Effect of ultrasound and enzymatic pre-treatment on yield and properties of banana juice

Effect of ultrasound and enzymatic pre-treatments with cellulase and pectinase on yield and properties of banana juice were investigated. A two-level full factorial design was employed. The factors selected were ultrasonication time (0 and 30 min), cellulase concentration (0 and 0.2%) and pectinase concentration (0 and 0.2%). The responses studied were yield, viscosity, clarity, total soluble solids (TSS) and pH. It was observed that pectinase was more effective in increasing the yield of juice compared to cellulase. Ultrasonic pre-treatment alone did not significantly increase the yield of juice. When ultrasound was combined with pre-treatment with both the enzymes maximum yield of 89.40% was obtained compared to 47.30% in the control. The viscosity of the juice decreased with addition of enzymes and with application of ultrasound. The clarity of the juice was not affected by cellulase treatment, but improved with pectinase treatment. Ultrasonication alone was found to be more effective than pectinase or cellulase treatment in improving the clarity of the juice. The TSS increased with enzymatic treatment, ultrasonication and their combination. pH was not affected by treatment type, but was found to be lower for the treated juices. Significant correlations were observed between the various responses.     

氦-氖激光诱变细菌细胞及原生质体选育果胶酶高产菌株

利用氦-氖激光(波长632.8nm,功率15mW)诱变果胶酶产生菌的菌体细胞,获得了突变株ZH-g,其酶活达到301 u·mL－1,是出发菌株的1.3倍.进一步用氦-氖激光照射ZH-g的原生质体,从而得到了高产突变株ZH-gA,其酶活达到357 u·mL－1,比ZH-g提高了18%,是出发菌株的1.6倍.该菌株在好氧和静置条件下均能良好产酶,经传代实验证明其产酶性能稳定.     

The effect of pectinase on the bubble fractionation of invertase from α-amylase

Fermentation broth normally contains many extracellular enzymes of industrial interest. To separate such enzymes on-line could be useful in reducing the cost of recovery as well as in keeping their yield at a maximum level by minimizing enzyme degradation from broth proteases (either the desired enzymes or the proteases could be removed selectively or both removed together and then separated). Several large-scale separation methods are candidates for such on-line recovery such as ultrafiltration, precipitation, and two-phase partitioning. Another promising technique for on-line recovery is adsorptive bubble fractionation, the subject of this study. Bubble fractionation, like ultrafiltration, does not require contaminating additives and can complement ultrafiltration by preconcentrating the enzymes using the gases normally present in a fermentation process. A mixture of enzymes in an aqueous bubble solution can, in principle, be separated by adjusting the pH of that solution to the isoelectric point (pI) of each enzyme as long as the enzymes have different pIs. The model system investigated here is comprised of three enzyme separations and the problem is posed as the effect of pectinase (a charged enzyme) on the bubble fractionation of invertase (a relatively hydrophilic enzyme) from α-amylase (a relatively hydrophobic enzyme). The primary environmental variable studied, therefore, is the pH in the batch bubble fractionation column. Air was used as the carrier gas. This prototype mixture exemplifies an aerobic fungal fermentation process for producing enzymes. The enzyme concentration here is measured as total protein concentration by the Coomassie Blue (Bradford) solution method (1), both as a function of time and column position for each batch run. Since, from a previous study (2), it was found that invertase and α-amylase in a two-enzyme system can be partially separated in favor of one vs the other at two different pHs (pH 5.0 and 9.0) with significant separation ratios, emphasis is placed on the effect of pectinase at these pHs. In this study, the addition of pectinase reduced the total separation ratio of the α-amylase-invertase mixture at both pHs.     

紫外、氦氖激光等复合诱变产果胶酶细菌ZH1的研究

ZH₁菌株是从自然界分离筛选而来的,该菌株产果胶酶的最适pH为7.0;最适温度33℃;培养36h达产酶高峰,酶活力为73.5μ/mL,将ZH₁作为出发菌株,经紫外线诱变、硫酸二乙酯诱变,亚硝基胍和紫外线复合诱变及氦氖激光等多次反复诱变,选育得到一株产果胶酶性稳定且酶活明显提高的突变株ZHg,其酶活为301m/mL,比出发菌株ZH₁产果胶酶能力提高3.1倍.     

Magnetite Nanoparticles Immobilized Pectinase: Preparation,Characterization and Application for the Fruit Juices Clarification

In this work, pectinase was immobilized on the surface of silica‐coated magnetite nanoparticles via covalent attachment. The magnetite‐immobilized enzyme was characterized by Fourier transform infrared spectroscopy, X‐ray powder diffraction, scanning electron microscopy and vibrating sample magnetometery techniques. Response Surface Methodology using Minitab Software was applied for statistical designing of operating conditions in order to immobilize pectinase on magnetic nanoparticles. The optimal conditions were obtained at 30 °C and pH 5.5 with 42.97 μl pectinase for 2 h. The immobilization yield was 50.6% at optimized conditions. Compared to the free pectinase, the immobilized pectinase was found to exhibit enhanced enzyme activity, better tolerance to the variation of pH and temperature, and improved storage stability. Both free and immobilized samples reduced the viscosity of apple juice from 1.12 to 0.88 and 0.92 mm²s^?1, respectively, after 30 min at their optimum temperature. Furthermore, the immobilized enzyme could be reused six consecutive cycles and the efficiency loss in viscosity reduction was found to be only 8.16%.     

Laboratory tests on glycosidase preparations in wine

The secondary glycosidasic activity of pectinase preparations is used to release flavour active compounds of wine. In order to identify any undesirable effects of other collateral enzymatic activities, laboratory tests were carried out using two commercial preparations: NOVAROM G (Novo-Nordisk) and AR 2000 (Gist-Brocades). A pool of wines was obtained by blending two white wines from neutral grapes (Chardonnay and Verdeca) and a red wine from aromatic grapes (Aleatico). The experiment was carried out by adding each enzymatic preparation to the sample and analysing, by gas chromatography, with flame ionisation and mass spectrometric detection, the free aroma compounds after 15 and 30 days. In the first 15 days, the glycosidasic activity of the NOVAROM G brought about a considerable increase in the wine’s free terpenic content, especially nerol, geraniol and citronellol. After 30 days, further increases in all terpenic compounds were observed. The AR 2000, after 15 days, brought about the release of a conspicuous amount of terpenes, more than the previous enzymatic preparation, which further increased after 30 days. Throughout the test period, the enzymatic preparation, NOVAROM G, had no effect on the content of the acetates of the higher alcohols, the compounds associated with the banana-pineapple (isoamyl acetate) and rose (2-phenyl ethyl acetate) scents. However, the AR 2000 brought about a rapid hydrolysis of total acetates, which caused a fifty percent reduction in isoamyl acetate after 15 days and continued to decline until, after 30 days, the remaining concentration was below the perception threshold. Neither of the enzymatic preparations, however, provoked significant effects on the ethyl esters of the fatty acids compared to the control wine. In addition, the AR 2000 revealed esterase activity involving fixed acids, such as malic acid and succinic acid, with a conspicuous formation of diethyl malate and diethyl succinate, which became more marked after 30 days.     

Overproduction of Laccase and Pectinase by Microbial Associations in Solid Substrate Fermentation

The growth and the enzymatic production of two microbial fungal associations were studied: Aspergillus niger and Fusarium moniliforme and Trametes versicolor and Aspergillus niger. The synergistic interrelations between the species of the first mixed culture increased the biosynthesis of α-amylase and pectinase. T. versicolor and A. niger proved to be compatible partners in the overproduction of the enzyme laccase, whose synthesis surpassed 8.4 times the enzymatic level in the monoculture, with both of the mixed microbial populations cocultivation facilitating the amplified synthesis of enzymes rather than their growth acceleration. A further proof of the presence of synergism established by the cultures was the enzyme volumetric productivities in both of the mixed microbial cultures, which increased parallel to the rise in the combined biomass synthesis. The competent selection of compatible partners can adjust the desired enzymatic levels and compositions in mixed fungal systems aimed at a number of specified designations. Thus, a very high level of laccase production (97,600 IU/g dry weight) was achieved. The chosen fungal strains produce a variety of different enzymes, but first microbial association produces mainly amylase and pectinase, necessary for their growth, and second association produces mainly laccase and pectinase.     

Enzymatic hydrolysis of pectic substances by gel-permeation chromatography

Summary A rapid liquid chromatographic (HPLC) method for the determination of pectinolytic activity of enzymes produced by Aspergillus Niger and Rhizopus species is reported.Compared with more conventional methods, HPLC is more reliable and has a much improved maximum sensitivity. The limit of quantitation of galacturonic acid is 0.1g.     

激光诱变选育果胶酶高产菌株

采用Ar⁺、He-Ne、LD、YAP等激光辐照果胶酶产生菌,不同波长激光辐照均获高酶活菌株,其中从LD激光与He-Ne激光辐照组中选育出酶活达6×10⁴单位/g的高产菌株(比对照组提高122%),并稳定地应用于生产.文中从黑曲霉抱子与DNA的吸收光谱以及电镜观察黑曲霉抱子的结果出发,对激光诱变机制进行了初步探讨.     

激光诱变选育果胶酶高产菌株

采用Ar＋、He－Ne、LD、YAP等激光辐照果胶酶产生菌，不同波长激光辐照均获高酶活菌株，其中从LD激光与He－Ne激光辐照组中选育出酶活达6×104单位／g的高产菌株（比对照组提高122％），并稳定地应用于生产．文中从黑曲霉抱子与DNA的吸收光谱以及电镜观察黑曲霉抱子的结果出发，对激光诱变机制进行了初步探讨．