采用双层静电自组装技术制备nPAM-木瓜蛋白酶-CNC催化剂

本研究利用载体材料纤维素纳米晶(CNC)、木瓜蛋白酶(papain,PA)以及非离子型聚丙烯酰胺(non-ionic polyacrylamide,n PAM)的表面电荷特性,制备了新型的固定化木瓜蛋白酶,利用Zeta电位分析,探寻固定化过程中的静电作用机制;并通过傅立叶变换红外光谱(FT-IR)分析固定化PA的组成成分和各物质的相互作用。通过对固定化过程中的各因素进行研究,得到了制备该固定化酶的最优条件,分别为p H 5.0,CNC和PA质量比为36:1(CNC/PA),n PAM(5%质量分数)的添加量为0.3 m L,酶层自组装时间30 min;在此条件下酶活回收率达85.8%。随后,对游离酶和固定化酶的动力学参数研究显示固定化酶的Vmax/Km值(1.40 min-1)明显高于游离酶(0.80 min-1),表明所制备的固定化PA具有更高的催化效率、更广的最适p H范围和温度范围以及操作稳定性,在重复使用5批次之后,相对酶活仍在96.5%以上。可见双层自组装技术是一种进行固定化酶的新途径。     

Mimicking Spray Drying by Drying of Single Droplets Deposited on a Flat Surface

The inactivation of bioactive ingredients during spray drying is often matrix specific. Therefore, the design of new processes or the optimisation of existing spray drying processes is usually highly product specific and requires numerous experiments. Rapid experimentation methods that facilitate fast data generation are therefore desired. A novel method for drying single droplets to mimic spray drying is proposed. The approach involves droplet deposition on a hydrophobic flat surface followed by controlled drying. A heat and mass transfer model is applied to predict the drying history of the single droplets. The approach is successfully evaluated through studying the inactivation of β-galactosidase during drying. The heat and mass transfer model supplemented with inactivation kinetics provided reasonable prediction of the residual enzyme activity after drying. In addition, the inactivation kinetics could be directly extracted from single droplet experiments rather than using the kinetics from separate heating experiments. Finally, it was demonstrated that the inactivation kinetics found with the single-drop experiments could satisfactorily predict the residual activity of β-galactosidase dried with a laboratory-scale spray dryer.     

Variations of peroxidase activity in cocoa (Theobroma cacao L.) beans during their ripening,fermentation and drying

An increase of peroxidase activity in the seeds of cocoa (Theobroma cacao L.) during their ripening has been determined. An additional increase of the peroxidase activity (about 10 times) is observed during the fermentation and drying of the beans. The residual activity determined in the cocoa beans after sun-drying was higher than that in unfermented ripe seeds. The major cocoa isoperoxidase was shown to be an acidic enzyme with pI 4.7. Using isoelectrofocusing, the appearance of two basic isoenzymes of the peroxidase with pI 8.6 and 9.0 during the process of the fermentation has been detected. ©     

啤酒酵母乙醛代谢关键酶相关性研究

乙醛是啤酒中的主要风味物质,其代谢主要来自酵母细胞。酵母中乙醇脱氢酶及乙醛脱氢酶是乙醛代谢的关键酶,对乙醛变化起着重要作用。跟踪啤酒酵母发酵过程中相对酶活力及乙醛变化,发现两种乙醇脱氢酶和乙醛脱氢酶的相对酶活力与发酵过程乙醛含量变化具有一定相关性。同时对低产乙醛啤酒酿酒酵母kb2-4与出发菌株啤酒酵母kb进行发酵试验,跟踪检测相对酶活力及乙醛含量,其乙醇脱氢酶Ⅰ和乙醇脱氢酶Ⅱ及乙醛脱氢酶相对酶活力均高于出发菌株,平均增幅分别为15.5%,11.6%和5%。3种酶活性的变化协同作用可以使乙醛含量降幅最大为33.8%。     

盐渍对鲍鱼微波真空干燥过程和品质的影响

盐渍是鲍鱼微波真空干燥过程前处理的关键因素. 以不同浓度盐溶液-0, 5%, 7 5%, 10%, 15%和饱和盐溶液-盐渍鲍鱼,在微波功率2 000 W、真空度-80 kPa条件下进行微波真空干燥. 以其微波真空干燥特性、干燥后色泽、复水性及复水后质构特性为指标,考察盐渍对鲍鱼干燥过程和品质的影响. 结果表明：蒸馏水浸泡和盐浓度大于15%的盐溶液盐渍都不利于鲍鱼微波真空干燥的进行和干制后的品质. 当食盐浓度为7 5%~10%时,鲍鱼的微波真空干燥速率明显加快,复水率和复水比均较好,复水后呈现较佳咀嚼性,且色泽为均匀的淡黄色. 综合考虑,经过7 5%盐溶液浸渍24 h后,沸水煮2 min的鲍鱼进行微波真空干燥效果较佳.     

Effect of temperature cycling on allinase activity in garlic

Allinase, which catalyzes the conversion of alliin to allicin, the principal component of potential medicinal value in garlic, is a thermo-labile enzyme. The potential for allicin formation is determined by the quantity of allinase that remains active after the process of preserving garlic by drying. The kinetics of enzymatic activity loss during drying by temperature cycling or by constant temperature were evaluated and compared. Allicin-forming potential was 91% preserved by temperature cycling from 40 to 60 °C. It was found that sugars present in the garlic and the high molecular mass of the enzyme were responsible for protection against degradation at high drying temperatures. Preservation of the enzymatic activity under cyclical conditions occurred mainly with exposure to low temperatures for drying periods longer than those of constant drying conditions.     

固定化Bacillus thuringiensis ZJOU-010壳聚糖酶的研究

采用吸附交联技术,以DEAE-22 纤维素为载体、戊二醛为交联剂,固定Bacillus thuringiensis ZJOU-010壳聚糖酶,考察固定化酶的制备条件,并研究固定化酶的性质。结果表明B.thuringiensis ZJOU-010 壳聚糖酶的最佳固定化条件为：戊二醛体积分数3.0%、加酶量20mg、固定化时间10h;在此条件下制备的固定化壳聚糖酶的最适pH 值和温度分别为4.83 和50℃;与游离酶相比,该固定化酶的热稳定性较好,在40℃和50℃条件下的半衰期(t1/2)分别为36.3h 和6.2h,动力学常数Km 值为9.19g/L;该固定化酶重复使用10 批后活力仍可保持初始活力的88.32%。     

黑曲霉N5-5产单宁酶的酶学性质与固定化

本实验室自主分离的黑曲霉N5-5所产单宁酶已发现对没食子酸丙酯具有良好酶解效果。为了单宁酶的工业化应用,本次研究大批量培养单宁酶,探究其对单宁酸的酶解效果及酶的固定化效果。发酵采用先液体扩培后固体发酵的形式,提酶后利用陶瓷膜过滤技术纯化、浓缩酶液,并对比冷冻干燥和喷雾干燥两种不同干燥方式,还使用树脂载体对酶进行固定化。实验表明,纯化后单宁酶酶活达258.53 U/mL,可水解10%至50%浓度的单宁酸,其中30%以下底物浓度酶解效果较好;冷冻干燥对酶活影响不大而喷雾干燥使酶活降为174.02 U/mL;另外,单宁酶通过树脂载体固定化后,在实验条件下可重复使用至少4次。研究为提高黑曲霉N5-5发酵所产单宁酶的媒介效率、降低酶解反应成本、实现商业化生产建立了理论基础。     

Activity of diastase α-amylase immobilized on polyanilines (PANIs)

Starch hydrolysing enzyme α-amylase (EC 3.2.1.1) was immobilized by physical adsorption and covalent binding onto chemically synthesised polymer, polyanilines (PANIs) in two different forms, emeraldine salt and emeraldine base powder. The immobilization efficiency was affected by the pH of the immobilization medium, contact time and amount of enzyme. The kinetic parameters, reusability and storage stability of the enzyme were studied under free and immobilized condition at optimum pH and temperature. The immobilized enzyme showed enhanced storage stability over a period of 6 months, while free enzyme in solution lost all of its activity within a period of 8 days. Reusability of the enzyme was improved by immobilization. The K_m values for starch hydrolysis were found to be high for the immobilized enzymes.     

Activity of cathepsin B, D, H and L in Spanish dry-cured ham of normal and defective texture

Thirty-six specimens of Spanish dry-cured ham from 18 different batches were studied to determine the influence of endogenous enzymes on the textural quality of the end product. It was shown that, the residual enzyme activity of cathepsins B+L is a reliable indicator of textural defects associated with strong proteolysis in Spanish dry-cured ham. In contrast, the activity of cathepsin B, which varied widely among samples, was less influential. Salt levels can have a moderate effect on the residual enzyme activities. Other compositional parameters also influence the properties of the end product; the amount of protein in semimembranosus muscle was found to be less and the moisture greater in texturally defective specimens. Also, the efficiency of the drying-curing process was less in hams yielding poor textures.     

Effects of drying methods and conditions on release characteristics of edible chitosan films enriched with Indian gooseberry extract

The present work was aimed at studying the effects of drying methods and conditions (i.e., ambient drying, hot air drying at 40 °C, vacuum drying and low-pressure superheated steam drying within the temperature range of 70–90 °C at an absolute pressure of 10 kPa), as well as the concentration of Indian gooseberry extract, (added to edible chitosan film-forming solution as a natural antioxidant, at concentrations of 1, 2 and 3/100 g), on the residual total phenolic content (TPC) of the films. The swelling and release behaviour of TPC from the films were also studied. Drying methods and conditions were found to have significant effects on the percentage of residual TPC. The release characteristics, swelling and functional group interaction of the antioxidant films, as assessed by Fourier-transform infrared (FTIR) spectroscopy, were found to be affected by the drying methods and conditions, as well as the concentration of the Indian gooseberry extract.     

EFFECTS OF HARVESTING MATURITY AND OFF-PLANT RIPENING ON THE ACTIVITIES OF LIPOXYGENASE, HYDROPEROXIDE LYASE, AND ALCOHOL DEHYDROGENASE ENZYMES IN FRESH TOMATO

The specific activities of lipoxygenase (LOX), hydroperoxide lyase (HPL), and alcohol dehydrogenase (ADH) were determined at green, pink, and red stages of ripeness in twelve tomato cultivars ripened during storage. Also, the same measurements were done in six other cultivars harvested at breaker (BR) and red (RR), and ripened to full‐red stage of maturity. LOX and HPL showed highest activities at the pink stage of ripeness, whereas ADH was most active at the red stage. Variability of enzyme activity among individual fruit and of hue angle within ripeness stage was great. There were differences among the twelve lines for each enzyme activity, suggesting the potential for altering tomato flavor by selecting lines with different enzyme patterns. Harvest maturity was not a factor in the activities of LOX and HPL, but effective in ADH activity. In almost all cultivars, ADH activity was found to be higher in red harvested counterparts.     

Properties of Rhodotorula gracilis D-amino acid oxidase immobilized on magnetic beads through his-tag

D-amino acid oxidase catalyzes one of the key steps in the production of semisynthetic cephalosporins. We expressed and purified recombinant Rhodotorula gracilis D-amino acid oxidase with C-terminal his-tags. This engineered enzyme was immobilized onto Ni(2+)-chelated nitrilotriacetic acid magnetic beads through the interaction between his-tag and Ni(2+). The kinetic constants, storage properties, and the reusability of the immobilized d-amino acid oxidase were determined. The effects of temperature, pH, and hydrogen peroxide on the activity of immobilized d-amino acid oxidase were also studied. The highest activity recovery was 75%. Thermal stability was improved after immobilization; the relative activity of the immobilized enzyme was 56% whereas the free enzyme was completely inactivated after incubation at 50 degrees C for 1 h. In the presence of 10 mM hydrogen peroxide, the immobilized enzyme did not show a rapid loss of activity during the first 2 h of incubation, which was observed in the case of the free enzyme; the residual activity of the immobilized enzyme after 9 h was 72% compared with 22% of the free form. The long-term storage stability was improved; the residual activity of the immobilized enzyme was 74% compared with 20% of the free enzyme when stored at room temperature for 10 d. The immobilized form retained 37% of its initial activity after 20 consecutive reaction cycles.     

Dehydrated Blueberries by the Continuous Explosion-Puffing Process

A blueberry dehydration process which includes the unique continuous explosion-puffing system (CEPS) is described. A drying study including alternate drying pretreatments failed to increase the dehydration rate. It was found that care during processing was necessary to prevent rupture of the berries as rupture caused bleeding during drying which reduced the drying rate. Optimum operating conditions for CEPS were established for blueberries. Measurements of dried blueberry properties such as bulk density, color, rehydration, and disintegration were used to determine optimum operating conditions for pressure (103 kPa), temperature (190°C), and feed moisture (18.5%) for CEPS.     

板栗微波真空干燥特性及干燥工艺研究

研究微波真空干燥方式下,微波强度、腔体压力等参数对板栗干燥过程中质热传递的影响规律。采用Box-Behnken中心组合试验设计,以水分含量和白度值为评价指标,确定板栗微波真空干燥过程中微波功率、压力、干燥时间的最适工艺参数。结果显示:板栗微波真空干燥过程主要为加速和降速阶段,恒速阶段持续时间较短。微波功率和真空度均对干燥时间有显著影响,功率越大,真空度越高,干燥速率越快,干基含水率和水分比都随着干燥时间的延长而逐渐下降。由回归模型得出板栗微波真空干燥的最佳工艺参数为时间12min,压力-56kPa,功率3kW。微波真空干燥的微波功率、腔体压力、干燥时间均对板栗品质有影响,以模型得出的干燥参数进行干燥,可保证板栗干燥后的品质,且干燥效率高、能耗低。     

ECR-1030M树脂固定化单宁酶

制备了一种新型固定化单宁酶并优化了固定化工艺。用米曲霉固态发酵单宁酶,然后分别用不同类型树脂以戊二醛交联法固定单宁酶并筛选出最佳树脂载体;通过均匀试验和统计分析优化了单宁酶固定化工艺条件;对固定化单宁酶的保藏稳定性和循环使用次数进行了测定。结果显示:二苯乙烯/丙烯酸酯基型大孔吸附树脂ECR-1030M固定化单宁酶性能最佳。最优固定化工艺条件为:戊二醛浓度3.5%、固定化时间5.5 h、料酶比1︰16(g/m L)。此条件下制备的固定化单宁酶活力最高,为1480.5±32.6 U/g,酶活回收率为64.5%。固定化单宁酶在4℃下保藏20 d,以及循环使用6次后,仍可保留超过80%的酶活力。由此可见,ECR-1030M树脂固定化单宁酶是一种酶活力高、稳定性好、可多次高效使用的酶制剂。     

壳聚糖-埃洛石纳米管微球的制备及其对木瓜蛋白酶的固定化

运用交联-吸附法制备壳聚糖-埃洛石纳米管(chitosan-halloysites nanotube,CTS-HNTs)复合微球固定木瓜蛋白酶,并通过傅里叶红外光谱、扫描电子显微镜、荧光标记等方法进行表征。2%CTS+1%HNTs制备的微球对木瓜蛋白酶的固定量最高。固定化条件为木瓜蛋白酶质量浓度1 mg/mL、固定化时间10 h。固定化木瓜蛋白酶最适pH 6.8(游离酶pH 7.2)、最适温度60℃(游离酶50℃),保存30 d该酶相对活性为62%(游离酶27%),使用4次后,木瓜蛋白酶的相对活性仍然保留26.26%。CTS-HNTs微球固定化木瓜蛋白酶耐贮存,操作稳定性强,可以提高酶的利用率,降低酶解的成本,提高生产效率。     

巴氏醋酸杆菌AS1.41产醋酸关键酶研究

巴氏醋酸杆菌（Acetobacer pasteurianus）将乙醇氧化成醋酸的关键酶是乙醇脱氢酶（ADH）和乙醛脱氢酶（ALDH）。在不同初始乙醇含量条件下,ADH和ALDH的酶活呈现动态变化,乙醇含量为4%时,ADH和ALDH的酶活达到最大,分别为7.43 U/mg和7.18 U/mg。同时,酶活与产酸速率呈现出较高一致性：酶活越高,产酸速率越快。发酵温度为32 ℃时,菌体生长最为活跃,酶活最大,产酸最快;加入0.5%的乙酸后,ADH和ALDH的酶活分别由8.12 U/mg和7.06 U/mg提高到了9.43 U/mg和8.52 U/mg,产酸速率也得到相应提升。ALDH对乙醇、乙酸、温度的稳定性均高于ADH。     

Effect of simultaneous infrared dry-blanching and dehydration on quality characteristics of carrot slices

This study investigated the effects of various processing parameters on carrot slices exposed to infrared (IR) radiation heating for achieving simultaneous infrared dry-blanching and dehydration (SIRDBD). The investigated parameters were product surface temperature, slice thickness and processing time. A three-factor factorial design was conducted to determine the influence of processing parameters on moisture reduction, drying rate, residual peroxidase (POD) activities, surface color change and vitamin C retention. High surface temperature and/or thin slices had faster inactivation of enzymes and quicker moisture removal compared to the low surface temperature and/or thick slices. A Middili model performed well for describing drying behavior during the treatment, while a biphasic model and a quantic model fit well for describing the POD inactivation and the surface color change during the process, respectively. The process which produced 1 log reduction in POD activity has resulted in moisture reduction from 40.2 to 88.8 g/100 g, overall color change (ΔE) from 3.17 to 5.13 and retention of vitamin C from 56.92 to 77.34 g/100 g compared to control. It was concluded that SIRDBD could be used as an alternative to produce high quality blanched and partially dehydrated fruits and vegetables.     

响应曲面法漂白紫胶真空干燥条件的优化

为得到漂白紫胶真空干燥的优化条件,研究了不同温度和压强条件下漂白紫胶产品的颜色指数和干燥时间,用响应曲面法(RSM)设计试验并建立了真空干燥方法的拟合方程。结果表明,预测值和实测值的一致性较好,所建立的数学模型可以用于描述漂白紫胶的真空干燥。通过优化得到了漂白紫胶真空干燥的最优条件：温度30℃,压强30kPa,可以得到颜色指数为1.0 的漂白紫胶,其指标符合国家精制漂白紫胶1 级标准;温度50℃,压强30kPa,可以得到颜色指数为1.4 的漂白紫胶,其指标符合国家精制漂白紫胶2 级标准。同时对此条件进行了验证,结果表明这两种条件下制得的漂白紫胶符合国家标准,可以为生产提供参考。