Protein Extraction from Heat-stabilized Defatted Rice Bran: II. The Role of Amylase, Celluclast, and Viscozyme

ABSTRACT: The effectiveness of 3 carbohydrases for protein extraction from heat-stabilized defatted rice bran (HDRB) was evaluated. Amylase, viscozyme and celluclast extracted a maximum of 45.4, 12.1, and 28.5% protein, respectively. Further study showed that extracted protein ranged from 9.5 to 58.4% under conditions of water to bran ratio (5:1 to 20:1), α-amylase (0 to 110000 units/10 g rice bran), temperature (35 to 55 °C), and time (1 to 8 h). The maximum protein extracted was 58.4% with a water to bran ratio of 17:1, 87637 units amylase, and 50.9 °C. These results suggest that impure food-grade amylase containing protease is more effective than celluclast and viscozyme in protein extraction from HDRB.     

纤维素酶分子的计算机辅助序列分析

利用生物大分子数据库（EMBL、SWALL和PDB）及软件技术（DNASIS和AntheProt）对嗜热子囊菌（Thermoascus aurantiacus)的产生的一种纤维素酶分子进行了序列分析及结构预测；通过多种来源的纤维素酶的比较研究，分析了其催化区、活性位点及局部保守序列。     

Improved cellulase production by stable Clostridium cellulolyticum mutants

Methods for the production of mutants of the cellulase producer Clostridium cellulolyticum ATCC 35319 were examined using an agar plate screening technique. Spontaneous and UV light-induced mutants were isolated, some of which exhibited a high level of both endoglucanase and exoglucanase activities as assayed using CMC (carboxymethyl cellulose) and PNPCb (paranitrophenyi-β-cellobioside) respectively. The volumetric enzyme activities were up to 2.5 times those of the wild strain.     

丝状真菌纤维素酶合成机制的研究

丝状真菌产生的纤维素酶被认为是最有应用前景的,但就目前酶生产效率来看,离实际应用还有很大的差距,需要对酶的合成调节机制有更为全面和深刻的了解。纤维素酶的生物合成受诱导和阻遏双重控制,酶的生产既有赖于低廉的保持一定浓度诱导物的存在,又必须清除分解代谢产物对酶合成的阻遏,其中的详细机制有待进一步的阐明。真菌纤维素酶的分泌也是相当复杂的,在酶的分泌过程中,酶会发生例如糖基化等一系列变化。提高纤维素酶的活力测定方法的准确性,使其进一步规范化,是研究纤维素酶工作的另外一个重要的领域。随着现代生物学朝着分子水平的不断发展,分子生物学的许多方法也越来越多地被应用于纤维素酶的研究中,并取得了重要进展。对真菌纤维素酶合成调节机制的深入研究,将为提高纤维素酶产量,推动应用工作的发展打下理论基础。     

Simultaneous hydrolysis and fermentation of pulp mill primary clarifier sludge

Bioconversion of sludge from the primary clarifier of a sulphite pulping operation to ethanol offers a number of advantages over conventional disposal options. The amount of material which must be disposed of is reduced while, at the same time, salable and environmentally friendly fuel-ethanol is produced. In this study, primary clarifier sludge (PCS) was shown to be hydrolysed to produce fermentable sugars at a rate proportional to enzyme loading. Initial (1 hour) hydrolysis rates as high as 12.6 g reducing sugar/L · h were observed at an initial enzyme loading of 10 filter paper units (FPU)/g. Hydrolysis was inhibited by spent sulphite liquor (SSL), an inhibition which could be completely overcome by fermenting the SSL to remove sugars. Surfactants were found to only marginally improve the production of sugars. To reduce the deleterious effects of end product inhibition, single stage simultaneous hydrolysis and fermentation (SHF) was carried out using cellulase enzymes and Saccharomyces cerevisiae.     

纤维素酶活性测定的生物传感器研究

用TiO2凝胶包埋葡萄糖氧化酶电化学传感器(GOD/TiO2)测定纤维素酶(CE)的活性,即基于电极对纤维素水解后生成葡萄糖的催化氧化。考察了各种实验条件对CE酶活性测定的影响。实验表明,GOD/TiO2电极检测电位为+0.50Vvs.SCE,稳态响应电流与纤维素酶活性在10～300U/L之间有线性关系,该方法可用于微量羧甲基纤维素酶活性测定。     

纤维素酶的固体发酵

从来自国内外11株菌株的初步比较,筛选出酶活较高的康氏木霉N_2—78和瑞热埃木霉QM6a。对康氏木霉P_2和土曲霉3.3935所作的纤维素碳源试验中,发现木质纤维素较纯纤维素好,未经处理的木质纤维素比经处理的好。麸皮和稀土对康氏木霉P_2产酶作用影响较大,而对土曲霉3.3935则无明显的影响。     

黏胶纤维酶水解反应工艺研究

将黏胶纤维水解成糖类物质,是利用废弃黏胶纤维制备燃料乙醇的关键步骤.采用正交试验的方法,对纤维素酶催化黏胶纤维水解反应中的主要因素的影响进行了综合研究.结果表明,影响因素的显著性大小依次为:催化剂用量、固-液比、温度、pH值.对反应时间、温度、催化剂用量等影响因素进行的详细研究结果表明,酶的用量适当,反应温度选择在纤维素酶的适用温度上限,反应时间约为2 h是比较适宜的反应条件,黏胶纤维水解率约达87%.     

两种纤维素酶水解漂白针叶木纤维能力的比较

采用复合纤维素酶Celluclast 1.5L和内切纤维素酶Novozym 476等分别对漂白针叶木纤维进行了水解实验.通过对比两种酶的酶活力大小以及水解后纤维得率的变化情况,分析比较了两种纤维素酶水解纤维素纤维的能力大小.其结果显示:纤维素酶Celluclast 1.5L具有较高的滤纸酶活,而纤维素酶Novozym 476具有较高的CMC酶活.经纤维素酶Celluclast 1.5L水解处理后,随着酶用量的增加或酶处理时间的延长,纤维得率急速下降,当酶用量为20.0FPU/g、水解时间为2h时,纤维得率约为81%;当酶用量为10.0FPU/g、水解时间为48h时,纤维得率仅为55.34%,这说明纤维素酶Celluclast 1.5L对纤维素具有很强的水解作用,它不仅可以作用于纤维素的无定形区,同时也可以作用于纤维素的结晶区.纤维素酶Novozym 476亦能使纤维素发生一定程度的水解,但催化水解能力远低于纤维素酶Celluclast 1.5L,当Novozym 476用量为50.0CMCU/g时,仅有5%左右的纤维素水解成为葡萄糖.     

产纤维素酶菌株Pantoea ananatis P5的筛选鉴定和活性研究

对一种新型纤维素酶产生菌株P5进行了筛选鉴定和活性研究．该纤维素酶产生菌株在中国普通微生物保藏管理中心的保藏号是CGMCC NO．5356．利用产纤维素酶菌株筛选培养基,从土壤中筛选到一株产生纤维素酶菌株P5,对该菌株进行了分子鉴定和纤维素酶活性分析．结果显示,菌株P5分类上是Pantoea ananatis菌株,其纤维素酶适应温度和酸碱度范围较广,可应用于生物、食品、能源、纺织等领域．