Studies on preparation technology of nanometer pearl powder by enzyme digestion

以珍珠粉粒径为考察指标,采用食品工业用酶处理,研究了蛋白酶种类、酶解时间、加酶量与珍珠粉粒径分布范围的关系.实验表明:10 g珍珠粉,加入50 mL去离子水溶解,使用饱和Ca( OH)2溶液调pH值至11.0,再加入0.3g碱性蛋白酶,于40℃水浴保温酶解6h.酶解后约有60％珍珠粉粒径分布小于1μm,已满足国家标准(征求意见稿)的规定要求,为纳米级珍珠粉.     

脱脂沙棘籽制备活性多肽的研究北大核心CSCD

本文比较了用巯基蛋白酶酶解沙棘籽蛋白所得酶解多肽对乙醇脱氢酶（ADH）的激活率效果。结果表明,木瓜蛋白酶的酶解产物对ADH的激活效果最好。该活性多肽的最佳制备条件为,酶解pH为6．5,温度为50℃,酶解时间3．5h,加酶量为4000U／g。体外实验表明,木瓜蛋白酶水解产物中ADH激活率较高的为500—2000Da的多肽,达57．52％,与未分级的酶解液相比,活性提高显著（P〈0．01）。分子量在2000～3000Da的沙棘多肽也具有一定的ADH激活率,为35．09％,说明具有ADH激活率的活性多肽分子量为500～3000Da。木瓜蛋白酶酶解液经膜分离处理后,收集得到的500—3000Da的多肽组成占总肽及氨基酸质量的66．50％。     

响应面法优化蚕豆蛋白酶解工艺条件的研究

考察了碱性蛋白酶、胰蛋白酶和中性蛋白酶对蚕豆蛋白的酶解效果,探讨了水解度（DH）与酶解产物抗氧化活性间的关系。通过单因素试验和响应面分析法,得到碱性蛋白酶酶解工艺的最佳条件。结果表明,温度50℃、pH8.0、酶底比8%、底物浓度3%条件下酶解3h,水解度0~22%内,碱性蛋白酶较胰蛋白酶和中性蛋白酶水解蚕豆蛋白效果好;DH与还原能力（R2=0.68~0.81）及ABTS清除能力（R2=0.98~0.99）具有较好的相关性,碱性蛋白酶酶解液较其他2个酶解液有较好的还原能力和ABTS清除能力;优化后的最佳酶解工艺参数为：酶底比8%,温度50℃、pH 7.6,对蚕豆蛋白还原能力的影响顺序为酶底比＞pH＞温度;在此条件下,蚕豆蛋白酶解液的还原能力理论值为0.174,验证试验测得还原能力为0.173,与理论值接近。     

利用N糖苷酶F对单克隆抗体N糖酶解条件的优化

为了优化利用N糖苷酶F（PNGase F）酶解单克隆抗体中N糖的方法，应用本公司生产的单抗对PNGase F酶的酶解条件进行优化，包括缓冲液pH、酶种类、仪器、酶解程度、变性缓冲液及酶加入量等，总结酶解条件对N糖谱结果的影响。结果显示，置换缓冲液至1×PBS中可以避免某些单抗在低pH酶解时G0F转化为G0F-GN并可改善峰型；快速PNGase F和加入变性缓冲液能有效提高酶解效率；UPLC和1.7 μm粒径色谱柱能提高分离度；不完全酶解影响N糖含量结果。研究结果表明，采用优化的酶解条件可快速、有效的酶解单抗上的N糖，使N糖谱结果准确可靠，为细胞株筛选和单抗药物质量控制提供有效手段。     

耐有机溶剂蛋白酶产生菌Bacillus cereusWQ9—2发酵条件及酶学性质

以蜡状芽孢杆菌Bacillus cereus WQ9—2为产耐有机溶剂蛋白酶的出发菌株,对其产酶条件进行优化并初步研究了其酶学性质。在单因素实验基础上,通过中心复合实验确定了产酶的最佳发酵条件为酵母粉8g/L,葡萄糖17g/L,KH2P040．5g／L,无水MgS040．3g／L,CaCl20．5g／L,NaCl1．0g/L;pH7．5。实验中发现采用低温发酵能大大缩短菌体产酶周期;通过优化发酵时间由最初84h缩短到48h,最高比酶活为3921U／mL。金属离子螯合剂1,10菲罗啉（1,10-phenanthroline）、乙二胺四乙酸（EDTA）对该酶有较强的抑制作用,表明该酶可能为金属蛋白酶;Ca2＋对该酶的活力及热稳定性有显著提高作用。     

响应面法优化黄绿蜜环菌子实体蛋白酶解条件

黄绿蜜环菌（Armillaria luteo—virens Sacc）子实体中蛋白含量很高,采用酶水解黄绿蜜环菌子实体蛋白的方法是生产具有生物学功能特性的活性肽的有效途径。实验采用酸性蛋白酶、中性蛋白酶和碱性蛋白酶水解黄绿蜜环菌子实体蛋白制备生物活性肽。实验中确定了酶解前处理条件,即最适宜的酶制剂为中性蛋白酶。酶解条件的优化采用中心组合响应面分析法：建立数学模型回归分析,模型评价,最后进行验证实验。结果表明：底物浓度为7．6％、加酶量为1．6％、酶解温度为52％、酶解时间为6．4h,此条件下蛋白水解度最高。     

高效液相色谱法在脱氧核糖核酸酶解动力学研究中的应用

为了更好地对脱氧核糖核酸酶解动力学过程进行研究,建立脱氧核糖核酸（DNA）酶解液中4种脱氧核苷酸（腺嘌呤脱氧核苷酸（dAMP）、鸟嘌呤脱氧核苷酸（dGMP）、胞嘧啶脱氧核苷酸（dCMP）、胸腺嘧啶脱氧核苷酸（dTMP））的高效液相测定方法,能将酶解液中4种脱氧核苷酸完全分离并准确定量。在此基础上,对DNA酶解的动力学进行初步研究,其反应机理为不存在底物和产物抑制的双底物顺序反应,动力学方程为x=1／bin（1＋abt）（其中a=0．3723p0—0．974;b=-0．0493p0^2＋1．1150p0-1．1103）,该方程可以很好地描述DNA酶解过程,误差仅为3．31％.     

内生青霉菌纤维素酶辅助提取槐米总黄酮

研究内生青霉菌（Penicillium sp．B-4）胞外纤维素酶在槐米总黄酮提取中的辅助应用。内生青霉菌在起始pH4,5的综合马铃薯培养基中,150r／min,40℃下摇瓶,培养7d,具有较高的纤维素酶比活力（3．57U／mL）。槐米干粉投入青霉菌发酵液中进行酶解处理,比较酶料比、酶解温度、酶解时间和酶解液pH对槐米总黄酮提取率的影响,发现槐米干粉以酶料比40：1（mL／g）加入粗酶液中,在pH4．5、温度40℃下酶解处理1h后,黄酮提取率可达12．2％,比常规提取率增加了38．7％。内生菌纤维素酶辅助提取法为槐米黄酮提取的可行新方法。     

碱性蛋白酶产生菌的筛选及发酵条件考查

从土壤中分离到的167株芽胞杆菌中选出一株高产、稳产碱性蛋白酶菌株A4－3（嗜碱性枯草芽胞杆菌）。该菌株最适产酶条件为（g／100ml）：蔗糖6．0;豆饼水解液10.0;Na2CO31．0;起始pH9．5。在该条件下,经37℃振荡培养48h,酶活力达2612u／ml。     

海藻酸钠包埋法制备固定化菠萝蛋白酶

以海藻酸钠为载体,包埋法固定菠萝蛋白酶,对固定化奈件进行优化,同时探讨固定化菠萝蛋白酶的部分酶学性能。结果表明：固定化菠萝蛋白酶的质量受海藻酸钠质量分数、固定化酶量、固定化时间以及CaCl2质量分数的影响,其最佳固定化条件为：海藻酸钠质量分数1．0％,CaCl2质量分数3％,固定化酶液量与海藻酸钠体积之比1：2,固定化时间60min,在此条件下,制备的固定化菠萝蛋白酶的比活力为211．8U／g（湿质量载体）,由此制得的固定化酶的最适pH为7．6,与游离酶相比,升高了0．8个pH单位,同时显示固定化菠萝蛋白酶能耐受较高的碱性环境,固定化酶最适温度与游离酶相同,均为50℃,固定化酶在较高温度范围内,仍能保持较高的相对活力。     

Comparison of the rates of enzymatic hydrolysis of pretreated rice straw and bagasse with celluloses

The rates of enzymatic hydrolysis of pretreated rice straw and bagasse have been studied and compared with the hydrolysis rates of microcrystalline cellulose powder (MCCP) and Solka Floc. The effects of particle size reduction and enzyme loading on the rates of hydrolysis of rice straw and bagasse were also studied. It was found that the rates of hydrolysis of pretreated rice straw and bagasse are much higher than that of MCCP and Solka Floc. For both rice straw and bagasse, particle size reduction had very little effect in enhancing the rate of hydrolysis. Lignin present at <10% did not seem to hinder the accessibility of the enzyme to the cellulose surface. An enzyme loading > 40 Ug^?1 had no effect on the hydrolysis rate of rice straw or bagasse.     

不同方法提取杜仲中桃叶珊瑚苷等4种高活性成分的比较研究

为研究生物化学方法对杜仲中高活性成分的提取率,本文以高效液相色谱法检验样品中提取出活性成分的含量差异,比较了在相同条件下酶解法、水提法、醇提法对杜仲树皮中桃叶珊瑚苷、京尼平苷、京尼平苷酸与绿原酸这4种活性成分的提取量,并对4种活性成分的不同提取效果进行了比较研究。结果发现三种方法的提取量存在明显差异,其中桃叶珊瑚苷提取量为:酶解法22. 42μg/g,水提法8. 27μg/g,醇提法9. 13μg/g;京尼平苷提取量为:酶解法77. 89μg/g,水提法33. 19μg/g,醇提法7. 76μg/g;京尼平苷酸提取量为:酶解法110. 05μg/g,水提法36. 63μg/g,醇提法47. 40μg/g;绿原酸提取量为:酶解法345. 35μg/g,水提法118. 85μg/g,醇提法172. 04μg/g,即酶解法提取量均比水提法、醇提法高出数倍。实验表明酶解法是3种方法中提取效果最好的,且无化学污染。     

Effect of particle size on the rate of enzymatic hydrolysis of cellulose

The effect of particle size on enzymatic hydrolysis of cellulose has been investigated. The average size of microcrystalline cotton cellulose has been reduced to submicron scale by using a media mill. The milled products were further subjected to hydrolysis using cellulase. High cellulose concentration (7%) appeared to retard the size reduction and resulted in greater particles and smaller specific surface areas than those at low concentration (3%) with the same milling time. Initial rate method was employed to explore the rate of enzymatic hydrolysis of cellulose. The production rate of cellobiose was increased at least 5-folds due to the size reduction. The yield of glucose was also significantly increased depending upon the ratio of enzyme to substrate. A high glucose yield (60%) was obtained in 10-h hydrolysis when the average particle size was in submicron scale.     

Efficient Acetone–Butanol–Ethanol Production from Corncob with a New Pretreatment Technology—Wet Disk Milling

Acetone–butanol–ethanol (ABE) production from corncob was achieved using an integrated process combining wet disk milling (WDM) pretreatment with enzymatic hydrolysis and fermentation by Clostridium acetobutylicum SE-1. Sugar yields of 71.3 % for glucose and 39.1 % for xylose from pretreated corncob were observed after enzymatic hydrolysis. The relationship between sugar yields and particle size of the pretreated corncob was investigated, suggesting a smaller particle size benefits enzymatic hydrolysis with the WDM pretreatment approach. Analysis of the correlation between parameters representing particle size and efficiency of enzymatic hydrolysis predicted that frequency 90 % is the best parameter representing particle size for the indication of the readiness of the material for enzymatic hydrolysis. ABE production from corncob was carried out with both separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes using C. acetobutylicum SE-1. Interestingly, when considering the time for fermentation as the time for ABE production, a comparable rate of sugar consumption and ABE production in the SHF process (0.55 g/l·h sugar consumption and 0.20 g/l·h ABE production) could be observed when glucose (0.50 g/l·h sugar consumption and 0.17 g/l·h ABE production) or a mixture of glucose and xylose (0.68 g/l·h sugar consumption and 0.22 g/l·h ABE production) mimicking the corncob hydrolysate was used as the substrate for fermentation. This result suggested that the WDM is a suitable pretreatment method for ABE production from corncob owing to the mild conditions. A higher ABE production rate could be observed with the SSF process (0.15 g/l·h) comparing with SHF process (0.12 g/l·h) when combining the time for saccharification and fermentation and consider it as the time for ABE production. This is possibly a result of low sustained sugar level during fermentation. These investigations lead to the suggestion that this new WDM pretreatment method has the potentials to be exploited for efficient ABE production from corncob.     

Effect of surfactant and particle size reduction on hydrolysis of deinking sludge and nonrecyclable newsprint

Disposal of sludge from deinking mills represents a significant proportion of operating costs. Bioconversion of the cellulosic fraction of deinking sludge (DIS) to ethanol greatly reduces disposal costs while producing an environmentally friendly fuel. In this study, the cellulosic fraction of newsprint and deinking sludge was hydrolysed to produce fermentable sugars. For newsprint, a particle size of 1 to 1.5 mm provided optimal reaction rates in batch reactors over practical hydrolysis times, and reducing sugar concentrations as high as 35 g/L could be achieved using a fed-batch reactor configuration. For both newsprint and DIS, the hydrolysis rate increased nonlinearly with enzyme loading. Tween-80 only marginally improved sugar production but was able to release sugars from cellulosic substrates in the absence of lytic enzymes, in an amount proportional to the surfactant concentration and the substrate particle size. DIS was relatively recalcitrant to enzymatic hydrolysis, possibly due in part to inhibition by hydrophobic constituents. (c) 1995 John Wiley & Sons, Inc.     

Particle size distribution of rice flour affecting the starch enzymatic hydrolysis and hydration properties

Rice flour is becoming very attractive as raw material, but there is lack of information about the influence of particle size on its functional properties and starch digestibility. This study evaluates the degree of dependence of the rice flour functional properties, mainly derived from starch behavior, with the particle size distribution. Hydration properties of flours and gels and starch enzymatic hydrolysis of individual fractions were assessed. Particle size heterogeneity on rice flour significantly affected functional properties and starch features, at room temperature and also after gelatinization; and the extent of that effect was grain type dependent. Particle size heterogeneity on rice flour induces different pattern in starch enzymatic hydrolysis, with the long grain having slower hydrolysis as indicated the rate constant (k). No correlation between starch digestibility and hydration properties or the protein content was observed. It seems that in intact granules interactions with other grain components must be taken into account. Overall, particle size fractionation of rice flour might be advisable for selecting specific physico-chemical properties.     

Particle morphology characterization and manipulation in biomass slurries and the effect on rheological properties and enzymatic conversion

An improved understanding of how particle size distribution relates to enzymatic hydrolysis performance and rheological properties could enable enhanced biochemical conversion of lignocellulosic feedstocks. Particle size distribution can change as a result of either physical or chemical manipulation of a biomass sample. In this study, we employed image processing techniques to measure slurry particle size distribution and validated the results by showing that they are comparable to those from laser diffraction and sieving. Particle size and chemical changes of biomass slurries were manipulated independently and the resulting yield stress and enzymatic digestibility of slurries with different size distributions were measured. Interestingly, reducing particle size by mechanical means from about 1 mm to 100 μm did not reduce the yield stress of the slurries over a broad range of concentrations or increase the digestibility of the biomass over the range of size reduction studied here. This is in stark contrast to the increase in digestibility and decrease in yield stress when particle size is reduced by dilute-acid pretreatment over similar size ranges.     

不同酶对草鱼蛋白功能特性及风味成分的影响

通过蛋白酶对草鱼进行酶解,对其酶解产物的功能特性和酶解液的风味成分进行研究。结果显示,风味蛋白酶、胰蛋白酶酶解产物的溶解性、热稳定性均优于胃蛋白酶酶解产物(P0.05)。经酶处理后鱼肉酶解液鲜甜味氨基酸占比增加,苦味氨基酸占比减少;利用SPME-GC-MS共检测出83种挥发性成分,以醛酮类和醇类为主,酶解处理后醇类的相对含量显著减少,醛酮类的相对含量则显著增加,酶解液具有独特风味。经胰蛋白酶、风味蛋白酶酶解后的草鱼肉酶解产物及酶解液均可作为食品基料应用于加工中。     

鲜鹿茸酶降解过程的研究

研究了胰蛋白酶、Alcalase 碱性蛋白酶、木瓜蛋白酶对鲜鹿茸的降解过程,确定了优化降解工艺条件,具有一定的理论意义和实践价值。确定了Alcalase 碱性蛋白酶的降解效率最高,通过单因素实验确定了降解过程中底物浓度、酶解温度、pH值和酶解时间为影响鲜鹿茸降解率的主要因素。正交试验确定最佳的酶解条件为:底物浓度0.08 g/ml、酶解温度65 ℃、pH 9.0、酶解时间6.0 h。在此条件下,鲜鹿茸降解率高达92.6%,氨基酸产品收率达12.1%。     

Micronization of lysozyme by supercritical assisted atomization

Supercritical Assisted Atomization (SAA) has been used to produce lysozyme microparticles. Lysozyme has been micronized using water, buffered water at pH 6.2 and water–ethanol mixtures at different volume percentages. Precipitated lysozyme particles were spherical, with a narrow particle size distribution (PSD) ranging from 0.1 to 4 µm. The concentration of lysozyme in the liquid solvent mixture had a nonlinear effect on the particle distribution, with an increase of the X_0.9 from about 1 to 3 µm varying the enzyme concentration from 5 to 20 mg/mL. Precipitation temperature was set as low as possible to avoid enzyme degradation. High‐performance liquid chromatography analysis showed no degradation of lysozyme and the enzyme activity, measured by turbidimetric enzymatic assay, only slightly decreased after SAA processing. Depending on the process conditions lysozyme retained from 95% to 100% of the biological activity compared to the untreated enzyme. Biotechnol. Bioeng. 2009; 104: 1162–1170. © 2009 Wiley Periodicals, Inc.