响应曲面法优化小麦秸秆纤维素酶水解条件

利用响应曲面试验设计方法(RSM),选择底物质量分数、酶投加量、温度、pH值及水解时间为试验因子,还原糖(RS)产率为响应值,考察小麦秸秆纤维素酶水解过程中各影响因子对还原糖产率的影响,对小麦秸秆纤维素酶水解条件进行优化。结果表明,所考察的5个影响因子对还原糖产率均具有显著影响(p<0.05)。所得回归方程R² 值为 0.946 9,p<0.05,变异系数(C_V)值为4.37%,足够精度值为26.396,说明模型高度显著,可以在设计范围内对响应值进行预测。模型预测最佳水解条件为底物质量分数8.0%,酶投加量为35 FPU/g(以秸杆质量计),温度50 ℃,pH值5.4,水解时间96 h。利用最优水解条件进行验证试验,所得还原糖产率为60.73%,水解液中葡萄糖和木糖质量浓度分别为31.84和 16.74 g/L。     

木质纤维素微波强化预处理方法的初步研究

以水稻秸秆为原料,采用微波辐射联合小分子有机酸预处理手段,考察了微波强度、酸质量分数、微波时间、固液比对木质素的移除和经预处理底物的酶水解效果的影响。通过正交实验确定了各因素的影响次序为:酸质量分数固液比微波时间微波强度。在最佳的预处理条件下(柠檬酸质量分数为20%,固液比为1∶20,微波时间为8 min,微波强度为600 W),酶水解96 h后的还原糖产率可达50.6%,比未处理的秸秆(22.5%)提高了28.1%。随后对最佳条件下处理的底物进行了一系列酶水解条件的优化,确定了酶水解的最优条件,即底物质量浓度为20 mg/m L,控制添加的酶的总蛋白量为20 mg,纤维素酶∶β-葡萄糖苷酶∶木聚糖酶蛋白量比为1∶1∶2,最佳表面活性剂吐温-80的质量分数为2%,此时的还原糖产率可达58.4%,比未优化的有明显的提高。     

离子及表面活性剂对甜高粱秆渣酶解的影响

为了提高纤维素酶水解经高温液态水处理后的甜高粱秆渣的效率,探讨了多种阴离子、阳离子以及吐温80(Tween 80)对纤维素酶活力的影响,并初步探讨了Tween 80影响甜高粱秆渣酶解的机制。酶激活试验表明,Br^-、I^-、NO₃^-、Ca²⁺、Mg²⁺和Co²⁺对纤维素酶有激活作用,但对甜高粱秆渣的水解效率提高不明显。添加Tween 80发现,随着浓度的增加,它对纤维素酶的抑制作用增强,而Tween 80添加量为0.175 ml·(g甜高粱秆渣)^-1时,甜高粱秆渣的酶解效率由16.6%提高到37.9%。吸附试验表明,甜高粱秆渣对纤维素酶和Tween 80的吸附达到一定限度后不再上升,Tween 80能显著降低甜高粱秆渣对纤维素酶的吸附。红外光谱分析发现,木质素对Tween 80的吸附要强于它对纤维素酶的吸附。     

复合酶法提取牡丹种皮黄酮类化合物研究

利用正交实验和单因素实验方法获取复合酶(纤维素酶、半纤维素酶、果胶酶)辅助乙醇提取牡丹种皮中黄酮类化合物的最佳复合酶比例和最佳酶解工艺条件;分析了不同的加酶方式对总黄酮类化合物提取量的影响。结果表明三种复合酶最优比例为(纤维素酶∶半纤维素酶∶果胶酶=4∶1∶3);最佳提取工艺条件为:温度47.5℃、p H=5、酶解1.5 h、液料比20∶1,总黄酮提取量可达64.78 mg/g;最优分步加酶比同时加酶提取量增加了4.58 mg/g,总黄酮提取量可达69.36 mg/g。     

纤维素酶高效水解麦秆的工艺研究

以纤维素酶水解蒸汽爆破麦秆的过程为研究对象,考察了底物浓度、纤维素酶用量、β-葡萄糖苷酶装载量以及化学激活剂对麦秆水解的影响。结果表明,高底物浓度下的最佳酶解工艺条件为底物(麦秆)浓度20%,酶装载量(U/g纤维素):滤纸酶活45、β-葡萄糖苷酶25、木聚糖酶800,0.1 mmol/L Mg2+、0.1 mmol/L Co2+、10 mmol/L Fe3+,1 g/L PEG2000、1 g/L Tween80和1 g/L山梨醇,搅拌速度120~150 r/min,分批补料,p H4.8,50℃,水解时间144 h。在此条件下,还原糖浓度达115.43 g/L,葡萄糖浓度达88.39 g/L,转化率也分别达到78.04%和88.73%。     

表面活性剂对木薯秆酶水解的影响

采用纤维素酶促水解的方法,以木薯秆为底物,研究了5种表面活性剂(SDS、Tween 20、Tween 80、Triton x-100、Triton x-114)及其用量对木薯秆酶水解得率的影响。结果表明,除SDS外,其他4种表面活性剂对木薯秆酶水解有不同程度的促进作用,Tween 20的效果最为显著。添加2.5 g/L的Tween 20水解72 h和5.0 g/L的Triton x-114水解60 h,酶水解得率分别达到了49.23%、49.48%,比空白样提高了33.45%和34.13%。     

固定化纤维素酶水解玉米秸秆的研究

以壳聚糖小球为载体固定纤维素酶,研究了固定化纤维素酶的酶学性质,并以经过蒸汽爆破预处理的玉米秸秆为底物,研究了固定化酶水解的最优条件.结果表明纤维索酶经过固定化后,其米氏常数为0.42 mg/mL,半衰期为162 h;酶解的最佳条件为固定化酶与底物的质量比为1:2.5、固液比为1:2.4(m/V)、温度为50℃,pH为5.0.秸秆水解率可达33%,纤维素水解率可达70%.     

1-十六烷基吡啶氯盐对纯纤维素酶解糖化的影响

以纯纤维素为底物,考察了阳离子表面活性剂1-十六烷基吡啶氯盐对纯纤维素酶解糖化的优化条件及其对体系酶活和酶动力学的影响。结果表明:当酶量为10 FPU/g,底物质量分数为1.25%,1-十六烷基吡啶氯盐质量浓度为5 g/L,反应时间为12 h时,纤维素平均转化率为59.91%,比不添加表面活性剂的空白样转化率提高了20.79%。1-十六烷基吡啶氯盐的加入,提高了酶解反应的最大反应速率和米氏常数,其中,最大反应速率提高了58.22%,米氏常数提高了101.42%。相对酶活提高率随时间呈现先升高后降低的规律,在24 h时,相对酶活提高率最高值达53.18%。     

一株产腈水合酶菌株酶活高效表达条件的研究

以抚顺腈纶厂废水中筛选的一株Nocardia sp为出发菌株,通过逐渐增加培养基中丙烯腈的体积浓度重复继代培养,得到1株酶活高效表达的Nocardia sp菌,酶活比出发菌株提高了15倍。确定了菌株培养的最优条件:葡萄糖(25 g/L),诱导剂脲(0.06 g/L)、Co2+(0.02 g/L)、丙烯酰胺(10%),反应条件pH(7)、温度(20℃),底物丙烯腈体积分数(<5%),产物丙烯酰胺的质量分数(<20%)。正交实验表明,反应温度和脲的加入量为反应过程中的显著因素。     

竹浆分批加酶工艺对长时间纤维素酶解性能的影响

优化纤维素酶水解工艺对于纤维素的高效利用具有重要意义。实验在保证纤维素酶相同的添加量时,采用多种方式分批添加纤维素酶水解竹浆,在长时间(7 d)的酶解过程中测定每天还原性糖产量,最后对长时间酶解过程中的影响因素进行了探究。实验结果表明:采用(0.35 mL+0.15 mL+0.1mL)添加工艺提高了酶解效率,酶解7d后糖产量提高了4.36%;纤维素酶随着培育时间的进行活性不断减小,培育1 d后的酶活性仅为原酶活性的53.87%;酶解糖产量随葡萄糖浓度的增加而降低,当葡萄糖浓度为30 mg/mL时,糖产量较无葡萄糖实验组降低了30.7%。酶解过程中酶活的丧失以及底物的抑制可能是影响纤维素酶长时间水解过程中的重要因素。     

Tween 80 enhancing cellulase recyclability during activation of dissolving pulp

Abstract

Enhancing cellulase recyclability is a promising approach to reducing the high enzyme cost for activation of dissolving pulp. For this purpose, Tween 80, an amphiphilic surfactant, was employed to facilitate cellulase recovery from fibers. Results indicated that the Cellulase–Tween complex was formed (with a particle size increase of 140%) through a hydrophobic interaction. A much lower maximum binding capacity and affinity (i.e., absorption ability) of cellulase onto fibers (6.03 versus 24.3?mg protein/g cellulose and 7.45 versus 13.8?ml/mg protein) were achieved for the complex in comparison with the control. The recovered cellulase activity was increased up to 65.7% from the control one in five consecutive recycle rounds with Tween 80 addition. As a result, the recovered cellulase efficiency was improved in terms of viscosity and reactivity. Other properties, such as α-cellulose content, alkaline solubility, molecular weight distribution, morphology, and crystalline structure, were all supported by the positive effect of Tween 80 in cellulase recycling.     

Effects of natural oils and surfactants on cellulase production and activity

Emulsification of natural oils by a surfactant increases their efficiency as chemical antifoams. The presence of low concentrations of fatty acids or other surfactants have been reported to inhibit or stimulate microbial growth and/or product formation. The effects of different natural oils, Tween 80 and saponin on growth and cellulase production by T. reesei and S. pulverulentum have been investigated. It was found that in general, emulsification leads to higher cellulase activities in both cultures, though there are variations in enzyme levels depending on the presence or absence of Tween 80 and of different oils in growth media as well as the substrates used for cellulase assay.     

Tween80对稻草水解及同步糖化与发酵产乳酸的影响

在生物转化纤维原料产乳酸的过程中,酶解纤维原料产还原糖是限速步骤。为了获得较高的产物产率,需较高的酶用量,这使大规模酶解废弃纤维原料的成本很高。对吐温80在酶解稻草纤维素产糖,以及耐高温乳酸菌同步糖化发酵稻草产乳酸过程中的作用进行了考察。初步结果表明,吐温80加入可使保持同等程度的水解率所需的酶用量降低,添加0.2 g/g底物的吐温80到酶用量10 FPU/g体系,水解120 h的糖产率为292.2 mg/g,比不加表面活性剂体系的糖产率增加了11%;添加0.7 g/L的吐温80进行同步糖化与发酵72 h,能使乳酸产量提高24.2%。     

麦秸秆的氢氧化钙预处理及酶解试验研究

采用氢氧化钙对麦秸秆进行预处理,以酶解还原糖得率为目的,分别优化预处理及酶解条件。结果表明,氢氧化钙预处理麦秸秆的最佳条件是:Ca(OH)2添加量为0.06g/g(对秸秆),固液比为1:10,在120℃下反应时间为2h;最佳酶解条件是:温度50℃,pH4.8,纤维素酶17FPU/g(对秸秆),木聚糖酶160IU/g,在添加0.15g/g(对秸秆)Tween80条件下,酶解液中还原糖质量浓度为62.32g/L,酶解还原糖得率达85.23%。     

Effect of surfactants on α‐amylase production in a solid substrate fermentation process

Solid substrate fermentation (SSF) is a process where the substrate is a moist solid, which is insoluble in water but not suspended in water. In this study SSF of Bacillus subtilis (ATCC 21556) was used to produce an enzyme of commercial importance, α‐amylase, using as a substrate potato peel. To enhance the production of this enzyme, two nonionic synthetic surfactants were used, Tween 80 and Tween 20, one anionic surfactant, SDS at concentrations of 0.05% and 0.10% (v/w) and a biosurfactant produced by Bacillus subtilis (ATCC 21332), known as surfactin, at concentrations of 0.003%, 0.007%, 0.013% and 0.03% (w/w). The results have shown that surfactants significantly increase the production of α‐amylase. Tween 80 at 0.10% and surfactin at 0.013% provided the highest enzyme activity when compared with the control. © 1999 Society of Chemical Industry     

反相乳液聚合法合成阳离子聚丙烯酰胺的研究

以液体石蜡为连续相，选择Span80-Tween80的复合乳化剂，进行了AM—DMC的反相共聚合反应，研究了温度、单体配比、乳化剂质量分数对聚合物性能的影响。     

白油W/O/W型多重乳状液的稳定性研究

以多重乳状液相对体积为衡量标准,用显微镜直接观察,探讨了乳化剂的HLB值、质量分数、亲油亲水乳化剂体积比及油水的相比等对白油W/O/W型多重乳状液体系稳定性的影响。结果表明单一乳化剂体系中适宜的制备条件:乳液中乳化剂质量分数为12.2%,V(Span80)/V(Tween80)=7.5;适合多重乳液稳定的油水相比为:第一相体积比为2.5,第二相体积比为0.2。复合乳化剂体系中适宜的制备条件:第一相乳化剂的HLB值为6.5,V(复合乳化剂)/V(Tween80)=27.5,乳液中乳化剂质量分数为9.5%。     

An approach to optimization of enzymatic hydrolysis from sugarcane bagasse based on organosolv pretreatment

BACKGROUND: The organosolv pretreatment followed by enzymatic hydrolysis of the pretreated material and subsequent fermentation of the hydrolysate produced, was the strategy used for ethanol production from sugarcane bagasse. The effect of different operational variables affecting the pretreatment (the catalyst type and its concentration, and the pretreatment time) and enzymatic hydrolysis stage (substrate concentration, cellulase loading, addition of xylanase and Tween 20, and the cellulase/β‐glucosidase ratio), were investigated. RESULTS: The best values of glucose concentration (28.8 g L^?1) and yield (25.1 g per 100 g dry matter) were obtained when the material was pretreated with 1.25% (w/w) H₂SO₄ for 60 min, and subsequently hydrolyzed using 10% (w/v) substrate concentration in a reaction medium supplemented with xylanase (300 UI g^?1) and Tween 20 (2.5% w/w). Fermentation of the broth obtained under these optimum conditions by Saccharomyces cerevisiae resulted in an ethanol yield of 92.8% based on the theoretical yield, after 24 h. CONCLUSION: Organosolv pretreatment of sugarcane bagasse under soft conditions, and subsequent enzymatic hydrolysis of the pretreated material with a cellulolytic system supplemented with xylanase and Tween 20, is a suitable procedure to obtain a glucose rich hydrolysate efficiently fermentable to ethanol by Sacharomyces cerevisiae yeasts. Copyright © 2010 Society of Chemical Industry     

DMDAAC-AM-MA反相微乳液聚合体系的制备与优化

以二甲基二烯丙基氯化铵(DMDAAC)、丙烯酰胺(AM)和顺丁烯二酸(MA)为乳化单体,正丁醇为助乳化剂,聚氧乙烯辛基苯酚醚-10(Triton X-10)、失水山梨醇油酸酯(Span 80)和聚氧乙烯山梨醇酐硬脂酸酯(Tween 60)为乳化剂,液体石蜡、异辛烷和环己烷为有机溶剂,进行体系增溶实验。采用电导法和染料法研究了单体总质量浓度、AM摩尔分数、阴阳离子单体配比、乳化剂质量分数、助乳化剂与复合乳化剂质量比等对微乳液体系最小乳化剂用量(Smin)和最大增溶水量的影响。结果表明,异辛烷与Span80/Tween60复合乳化剂组成的乳化体系增溶性较好;当单体总质量浓度为40%、n(AM)∶n(MA)∶n(DMDAAC)=6∶2∶2、亲水亲油平衡值(HLB值)为11.30、w(复合乳化剂)=33%和m(助乳化剂)∶m(复合乳化剂)=1∶7时,所形成的反相微乳液增溶水量大、乳化剂用量少且稳定性强。