Study on enzymatic hydrolysis of steam exploded straw by using shaking ball

A novel method of enzymatic hydrolysis was developed in this paper to produce a high conversion yield and hydrolysis rate. A comparison was described by using three methods of enzymatic hydrolysis and adsorption of steam exploded straw (SEWS): shaking ball in the regime, shaking with stirrer bed, and static state. The most adequate filter-paper activity, speed and reaction time were 3.6 × 10⁻²⁷ mol/(s·mL), 150 r/min and 24 h, respectively, with the reducing sugar yield of 0.43. The results showed that the method of shaking ball produced the highest adsorption, conversion yields and hydrolysis rate of the enzyme. This might be due to the continuous frequency increase of enzyme adsorption and desorption on the substrate surface as well as the relieved end-product inhibition. The morphological variation of SEWS was characterized by environmental scanning electron microscopy (ESEM). __________ Translated from Journal of Beijing University of Chemical Technology, 2006, 33(6): 26-30 [译自: 北京化工大学学报]     

NaOH-Fenton试剂预处理桑木的纤维素酶分段酶水解研究

采用分段酶水解木质纤维原料的方法,以NaOH-Fenton试剂预处理桑木为原料,通过在反应过程中及时移除葡萄糖和纤维二糖,减轻产物的抑制作用,最终达到提高酶水解得率和缩短酶解反应时间的目的。实验结果表明：纤维素酶用量为15FPIU/g（以纤维素计,下同）时,在三段（8+8+8h）水解过程中,经第一段水解,纤维素酶反应速率从1.25g/（L·h）提高到2.21g/（L·h）,第二段水解后,酶反应速率为1.54g/（L·h）,比未分段水解的酶反应速率提高了73%;当纤维素酶用量为40FPIU/g时,三段（8+8+8h）水解得率增至88.08%;三段（8+8+8h）水解充分利用了酶解残渣上的结合酶进行后续水解。对纤维素酶在预处理桑木上的吸附情况进行研究,发现桑木经NaOH-Fenton试剂预处理后,对纤维素酶的最大吸附量为8.08mg/g,预处理增加了纤维素酶与桑木间的吸附位点。     

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%。     

酶解反应与膜分离耦合连续制备酪蛋白磷酸肽

采用酶解反应与膜分离耦合新工艺连续水解全酪蛋白制备酪蛋白磷酸肽(CPPs)。考察了超滤膜对胰蛋白酶及底物溶液的截留效果;研究了初始底物质量浓度、初始酶质量浓度、反应体积、膜渗透通量等参数对反应器性能和反应转化率的影响规律;利用高效凝胶排阻色谱系统(HPSEC)对酶解产物进行检测分析;建立了酶膜反应器连续水解动力学模型,并对间歇与连续酶解过程进行比较分析,证明反应-分离耦合技术可使酶解效率及蛋白酶利用率大幅提高,并使产物得到调控与富集,为CPPs的酶法制备提供了一种更为有效的方法。     

NaOH/AQ预处理对玉米秸秆脱木质素及酶解性能的影响

对玉米秸秆进行氢氧化钠/蒽醌（NaOH/AQ）去木质化预处理,考察了预处理温度、时间和NaOH用量对玉米秸秆脱木质素程度的影响,并探讨了脱木质素程度对提高预处理后物料酶解性能的影响。L₉（3⁴）正交试验得出较适宜预处理工艺条件为：温度160℃,时间60 min,NaOH用量（以绝干原料质量计）2.8%;其他条件为AQ用量0.05%,固液比1：5（g：mL）,此时木质素脱除率为75%,酶解后聚糖转化率达到73.79%。随着物料脱木质素程度的提高,其酶解效率相应增加;当木质素脱除率达到一定程度后,预处理后的聚糖转化率达到最大值,继续提高木质素脱除率,聚糖转化率反而降低。响应面优化的酶水解工艺条件为纤维素酶用量30 FPU/g,β-葡萄糖苷酶10 IU/g,反应时间72 h,温度50℃,底物质量分数2.5%,此时还原糖得率为85.62%。对酶解液进行HPLC分析,酶解液中的葡萄糖质量浓度为14.83 g/L,木糖质量浓度为4.83 g/L。XRD分析显示,预处理前后纤维素的晶型没有变化,而结晶度由31.40%提高至46.91%,表明物料中木质素和半纤维素发生了不同程度的溶出。     

Use of membrane separation in enzymatic hydrolysis of waste paper

A three-stage process containing phosphoric acid pretreatment, enzymatic hydrolysis, and membrane filtration was performed on waste paper as a lignocellulosic material. In the first two stages, the effect of phosphoric acid concentration, enzyme loading, hydrolysis time, and substrate concentration on the amount of products was investigated. At the third stage using a proper membrane, the effect of substrate concentration and transmembrane pressure (TMP) on yield of the reducing sugars was studied. The novelty of the present study was to demonstrate the application of ultrafiltration membrane on the enzymatic hydrolysis process of waste paper. The reducing sugars concentration was determined by using the 3,5-dinitrosalicylic acid (DNS) reagent method. According to the results, a value of 0.5% was determined as the optimum concentration for phosphoric acid in the pretreatment stage. The reducing sugars yield was obtained as 67.4% in this concentration. Moreover, for the enzymatic hydrolysis of waste paper, the suitable amounts of cellulase enzyme loading and hydrolysis time were determined as 50 mg/g substrate and 48 h, respectively. In the filtration stage, increase of substrate concentration and decrease of TMP resulted in higher rejection of the reducing sugars. The experimental results revealed that the highest rejection was 19.2% at TMP of 3 bar and substrate concentration of 100 g/L.     

Optimization of the enzymatic hydrolysis conditions of steam‐exploded wheat straw for maximum glucose and xylose recovery

BACKGROUND: The enzymatic hydrolysis of steam‐exploded wheat straw using commercial enzyme complexes has been studied. A cellulase enzyme complex (Accellerase 1500), along with specific xylanase complements (Accellerase‐XC and Accellerase‐XY) provided by Genencor, have been used to enhance glucose and xylose recovery. A systematic study with response surface methodology (RSM) was used to check the effect of the operating conditions: pH (4–5), temperature (50–60 °C) and enzyme/substrate ratio (0.1–0.5 mL g_cellulose^?1) on the enzymatic hydrolysis with Acellerase 1500 to maximize the sugar yield. Xylanases were used as complements to increase the release of xylose. RESULT: Statistical results from ANOVA analysis demonstrated that the enzymatic hydrolysis was clearly improved by temperature and enzyme/substrate ratio. The optimum conditions for higher glucose and xylose releases were obtained with the higher enzyme dosage ratio (0.5 mL g^?1_cellulose), 50 °C and pH 4. CONCLUSION: Model validation at optimum operating conditions showed good agreement between the experimental results and the predicted responses for a confidence level of 95%. The use of the xylanase complements, Accellerase‐XY (accessory xylanase enzyme complex) and Accellerase‐XC (accessory xylanase/cellulase enzyme complex), increases the conversion of hemicellulose. Accellerase‐XC supplementation was more effective, obtaining an increase in yields of glucose and xylose of 11.8% and 23.6%, respectively, using a dosage of 0.125 mL g^?1_cellulose. © 2012 Society of Chemical Industry     

木质素对蔗渣酶水解的影响

采用机械活化方法对蔗渣进行处理,研究在原料可及性变化时木质素对蔗渣酶解的影响,用牛血清蛋白（BSA）预先使原料中木质素吸附饱和的方法来测定木质素对酶的吸附情况、用X-射线衍射和扫描电镜来测定蔗渣结晶结构、表面形态来表征原料可及性,从而分析影响的可能机制。结果表明木质素对蔗渣酶水解的影响与蔗渣可及性有关,原料可及性越高,影响越小。当蔗渣可及性相对较低时,此时木质素影响蔗渣酶水解转化率的两种方式（木质素与纤维素、半纤维素组成的致密结构限制纤维素对酶的可及性及木质素对纤维素酶的无效吸附作用）同时存在。当原料的可及性增加到一定程度后（如机械活化2h的蔗渣）,木质素对纤维素酶的吸附作用几乎消除,木质素对蔗渣酶水解的影响主要表现为对酶的可及性的限制。     

木质素结构以及表面活性剂对木质素吸附纤维素酶的影响

引言随着世界经济的快速发展和人口的增加,石油以及其他不可再生资源已经濒临枯竭。出于经济发展、国家安全以及环境保护等各个方面的需要,各个国家都在寻找石油的代替能源。生物乙醇这一新     

生物酶解法提取苦楝素工艺过程的研究

采用国产纤维素酶64001131,研究生物酶解法提取苦楝树皮中苦楝素的工艺条件。优化的工艺条件为:酶解液pH值4.5,酶解温度55℃,酶解时间1.5 h,酶浓度0.15 mg/mL。在此条件下,苦楝素的提取率达2.008%。     

外切-β-葡聚糖酶基因重组里氏木霉的筛选及其产酶性能

外切-b-葡聚糖酶是纤维素酶的重要组分之一,提高该组分的活力是增强纤维素酶协同降解性能、降低纤维素水解成本的关键。分别采用微晶纤维素琼脂平板法和滤纸崩解法,对已有的基因重组转化子进行筛选试验,获得了6个优良转化子,其滤纸崩解速率和微晶纤维素琼脂平板上的生长速率都较大。进一步在摇瓶条件下进行复筛试验,获得了外切-β-葡聚糖酶（C₁）高产转化子Trichoderma reesei ZU-101,液体培养48 h,其C₁酶活力可达18.24 U·ml^-1,是出发菌株的2.16倍;分析结果表明：重组转化子的纤维素酶体系中内切-b-葡聚糖酶和纤维二糖酶的活力与出发菌株相比变化不大,但由于外切-b-葡聚糖酶活力得到了大幅度提高,纤维素酶的总活力（滤纸酶活力FPA）也提高了61.9%。采用纤维素酶对碱预处理玉米秸秆进行酶解试验,当酶用量为20 FPIU·（g底物）^-1,水解48 h,重组转化子T.reesei ZU-101纤维素酶的酶解得率高达94.4%。本文的研究结果在可再生纤维素资源的生物转化与利用方面具有广阔的应用前景。     

Specific-enzyme release of cellulose-bound drugs,experimental and theoretical study

Various polymeric prodrugs of pholcodine were prepared by covalently linking a substrate ester of the hydroxylic drug to cellulosic derivatives. The ester derivatives of the drug were chosen so that the resulting insoluble cellulosic prodrugs became substrates of α-chymotrypsin. The hydrolysis of such compounds was studied for various spacer arms in conditions simulating the intestinal medium. A theoretical model was developed to describe the enzymatic catalysis of this hydrolysis, taking into account the adsorption of the enzyme onto the polymer, the hydrolysis reaction, and the enzyme denaturation. The resulting equations using independently determined parameter values such as the Langmuir adsorption constants, the enzyme denaturation constants, and the initial hydrolysis rate constant successfully correlated with the experimental data.     

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。     

绿液预处理对棉秆化学成分及酶水解糖化的影响

绿液的主要成分是Na2CO3和Na2S,可在硫酸盐浆厂碱回收系统中循环产生。将新疆棉秆在不同温度、硫化度和用碱量下经绿液预处理后,在不同酶用量下进行酶水解,并测定酶水解液中总糖得率以评价预处理效果。结果表明,原料预处理得率随用碱量的增加而下降。适当地提高蒸煮温度和用碱量,可以增加木质素脱除量,有利于后续酶水解的进行,但过于剧烈的预处理条件将导致碳水化合物的过度降解,因而降低酶水解糖的转化率。另外,由于新疆棉秆细小,棉秆皮占全秆比例较大,原料总糖含量较低,导致最终酶水解总糖得率偏低。在温度120?C,用碱量16%,硫化度32%条件下绿液预处理,酶水解糖转化率达到最大值,当酶水解酶用量为20 FPU/g(对浆料)时,葡聚糖、木聚糖和总糖的转化率分别为74.0%、61.3%和68.5%。     

Response surface methodology to optimize enzymatic preparation of deapio-platycodin d and platycodin d from radix platycodi

In the present work, we reported the enzymatic preparation of deapio-platycodin D (dPD) and platycodin D (PD) optimized by response surface methodology (RSM) from Radix Platycodi. During investigation of the hydrolysis of crude platycosides by various glycoside hydrolases, snailase showed a strong ability to transform deapio-platycoside E (dPE) and platycoside E (PE) into dPD and PD with 100% conversion. RSM was used to optimize the effects of the reaction temperature (35-45 °C), enzyme load (5-20%), and reaction time (4-24 h) on the conversion process. Validation of the RSM model was verified by the good agreement between the experimental and the predicted values of dPD and PD conversion yield. The optimum preparation conditions were as follows: temperature, 43 °C; enzyme load, 15%; reaction time, 22 h. The biotransformation pathways were dPE→dPD3→dPD and PE→PD3→PD, respectively. The determined method may be highly applicable for the enzymatic preparation of dPD and PD for medicinal purposes and also for commercial use.     

基于乳酸的深度共熔溶剂提取秸秆木质素对纤维素酶水解效率的影响

基于乳酸的深度共熔溶剂（DES）在脱木素和木质纤维素转化方面显示出巨大潜力。本文以乳酸为氢键供体（HBD）,氯化胆碱、盐酸胍、精氨酸、甜菜碱盐酸盐为氢键受体（HBA）合成了4种DES,并分别提取了木质素,对比研究了4种木质素对微晶纤维素酶水解效率的影响。结果表明,4种DES提取木质素的疏水性与其对酶蛋白吸附能力线性正相关,而两者与木质素与酶水解效率呈线性负相关性。非特异性吸附是该类木质素抑制纤维素酶水解效率最重要的因素,而疏水性作用则是引起酶蛋白吸附的主要原因。结构分析表明木质素的脂肪族羟基和酚羟基主导的氢键相互作用是促进酶蛋白吸附、降低酶水解效率的原因之一。此外,由羧基产生的静电斥力对纤维素酶水解效率的影响较弱。随着DES提取木质素能力的增强,木质素的羟基和羧基含量逐渐增高,其对纤维素酶的抑制作用增大。氯化胆碱-乳酸因其优良的木质素提取和生物质解构能力,提取的木质素对酶的抑制作用最强,其次是乳酸-盐酸胍和乳酸-甜菜碱盐酸盐,最后是乳酸-精氨酸。     

Enzymatic pretreatment on rose-hip oil extraction: Hydrolysis and pressing conditions

In a previous report [Zúñiga, M.E., J. Concha, C. Soto, and R. Chamy, Effect of the Rose Hip (Rosa aff. rubiginosa) Oil Extraction Cold-Pressed Process, in Proceedings of the World Conference and Exhibition on Oilseed Processing, and Utilization, edited by R.F. Wilson, AOCS Press, Champaign, 2001, pp. 210–213], the authors showed that an enzymatic pretreatment of rose-hip seeds, prior to oil extraction by cold pressing, improves the oil yield. In this work, we studied the effects of temperature and moisture during the enzymatic hydrolysis stage using two previously selected mixtures of commercial enzymes: (i) Olivex (mainly pectinase) plus Cellubrix (mainly cellulase), and (ii) Finizym (mainly β-glucanase) plus Cellubrix (mainly cellulase) (all from Novozymes A/S, Madrid, Spain). In addition, we evaluated the effect of enzymatic hydrolysis on the oil extraction pressing rate at different operational pressures. Samples hydrolyzed enzymatically by either of the two commercial enzyme mixtures at 45°C and 30–40% moisture showed oil extraction yields up to 60%, an increase of greater than 50%, as compared with control samples in which the enzyme solutions were replaced by water. Both the oil extraction rate and yield by pressing increased when enzymatic pretreatment was applied. The oil extraction yield increased slightly when the operation pressure was elevated; however, when the sample was preheated, the oil extraction yield was greatly increased, especially for enzyme-treated samples. Results confirmed the importance of temperature and moisture as enzymatic hydrolysis parameters that improve rose-hip oil extraction yields in the cold-pressing process. When pressing was carried out after preheating enzymatically treated samples, it was possible to increase the oil extraction yield to 72% compared with the control without preheating, which resulted in a 46% oil yield.     

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。     

Optimization of Enzymatic Hydrolysis of Chicken Fat in Emulsion by Response Surface Methodology

Chicken fat in an emulsion prepared with mechanical shearing and high pressure homogenization (HPH) was hydrolyzed using Candida cylindracea lipase. A homogenization pressure of 50 MPa, which can generate smaller droplets and higher hydrolysis efficiency than mechanical shearing, was fixed to prepare the emulsion for hydrolysis optimization. Response surface methodology (RSM) was applied to study the effect of temperature, enzyme loading, shaking rate and reaction time on the hydrolysis process. The results showed that all three-second-order polynomial models adequately fitted the experimental data. Additionally, hydrolysis parameters for the optimal yields of free oleic and linoleic acids were also obtained using the desirability function: a temperature of 38 °C, an enzyme loading of 0.48% (g/g fat basis), a shaking rate of 100 rpm and a reaction time of 80 min. Under the optimal conditions, the yields of free oleic and linoleic acids were predicted as being 0.470 and 0.118 g/g fat with recoveries of 94.6 and 93.7%, respectively. During the hydrolysis process, the particle size increased with concomitant boosting of the degree of hydrolysis and the stability of the emulsion system was gradually undermined by this reaction process.     

多元复合酶降解黄姜薯蓣皂苷元的工艺研究

研究了多元复合酶降解黄姜薯蓣皂苷工艺条件对黄姜薯蓣皂苷元收率的影响因素。结果表明：采用碱性果胶 酶+单宁酶+半纤维素酶+β-葡萄糖苷酶4元复合酶的组合形式,在酶用量为黄姜量的 1.5%,pH值为6.5,料液比为 1：4（g：g）,50℃ 下酶解 10 h,以乙醇为萃取剂的条件下进行的降解效果最好,薯蓣皂苷元的收率最高可达 4.34%。相比收率仅为 1.61% 的传统的酸解工艺,应用多元复合酶解工艺最高可使黄姜薯蓣皂苷元的收率提高1.69倍。