyl glycerol, has been achieved from barley malt in small yields. Coloured and viscous materials were removed from the malt extract using batch‐elution anion exchange chromatography. Further steps included gradient elution anion exchange chromatography and gel filtration chromatography. Estimations of the molecular weight varied greatly from 22KDa to 158 KDa, possibly because the protein interacted with the matrix of the gel exclusion chromatography column and because multiple forms of the enzyme were present. The partially purified feruloyl esterase had an apparent Km of 0.46% feruloyl glycerol. However more than one enzyme may be present and the substrate contains two isomers and so Michelis‐Menten kinetics may not be appropriate. 相似文献
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本研究以黑曲霉(Aspergillus niger)作菌种,采用液体深层发酵法制备出含有阿魏酸酯酶(FAE)和阿拉伯木聚糖酶(AX)的混合酶制剂,探讨了它们协同作用麦麸制备阿魏酸和低聚糖的工艺条件.结果表明,FAE和AX协同作用的最适反应pH为4.4,最适反应温度为50℃.采用混合酶制剂作用于去淀粉麦麸(DSWB)发现,通过3次降解后,麦麸降解率达55.46%,大大高于文献报道的、采用单一酶作用的结果.扫描电子显微镜(SEM)观察发现,DSWB在酶处理前后的微观结构发生了显著变化. 相似文献
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Ferulic acid esterase activity was detected in extracts of barley malt using an assay employing a novel artificial substrate, mono‐feruloyl glycerol. Mono‐feruloyl glycerol has been synthesized and analysed to determine its degree of substitution and purity. It consists of a mixture of the two isomers 1‐feruloyl glycerol and 2‐feruloyl glycerol. The extraction of ferulic acid esterase and its assay conditions have been optimised. The presence of both a detergent and reduced glutathione in the extraction medium increased the amount of enzyme extracted. A pH of 7.5 was optimal for enzyme activity. The enzyme in solution was only stable up to 30°C. The crude extract containing the enzyme released free ferulic acid from both soluble and insoluble cell wall materials. After extraction of the soluble enzyme, insoluble enzyme, capable of releasing free ferulic acid from feruloyl glycerol, was detected in the residual grain solids. 相似文献
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Dominik Szwajgier 《Journal of the Institute of Brewing》2011,117(3):427-434
A purified extracellular ferulic acid esterase from Lactobacillus acidophilus K1 was successfully used during mashing for the release of free phenolic acids into sweet wort. The enzyme was produced in bioreactors and partially purified to obtain the monoenzyme preparation. Release of free ferulic and vanillic acid into the wort at 52°C (with the use of 4.09–14.60 enzyme activity units/L of the mash) and ferulic acid at 62°C (14.60 units/L) was observed. Free p‐OH‐benzoic and syringic acids were effectively released at 26°C at each enzyme concentration used. Free p‐OH‐benzoic acid was also released by the enzyme (14.60 U/L) at 52°C‐74°C. Free protocatechuic acid was effectively hydrolyzed by the enzyme preparation (8.75 U/L and 14.60 U/L) at 26°C‐52°C. Free caffeic acid (effectively released at 26°C‐62°C) originated from chlorogenic acid. No p‐coumaric acid was released due to the action of bacterial esterase during mashing. Ferulic acid esterase from L. acidophilus K1 exhibited no ability to release free phenolic acids during mashing at 62°C or at 74°C due to its low thermostability. In conclusion, L. acidophilus K1 is an attractive source for the production of ferulic acid esterase, which can be useful for the release of antioxidant phenolic acids in the early stages of mashing. 相似文献