醋酸棉酚对土壤脲酶动力学特性的影响

通过不同浓度醋酸棉酚存在下的土壤脲酶催化尿素水解试验,研究醋酸棉酚对土壤脲酶的抑制作用,得到酶促反应的降解动力学参数和有关的热力学参数.结果表明,醋酸棉酚对土壤脲酶活性的抑制能力随醋酸棉酚浓度的增大而增强.由尿素浓度的倒数和水解速率的倒数为横纵标的双倒数曲线得到米氏常数(Km)和最大速率(Vmax),随着醋酸棉酚浓度的增大,酶促反应的Km和Vmax降低.在温度30℃时,Km由3.918 mol/L降到1.164 mol/L,Vmax由1.185×10-4 mol/(g·h)降到5.6×10-6 mol/(g·h).在醋酸棉酚的影响下,酶促反应的热力学参数包括活化能(Ea)、活化焓变(△H)和温度系数(Q10)有所下降.     

适宜无机盐及浓度抑制麦胚脂肪酶存在时油水界面张力

为了探明无机盐稳定化脂肪酶的潜在机理,该文以小麦胚芽脂肪酶为研究对象,基于界面酶学的分析方法,研究添加浓度介于1.0×10-9~1.0×10-2 mol/L的Na~+、K+、Ca~(2+)、Mg~(2+)的氯化物对小麦胚芽脂肪酶存在的油-水界面特性的影响。结果表明,当小麦胚芽脂肪酶体系浓度为1.7×10-6 mol/L时,一价金属中Na~+更有利于抑制油-水界面的表面张力(P0.05),对麦胚脂肪酶存在时的油水界面特性影响也较大;二价金属离子Ca~(2+)对界面张力的影响趋势与一价离子不同,在高浓度时反而增加界面张力;当油水界面上存在脂肪酶的催化底物(三油酸甘油酯)和产物(油酸)时,添加浓度分别为10-6、10-6、10-4和10-9mol/L的Na~+、K~+、Ca~(2+)、Mg~(2+)均可一定程度上降低油水界面张力,从而降低麦胚脂肪酶的作用效果,三油酸甘油酯存在时Mg~(2+)的作用效果最明显(P0.05),油酸存在时Na+的作用效果最明显(P0.05)。综上,无机盐金属离子主要通过影响麦胚脂肪酶在油水界面的聚集行为及底物结构状态起到钝化麦胚脂肪酶的作用,该结果可为麦胚脂肪酶存在时界面的活性调控及麦胚的稳定化处理提供参考。     

暗棕壤吸附铜离子特征及其影响因素研究

应用平衡吸附法,研究了不同pH、离子强度、温度、Cu~(2 )浓度和接触时间条件下,暗棕壤对Cu~(2 )的吸附作用,并利用热力学和动力学方程对实验结果进行了拟合。结果表明:(1)随pH提高,Cu~(2 )的吸附率增加,pH 8时的吸附率约为91%。(2)随体系离子强度增加,Cu~(2 )的吸附量先下降后增加。(3)随溶液中Cu~(2 )浓度增加,Cu~(2 )在暗棕壤上的吸附量也增加,并且在低浓度时增加比较迅速。吸附等温线可用Frendlich,Langmiur和Temkin方程很好的描述,其中又以Langmiur方程拟合程度最好。根据Langmiur方程中平衡常数(k_L)得到的热力学参数指出,反应是自发的、吸热的,但增温却没有改变Cu~(2 )在暗棕壤表面的混乱度。(4)随接触时间延长,暗棕壤对Cu~(2 )的吸附量也增大,具体表现为吸附开始的快速反应阶段和经过一段时间后的慢速反应阶段。双常数能更好地拟合暗棕壤对Cu~(2 )的吸附动力学曲线,根据一级动力学方程的反应速率常数(k_D),计算出了Cu~(2 )吸附的活化能和活化热力学参数,指出温度升高有利于提高反应的速率。     

酶法有限水解含油菜籽蛋白的机理及动力学

为探索油菜籽水相酶解法萃取菜籽油的机理,以Alcalase蛋白酶在温度50℃、pH8.0条件下,对以水剂法从菜籽中提取的含油菜籽蛋白进行酶解处理.试验研究结果表明:含油菜籽蛋白乳液的固形物颗粒度对酶解反应速率有明显影响,总的趋势是颗粒度越小,酶促反应速率越大;酶催化水解速率随水解进程呈指数下降,在反应过程中过高的底物浓度会抑制酶的失活.在此基础上由试验数据推导出描述催化水解含油菜籽蛋白的动力学方程,由此通过控制酶与底物浓度之比、反应时间,可以控制水解作用的程度,从而指导和优化菜籽水相酶解法提取菜籽油的工艺.     

双温度计法测定几种矿物与硼反应的热效应

量热法对吸附热的测定可直接了解多种反应的特性变化情况。用专门的微量热计已测定了某些摩尔反应焓[1] ,土壤胶体在吸附重金属离子时的能量变化[2 ] ,生化反应的米氏常数,反应速率常数,并可对酶 底物过渡态结构进行论证[1] 。用热电偶方法可测定钠蒙脱,钙蒙脱的悬浮液对尿素、     

纳米TiO2对土壤Hg2+吸附解吸性能的影响

为了研究纳米TiO_2对土壤吸附解吸汞性能的影响,以三峡水库消落区土壤为代表,选择2种晶型(锐钛矿和金红石)的纳米TiO_2颗粒,设置3个浓度梯度(2,4,8g/kg),制成含有不同浓度纳米TiO_2颗粒的土样,配制不同浓度的Hg~(2+)溶液,进行吸附解吸试验。结果表明:纳米TiO_2颗粒可提高土壤对Hg~(2+)的吸附量,但受颗粒的晶型和浓度的影响;锐钛矿颗粒对土壤吸附解吸Hg~(2+)的影响更大,其吸附作用力更强,不易解吸,其中4g/kg处理组吸附作用最强,与对照相比,最大吸附量可提高32.65%,且其解吸率低于对照组;金红石颗粒处理组对Hg~(2+)的吸附量随颗粒浓度的升高而增大,但吸附作用力较弱,容易被解吸,8g/kg处理组影响最大,与对照相比,最大吸附量提高18.18%,但其解吸率也最大。因此,纳米TiO_2颗粒增强土壤对汞的吸附作用,从而可能影响汞在环境中的迁移转化过程,特别是锐钛矿型颗粒。     

肌原纤维蛋白浓度对风味物质吸附能力的影响

为探究肌原纤维蛋白浓度对风味物质的作用机制,本研究建立了肌原纤维蛋白和风味物质作用的复合体系,研究了不同浓度肌原纤维蛋白对17种典型风味化合物吸附能力的影响。结果表明,风味化合物的种类对肌原纤维蛋白的吸附能力有显著影响,一定浓度的蛋白对醛、酮、醇、酯的吸附作用强弱依次为:醛类酯类酮类醇类。蛋白浓度由2 mg·m L~(-1)增加至4 mg·m L~(-1)时,对所有风味化合物吸附作用显著增强;由4 mg·m L~(-1)增加至6 mg·m L~(-1)时,蛋白对2-甲基丁醛、戊醛、己醛、丁酸乙酯吸附作用明显增强,而当蛋白浓度由6 mg·m L~(-1)增加至8 mg·m L~(-1)时,肌原纤维蛋白对所有风味化合物的吸附作用都明显减弱。蛋白对风味化合物的吸附能力的增加可能是由于较高的蛋白质浓度改变了风味化合物在气、液两相间的分配系数;然而过高浓度的蛋白对风味物质吸附能力的削弱则是蛋白质之间相互作用增强或表面张力降低的结果。本研究结果为肉类风味感知研究过程中确定蛋白浓度、挥发性风味化合物种类等关键参数和改善蛋白类食品的风味提供了参考。     

基于酶抑制法的农药残留快速比色检测

基于有机磷与氨基甲酸酯类农药对乙酰胆碱酯酶(acetylcholinesterase,AChE)的抑制作用,以吲哚酚乙酸酯为酶促水解反应的显色底物,制备了一种抛弃型农药残留快速检测酶片;通过优化试验,用物理吸附法将AChE固定到尼龙膜Hybond N+膜上,经真空冷冻干燥后,酶活回收率可达27.3%;该酶片显色结果为蓝绿色,通过比较显色强度的变化,酶片对农药标样氧乐果、毒死蜱、甲萘威与抗蚜威的检出限分别为1、0.05、1.5与0.8μg/mL,均达到食品中的最大残留限量标准;将研制的酶片用于葡萄汁与小白菜样品的检测结果证实,该酶片具有灵敏度高、准确度高、重现性好等优点,可用于农产品中有机磷与氨基甲酸酯类农药残留的快速定性筛检。     

酶的分子定向进化及其应用

酶的分子定向进化是20世纪90年代初兴起的一种蛋白质工程的新策略,是一种在生物体外模拟自然进化过程的、具有一定目的性的快速改造蛋白质的方法。该方法引起了生物催化技术领域的又一次革命。目前分子定向进化技术已被广泛应用于工业、农业及制药业等的相关领域。本文详细综述了酶的分子定向进化的概念、过程、基本策略及其核心技术,并着重介绍了酶的分子定向进化技术在提高酶的活力、稳定性、底物特异性和对映体选择性等几方面的应用及取得的相关成果。     

虎杖苷十一碳烯酸酯的酶法合成与动力学研究

为建立绿色、高效、高选择性合成虎杖苷十一碳烯酸酯双前药的生物催化体系,本研究以棉状嗜热丝孢菌(Thiermomyces lanuginosus)脂肪酶(TLL)为催化剂,以反应介质、底物摩尔比、温度及反应时间等因素为变量,探究了 TLL催化虎杖苷十一碳烯酰化反应的特性.结果表明,TLL可在丙酮中实现对虎杖苷的高效转化,...     

Elucidation of the enantioselective enzymatic hydrolysis of chiral herbicide dichlorprop methyl by chemical modification

Up to 25% of the current pesticides are chiral, the molecules have chiral centers, but most of them are used as racemates. In most cases, enantiomers of chiral pesticides have different fates in the environment. Knowledge of the function of amino acids of enzymes involved in enantioselective behaviors contributes to the understanding of the enantioselectivity of chiral pesticides. In this work, Aspergillus niger lipase (ANL, EC3.1.1.3) was chemically modified using bromoacetic acid (BrAc), 2,3-butanedione (BD), N-bromosuccinimide (NBS), and methanal. The enantioselectivity of the enzymatic hydrolysis of 2,4-dichlorprop-methyl (DCPPM) was investigated by chiral GC. The results have suggested that histidine, arginine, and tryptophan are essential for lipase activity and might be involved in the catalytic site of ANL. In addition, histidine and lysine play an important role in determining the observed enantioselective hydrolysis of chiral herbicide dichlorprop methyl. The molecular modeling study revealed that the essential hydrogen bonds formed between DCPPM and catalytic residues of ANL might be responsible for the enantioselectivity of DCPPM. The loss of enantioselectivity can also arise from the fact that the modification of the amino acids may cause changes in both the nature of the ANL enzyme conformation and the binding pattern of DCPPM. Our study provides basic information for the exploration of the enantioselective interaction mechanism of enzymes with chiral pesticides.     

Stereoselectivity of the generation of 3-mercaptohexanal and 3-mercaptohexanol by lipase-catalyzed hydrolysis of 3-acetylthioesters

The enantioselectivity of the generation of 3-mercaptohexanal and 3-mercaptohexanol, two potent sulfur-containing aroma compounds, by lipase-catalyzed hydrolysis of the corresponding 3-acetylthioesters was investigated. The stereochemical course of the kinetic resolutions was followed by capillary gas chromatography using modified cyclodextrins as chiral stationary phases. The enzyme preparations tested varied significantly in terms of activity and enantioselectivity (E). The highest E value (E = 36) was observed for the hydrolysis of 3-acetylthiohexanal catalyzed by lipase B from Candida antarctica resulting in (S)-configured thiol products. Immobilization of the enzyme (E = 85) and the use of tert-butyl alcohol as cosolvent (E = 49) improved the enantioselectivity. Modification of the acyl moiety of the substrate (3-benzoylthiohexanal) had no significant impact. The sulfur-containing compounds investigated possess attractive odor properties, and only one of the enantiomers exhibits the pleasant citrus type note.     

pH memory of immobilized lipase for (+/-)-menthol resolution in ionic liquid

Magnetic DEAE-GMA-EDMA microspheres were prepared via suspension polymerization and used for the immobilization of Candida rugosa lipase by ion exchange. The effect of pH values on the immobilization of lipase was investigated. Resolution of (+/-)-menthol in the hydrophobic ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate was performed by immobilized lipase-catalyzed enantioselective esterification with propionic anhydride as acyl donor. The effects of pH condition at lipase immobilization on the conversion and enantioselectivity were investigated. As a result, pH memory of the immobilized lipase for catalyzing (+/-)-menthol resolution in the ionic liquid was observed. Better conversion and the best enantioselectivity were obtained with the immobilized lipase prepared at pH 5.0. Under the condition, (-)-menthyl propionate with enantiomeric excess of >90% was obtained. Moreover, the enantioselectivity of the immobilized lipase decreased gradually with increasing pH value.     

Barley grain constituents, starch composition, and structure affect starch in vitro enzymatic hydrolysis

The relationship between starch physical properties and enzymatic hydrolysis was determined using ten different hulless barley genotypes with variable carbohydrate composition. The ten barley genotypes included one normal starch (CDC McGwire), three increased amylose starches (SH99250, SH99073, and SB94893), and six waxy starches (CDC Alamo, CDC Fibar, CDC Candle, Waxy Betzes, CDC Rattan, and SB94912). Total starch concentration positively influenced thousand grain weight (TGW) (r(2) = 0.70, p < 0.05). Increase in grain protein concentration was not only related to total starch concentration (r(2) = -0.80, p < 0.01) but also affected enzymatic hydrolysis of pure starch (r(2) = -0.67, p < 0.01). However, an increase in amylopectin unit chain length between DP 12-18 (F-II) was detrimental to starch concentration (r(2) = 0.46, p < 0.01). Amylose concentration influenced granule size distribution with increased amylose genotypes showing highly reduced volume percentage of very small C-granules (<5 μm diameter) and significantly increased (r(2) = 0.83, p < 0.01) medium sized B granules (5-15 μm diameter). Amylose affected smaller (F-I) and larger (F-III) amylopectin chains in opposite ways. Increased amylose concentration positively influenced the F-III (DP 19-36) fraction of longer DP amylopectin chains (DP 19-36) which was associated with resistant starch (RS) in meal and pure starch samples. The rate of starch hydrolysis was high in pure starch samples as compared to meal samples. Enzymatic hydrolysis rate both in meal and pure starch samples followed the order waxy > normal > increased amylose. Rapidly digestible starch (RDS) increased with a decrease in amylose concentration. Atomic force microscopy (AFM) analysis revealed a higher polydispersity index of amylose in CDC McGwire and increased amylose genotypes which could contribute to their reduced enzymatic hydrolysis, compared to waxy starch genotypes. Increased β-glucan and dietary fiber concentration also reduced the enzymatic hydrolysis of meal samples. An average linkage cluster analysis dendrogram revealed that variation in amylose concentration significantly (p < 0.01) influenced resistant starch concentration in meal and pure starch samples. RS is also associated with B-type granules (5-15 μm) and the amylopectin F-III (19-36 DP) fraction. In conclusion, the results suggest that barley genotype SH99250 with less decrease in grain weight in comparison to that of other increased amylose genotypes (SH99073 and SH94893) could be a promising genotype to develop cultivars with increased amylose grain starch without compromising grain weight and yield.     

非离子表面活性剂PEG对棉秆木质纤维素酶解的影响

为了提高纤维素的水解作用,减少酶的用量,该文研究不同分子量的聚乙二醇(PEG,poly ethylene glycol)表面活性剂对棉秆木质纤维素水解复合酶的影响,研究不同分子量的PEG对还原糖、纤维素酶、木质素酶、蛋白吸附率、酶动力学特征的影响。结果表明,2.5g/L添加量的PEG 2000、PEG 4000、PEG 6000和PEG 8000均能提高棉秆木质纤维素水解酶的活力,增加纤维素的转化率,减少蛋白的非特异性吸附率,提高棉秆木质素和纤维素的降解率,提高最大反应速度和降低酶与底物的亲和力,其中PEG 6000效果最好。3.0g/L的PEG 6000添加量对纤维素的转化率最高,达到58.12%(P<0.05)。对于吸附率,2.0、3.0、4.0、5.0g/L的添加量与对照相比差异显著(P<0.05),且相互之间差异不显著(P>0.05)。添加3.0g/L PEG 6000的表面活性剂能保持上清液中较高的蛋白浓度,水解3h后,没有添加表面活性剂的处理,纤维滤纸酶酶活从0.1245FPU/mL下降到0.012FPU/mL,下降了90.37%,添加PEG 6000后,FPA酶的活性恢复到0.041FPU/mL;水解48h,添加PEG 6000和不添加PEG 6000纤维素的转化率分别达到65.71%和54.68%,添加PEG 6000提高纤维素转化率20.18%。结果表明,PEG6000可以提高纤维素的转化率,为工业生产实践提供了参考。     

Adsorption and degradation of 2,4-dichlorophenoxyacetic acid in spiked soil with Fe0 nanoparticles supported by biochar

This work studies the adsorption and degradation of 2,4-dichlorophenoxyacetic (2,4-D) in spiked soil with nanoscale Fe⁰ particles (nFe⁰) and biochar derived from maize straw. When biochar concentration was high, the adsorption capacity of soil was enhanced. Furthermore, 2,4-D degraded completely at loading rates of 0.33 and 0.17 g/L nFe⁰ plus biochar (initial 2,4-D concentration of 10 mg/g) within 40 h, according to equilibrium data. Additionally, the theoretical concentration of chloridion was approximately 84%. Further analysis indicated that the effect of nFe⁰ on 2,4-D degradation was weaker in soil columns than that in soil slurry. By contrast, 2,4-D degradation is positively influenced by biochar application, which prevented the aggregation and corrosion of Fe nanoparticles. Although the enhanced capacity for 2,4-D adsorption on the soil decelerated dechlorination rate, long-term nFe⁰ activity was generated. After 72 h, the efficiency of 2,4-D degradation was approximately 53.2% in the soil columns with biochar support.     

Differences in Starch Granule Composition and Structure Influence In Vitro Enzymatic Hydrolysis of Grain Meal and Extracted Starch in Two Classes of Canadian Wheat (Triticum aestivum L.)

Starch granule composition and amylopectin structure affect starch digestibility, an important factor influencing wheat grain utilization for human food consumption. Six bread wheat cultivars with four belonging to the Canada Western Red Spring (CWRS) and two Canada Prairie Spring Red (CPSR) market classes were analyzed for the relationship between their grain constituents and in vitro enzymatic hydrolysis of starch. CPSR cultivars had higher starch and amylose concentrations compared with CWRS cultivars, which had a higher protein concentration. Starch granule size distribution did not differ among the genotypes, except AC Foremost, which had significantly (P < 0.05) higher volume percent of B‐type starch granules (≈15%) and lower volume percent of A‐type starch granules (≈9%) compared with other cultivars. Fluorophore‐assisted capillary electrophoresis revealed a lower content of R‐IV (DP 15–18, ≈6%) and a higher content of R‐VII (DP 37–45, ≈7%) chains in the CPSR cultivars compared with the CWRS cultivars. Starch in vitro enzymatic hydrolysis showed that compared with CWRS cultivars, the two CPSR cultivars had reduced amounts of readily digestible starch and higher amounts of slowly digestible starch and resistant starch. Consequently, the two CPSR cultivars also showed lower hydrolysis indexes in grain meal as well as extracted starch. CPSR cultivars, with higher starch and amylose concentrations, as well as a higher content of long chains of amylopectin, showed a reduced starch in vitro enzymatic hydrolysis rate.     

Protease Treatment for the Stabilization of Rice Bran: Effects on Lipase Activity,Antioxidants, and Lipid Stability

Rice bran lipid is rapidly made rancid by endogenous lipase enzymes. To inactivate rice bran lipase, an enzymatic hydrolytic method was developed and then compared with the thermal method. The efficiency of five proteolytic enzymes including trypsin, chymotrypsin, papain, bromelain, and Flavourzyme enzyme to stabilize rice bran was investigated. Moreover, the antioxidant content and storage stability of enzymatically stabilized rice bran (ESRB), thermally stabilized rice bran (TSRB), and raw rice bran (RRB) were studied. Trypsin, chymotrypsin, and papain showed a higher rate of hydrolysis than bromelain and Flavourzyme enzyme. After 120 min of hydrolysis, rice bran hydrolyzed by trypsin, chymotrypsin, and papain inactivated 80, 86, and 79% of lipase activity, respectively, whereas lipase activity of rice bran hydrolyzed by bromelain and Flavourzyme enzyme was higher than the initial rice bran. With a similar lipase inactivation level, the cheaper papain was used to produce ESRB. Total phenolics content of ESRB was 52.89 and 94.10% higher than in RRB and TSRB, respectively. In addition, γ‐oryzanol content in ESRB was 2.23‐ and 2.05‐fold of that in RRB and TSRB, respectively. Lipase activity of RRB increased throughout the two months of storage, whereas no change in lipase activity was observed in ESRB and TSRB. At the end of storage, free fatty acid contents of RRB, TSRB, and ESRB were 15.30, 4.67, and 3.92%, respectively. We propose enzymatic hydrolysis by papain for stabilization of rice bran with high antioxidant content and storage stability.     

Limited enzymatic hydrolysis can improve the interfacial and foaming characteristics of beta-conglycinin

In this contribution, we have determined the effect of limited enzymatic hydrolysis on the interfacial (dynamics of adsorption and surface dilatational properties) and foaming (foam formation and stabilization) characteristics of a soy globulin (beta-conglycinin, fraction 7S). The degree of hydrolysis (DH = 0, 2, and 5%), the pH of the aqueous solution (pH = 5 and 7), and the protein concentration in solution (at 0.1, 0.5, and 1 wt %) were the variables studied. The temperature and the ionic strength were maintained constant at 20 degrees C and 0.05 M, respectively. The rate of adsorption and surface dilatational properties (surface dilatational modulus, E, and loss angle) of beta-conglycinin at the air-water interface depend on the pH and DH. The adsorption decreased drastically at pH 5.0, close to the isoelectric point of beta-conglycinin, because of the existence of a lag period and a low rate of diffusion. The interfacial characteristics of beta-conglycinin are much improved by enzymatic treatment, especially in the case of acidic aqueous solutions. Hydrolysates with a low DH have improved functional properties (mainly foaming capacity and foam stability), especially at pH values close to the isoelectric point (pI), because the protein is more difficult to convert into a film at fluid interfaces at pH approximately equal to pI.     

Enzymatic and Acidic Hydrolysis of Cationized Waxy Maize Starch Granules

A series of wet‐cationized starch granules from waxy maize with different degrees of substitution (DS) were solubilized with either 2.2M HCl (lintnerization) or with the α‐amylase of Bacillus amyloliquefaciens. The maximum rate of the enzymatic hydrolysis occurred in starches with intermediate DS. It appeared that the cationic substituents interfered with the binding to the active site of the enzyme at high levels of substitution. The DS remained fairly constant in the granular residues after the enzymatic attack. The rate of the acidic hydrolysis increased with increasing DS but the final level of solubilization slightly decreased. The DS of the residual starch material decreased to 40% of the original level, showing that a large part of the cationic groups was found within the amorphous parts of the granules. A dry‐cationized sample with a high DS was also treated with the acid and lost a major part of its substituents at low levels of lintnerization. Probably most of the substituents were associated with the surface and channels of these granules. The cationized starches possessed branches that were resistant to isoamylase attack and the samples also contained β‐amylolysis resistant dextrins. The proportion of resistant dextrins in the granular residues decreased after lintnerization, but remained constant after the enzymatic hydrolysis.