超滤法浓缩菊糖果糖转移酶的初步研究

超滤浓缩的菊糖果糖转移酶粗酶根据用途可直接使用或进一步纯化,采用截留分子量为10kDa的Pellicon-2盒式超滤膜包,对菊糖果糖转移酶超滤浓缩进行了初步研究。结果表明,酶解菊糖产物为DFAIII;酶收率85%以上,膜通量减少至3.2L/m2·h;单位体积酶活(U/mL)增加8.5倍,比酶活达到6.7U/mg;再生后,膜通量可以恢复95%以上。     

β-葡萄糖苷酶粗酶液的超滤条件研究

采用超滤技术分离纯化β-葡萄糖苷酶粗酶液,以膜通量、回收率和比酶活为综合评价指标,筛选出合适的截留分子量的膜材料,并进一步考察了进口压力、回流压力、粗酶液固形物含量、膜过滤时间等因素对β-葡萄糖苷酶粗酶液超滤浓缩的影响。结果表明,β-葡萄糖苷酶粗酶液最佳超滤工艺条件为:选用50 ku的超滤膜,对0.9%的料液浓度在进口压0.12 MPa、回流压0.08 MPa条件下超滤20 min,浓缩倍数达3倍,β-葡萄糖苷酶回收率达95.05%,比酶活提高1.24倍,平均膜通量达96.0 L/(m~2·h)。用超滤膜浓缩β-葡萄糖苷酶具有工艺简单、能耗低、绿色环保、β-葡萄糖苷酶活性损失小等优点,为β-葡萄糖苷酶的工业化生产放大奠定基础。     

应用膜分离技术改进泰乐菌素提取工艺

应用超滤-纳滤组合膜分离技术对泰乐菌素发酵液进行提纯和浓缩,以取代原有的板框过滤。对不同材料、截留分子量的超滤和纳滤膜的筛选结果表明,超滤膜宜采用PES-150聚醚砜膜,纳滤膜宜采用聚酰胺复合纳滤膜DL 2540。发酵液采用硅藻土预处理,且超滤过程恒容累积加水250L,能提高82%的膜通量,此时泰乐菌素的收率达97%。发酵液超滤液与原板框滤液的泰乐菌素组分纯度基本一致,但超滤液萃取效果更好,萃取收率提高了20%～26%。DL 2540纳滤膜对超滤液进行3.5倍浓缩,平均通量可达41.3 L/(m2.h),通过纳滤收率可达99%,纳滤浓缩液萃取无乳化现象,所得萃取液的色度也较低。     

枯草芽孢杆菌氨肽酶超滤提取技术

采用Ultra-flo超滤系统对经离心除菌、质量分数18%的(NH4)2SO4澄清的发酵液进行超滤浓缩和脱盐,为获得该过程中的高酶活回收率和膜通量,对其工艺条件进行了研究和优化;选用截留分子量为30 kDa的聚醚砜(PES)膜,操作压力为0.2～0.25 MPa,澄清发酵液初始pH值在5.5～6.5,浓缩倍数3倍;采用定时流加水的方式超滤脱盐,流加水体积控制为浓缩发酵液体积的10倍。在此优化条件下,超滤浓缩过程的酶活回收率达85%,超滤脱盐过程酶活回收率达79%,该过程可除去90%以上的盐,终浓缩倍数可控制为8～10倍;整个超滤过程的酶活回收率为67.15%,并且整个过程中膜通量维持在16 L/(m2.h)以上。     

超滤法提取发酵液中核黄素

采用截留分子质量(MWCO)为1000 Da、10 kDa和30 kDa的卷式膜,探索卷式膜超滤核黄素发酵液的可行性。对比分析超滤膜的杂质截留率和膜通量,发现MWCO为10 kDa的卷式超滤膜对核黄素具有较好的分离效果。通过研究操作压力、温度、流速及浓缩倍数对膜通量的影响,确定核黄素发酵液超滤的最佳条件为:温度30℃,流速5 m/s,操作压力0.2 MPa,浓缩倍数为3.8倍。超滤后,经过氧化结晶核黄素的纯度可达98.6%,总回收率达95%。     

超滤和反渗透技术在果汁生产中的应用

根据超滤膜和反渗透膜的特点，介绍了超滤(UF)和反渗透(RO)膜技术在苹果汁、橘子汁、山楂汁等果汁生产中的应用。     

膜浓缩技术制备黑果枸杞花青素的工艺研究

以黑果枸杞干果为原料制备黑果枸杞花青素浓缩汁,分析了影响黑果枸杞提取汁质量的因素。采用超滤、反渗透膜浓缩技术,根据浓缩前后黑果枸杞花青素及可溶性固形物含量的变化规律,确定最佳工艺条件。结果显示:最佳干果复水比为1∶6(g/mL);复水后,经酶解的黑果枸杞汁中可溶性固形物含量明显增加;果胶酶含量优选为0.06%,纤维素酶含量优选为0.01%,酶解最佳温度:30℃,酶解最优时间:90 min;超滤膜材料选用孔径为20 nm陶瓷膜,温度30℃～40℃,压力0.1 MPa条件时达到超滤最佳效果。反渗透温度40℃,压力4.0 MPa时达到最佳效果。超滤与反渗透浓缩连用黑果枸杞浓缩果汁除菌率可以达到5个对数值。     

加工过程对NFC苹果浊汁营养品质的影响

NFC(非浓缩还原)果汁因其丰富的营养而受到普遍关注。以国光苹果为原料,经打浆、酶解、过滤和杀菌等工艺环节制备NFC苹果浊汁,探讨主要加工环节对苹果浊汁营养品质的影响。结果表明:加工过程中苹果浊汁可溶性糖和还原糖逐渐上升;可溶性蛋白以及果胶含量逐渐下降;可溶性固形物、多酚以及黄酮含量呈先升高后下降的趋势。苹果浊汁加工过程中,多项营养指标间的变化显著相关。酶解过程对可溶性蛋白和可溶性果胶影响较大;过滤过程对蛋白、果胶、黄酮等颗粒构成物的影响较大;杀菌过程使总酚、黄酮等生物活性物质明显减少。旨在为NFC苹果浊汁加工过程中营养素的保持提供理论借鉴,为国光苹果的加工技术应用奠定理论基础。     

中空纤维膜超滤浓缩糖化酶发酵滤液

用天津纺织工学院膜分离研究所提供的聚砜中空纤维超滤膜及组体，批量浓缩糖化酶发酵滤液，酶单位浓缩倍数８．１５，浓缩体积倍数９．７３，浓缩液的酶截留率为９８．７５％，浓缩总的酶活回收率为８５．８３％。比硫酸铵盐析法工艺的收得率７２．４０％提高１３．４３％。获得了较好的效果。本文还提供了超滤浓缩糖化酶发酵滤液的技术操作要点。     

甘南牦牛肝过氧化氢酶超滤纯化工艺及酶学性质研究

采用超滤工艺从甘南牦牛肝中分离纯化过氧化氢酶,得出的超滤最佳工艺为超滤膜截留分子质量250 kDa、超滤温度25 ℃、压力0.20 MPa、流速30 m/s、浓缩倍数为3倍。酶学性质研究结果显示,该酶最适温度、pH值分别为35 ℃和7.5,米氏常数（Km）值为53.8 mmol/L。     

Changes in Apple Juice Flavor Compounds During Processing

Apple juice flavor fractions were collected at several stages during a commercial process. Replicated experiments were designed to collect samples from untreated pulp, from enzyme treated pulp, after pectinase treatment, after microfiltration (MF), and after pasteurization. Additional juice heating experiments were carried out. Six apple juice flavor compounds were identified based on to retention times and reported studies. The flavor profile changed at the different stages of the production line. Temperatures used (～57.2°C) for enzyme activation and MF tended to increase flavor compounds in apple juice except iso-butyl acetate. The higher temperatures used (～85°C) for pasteurization decreased all flavor compounds except propyl butyrate which increased. Results indicate apple juice may contain heat activated precursors of identified flavor compounds.     

Effect of processing on post-bottling haze formation in apple juice

Post-bottling haze formation in clarified apple juice produced under a variety of processing conditions was studied. Turbidity of the juice was monitored at intervals during a 6-month storage period. Changing the pasteurization temperature of raw juice and omitting the addition of amyloglucosidase did not retard or prevent haze development. Omission of the fining procedure, however, resulted in an increase in turbidity of 1.36 NTU and visual observation of larger quantities of haze. Fining with a gelatin/bentonite ratio of 1:1 gave the least haze. Thermocoagulation treatment applied to the juice prior to ultrafiltration also did not retard or prevent haze formation. Ultrafiltration at lower processing temperatures resulted in the development of less haze. At higher temperatures an increase in turbidity of over 14 NTU was observed. The results suggest that optimization of the fining procedure and ultrafiltration could be used to retard post-bottling haze in apple juice.     

Electrophoretic and immunochemical determination of bioindustrial enzymes in apple juice

The present study was undertaken to develop a specific and sensitive method which allows the detection of trace amounts of the potentially allergenic bioindustrial enzymes Pectinex Ultra-SPL (pectinase) and Gamylo 200L (amylase) in apple juice. This was achieved by an immunoblotting procedure based on sodium dodecyl sulfate (SDS) electrophoretic separation, followed by an immunochemical detection step using polyclonal rabbit antisera directed against these enzymes and, secondly, alkaline phosphatase labelled antibodies in combination with a highly sensitive enhanced chemiluminescent (ECL) detection method. In a juice spiked with enzyme at various concentrations between 10,000 and 1 ppm (1.0-0.0001%), the detection limit was 10 ppm (=0.001%) for Pectinex Ultra-SPL and 1 ppm (=0.0001%) for Gamylo 200L, respectively. Since the enzyme preparations contain less than 5% of enzymes, the detection limit for the pure enzymes is markedly lower, at least below 1 ppm. Whereas in the samples withdrawn during the initial stages of the industrial production process of apple juice, Pectinex Ultra-SPL and Gamylo 200L components were detectable in concentrations between 50 and 100 ppm, after clearing, ultrafiltration and pasteurization, neither pectinase nor amylase was detectable in the final consumer's product. Under the conditions employed, the bioindustrial enzymes Pectinex Ultra-SPL and Gamylo 200L were virtually removed during the course of production. The risk of sensitizing consumers by intake of apple juice seems to be marginal. This view is supported by the results of an animal model (anaphylaxis test), in which neither gastro-enteral reactions in guinea pigs and mice, nor IgE or IgG production by orally feeding these animals with apple juice concentrate, could be induced.     

Effect of ultrafiltration on concentrated apple juice colour and turbidity

Colour and turbidity of conventional bentonite-gelatin clarified apple juice was compared with 30,000–50,000 and 100,000DA hollow fibre ultrafiltered apple juice, with and without pectinolytic enzyme treatment. Results indicated that objective quality of ultrafiltered apple juice compared favourably with conventionally clarified apple juice over 30 weeks of storage at 37.5°C. However, treatment with pectinolytic enzymes before ultrafiltration increased both the initial colour and the browning rate. No significant differences were found in the values of the transmission haze after storage.     

Application of biosensors in early detection of contamination with lactic acid bacteria during apple juice and concentrate production

The aim of Collective Research Project QUALI-JUICE (COLL-CT-2005, Co Nr 012461) was to introduce the biosensors in control of microbiologically pure apple juice production. Three commercial lactate biosensors were used and compared with enzyme kits assays. Parallel the microbiological analysis of the production process at one juice producing enterprise was done. The results of lactic acid assay with biosensors were in good correlation with those obtained by enzyme kits. The main benefit of biosensor use was shortening of measurement time as compared with assay by enzyme kit and possibility to measure at line. The concentration of l-lactic acid in apple pulp could be correlated with the number of lactic acid bacteria. Pasteurization process was eliminating lactic acid bacteria and the concentration of lactic acid was at this stage not exceeding 0.1 g L⁻¹. The final product (apple concentrate) contained in some cases very high amounts of lactic acid indicating secondary microbiological contamination after pasteurization step. Parallel microbiological analysis of production process and lactate assay indicated that the critical point during production was prolonged vacuum filtration after pasteurization.     

Comparing the impact of high pressure,pulsed electric field and thermal pasteurization on quality attributes of cloudy apple juice using targeted and untargeted analyses

The impact of low-oxygen spiral-filter press technology combined with thermal pasteurization (TP), pulsed electric field (PEF) and high pressure processing (HPP) on cloudy apple juice quality was investigated immediately after the treatments and after 3 weeks of storage at 4 °C. Based on equivalent levels of microbial safety and desired shelf-life, low and high processing intensities were selected: TP (72 °C/15 s; 85 °C/30 s), PEF (12.5 kV/cm, 76.4 kJ/L; 12.3 kV/cm, 132.5 kJ/L), and HPP (400 MPa/3 min; 600 MPa/3 min). High intensity thermal treatment resulted in a bright, yellowish color which was maintained during storage. PPO and POD activities were largely reduced by high intensity PEF and TP yet showed high resistance to HPP. The highest vitamin C content was provided by fresh juice followed by PEF-treated juices. Due to oxidative degradation reactions, vitamin C of all treated samples significantly decreased during storage. Immediately after processing, high cloud stability values were obtained in all samples; however, cloud stability decreased during storage particularly for HPP juices with high residual PME. No significant changes were observed in pH, titratable acidity, organic acid and sugar content which also corresponded to sweet and sour taste. Results from untargeted volatile profiles showed that esters increased after PEF and were better retained after HPP. Contrary to TP treatment where ester degradation reactions occurred together with the formation of off-flavors. Most of the volatiles decreased during storage which could be linked to oxidation and ester hydrolysis reactions.Industrial relevanceBeing one of the most popular fruit juices consumed worldwide, cloudy apple juice can still undergo quality changes such as color degradation, cloud loss (fast sedimentation) and flavor changes during processing and storage. This study evaluates the potential of low-oxygen spiral-filter press in combination with different preservation technologies to obtain a maximal quality of cloudy apple juice. Results showed that high intensity thermal pasteurization can effectively inactivate quality-degrading enzymes, therefore it is useful to obtain an optimal cloudy apple juice product in terms of color and cloud stability. Although HPP has minimal impact on aroma of the juice, shelf-life of the juice may be limited due to incomplete enzyme inactivation. In the case of PEF treatment, thermal effects may contribute to maintain apple juice quality.     

固定化果胶酶提高苹果出汁率的研究

采用固定化果胶酶处理红富士苹果浆,在苹果浆pH值、固定化果胶酶水解温度、固定化果胶酶使用质量和固定化果胶酶水解时间对苹果浆出汁率的影响等单因素试验的基础上,采用响应面法分析,探索固定化果胶酶提高苹果浆出汁率的最佳条件。最佳工艺条件为苹果浆适宜pH3.43、固定化果胶酶与苹果浆质量比为1:15、酶促反应温度49.4℃、酶促反应时间3.50h。固定化果胶酶反复使用10次时,苹果浆的出汁率为62.421%,与对照组相比仍提高约13%。     

巴氏杀菌对NFC苹果汁杀菌效果及品质的影响

为研究巴氏杀菌对非浓缩还原汁(NFC)苹果汁的杀菌效果及动力学规律,对不同杀菌温度时间下NFC苹果汁菌落总数变化进行测定,并通过总酸、VC、色泽等品质指标进行分析,探讨不同杀菌条件对NFC苹果汁品质的影响。结果表明,随着加热处理温度的升高以及时间的延长,杀菌效果显著增强。对不同热处理条件下的NFC苹果汁的杀菌效果进行动力学分析,均符合一级动力学模型,拟合后各决定系数R²均大于0.9。建模分析表明,在70,75,80,85和90℃下保温58.14,44.64,18.28,9.92和4.96 min,可使微生物下降一个数量级。而要达到相同致死效果,并使灭菌时间减少到原来的1/10,所需升高的温度为17.61℃。此外,对巴氏杀菌后NFC苹果汁进行品质分析,与未处理相比,其在总酸、pH、可溶性固形物方面无显著影响,高温短时杀菌能更好保持果汁的色泽和营养成分。     

苹果浓缩汁非酶褐变动力学及影响因素

通过检测苹果汁的色值(T440nm)、褐变指数(A420nnm)、氨基态氮、5- 羟甲基糠醛含量等指标研究加工过程对苹果汁褐变的影响以及在不同贮藏温度(5、20℃和37℃)下苹果浓缩汁(70.5°Brix)的非酶褐变速率。3 个贮藏温度下的苹果浓缩汁褐变速率均符合一级反应动力学模式,而且高温情况下褐变更明显。随着贮藏时间的延长,氨基态氮含量明显下降而5- 羟甲基糠醛含量上升,尤其是贮藏在37℃的苹果汁这一变化更明显。5- 羟甲基糠醛与果汁褐变效应呈指数关系,可以说明苹果汁在贮存中的褐变主因是发生美拉德反应,而且氨基态氮含量与果汁褐变效应的线性关系也验证了这一结论。在果汁的初始加工过程中,破碎、压榨、酶解、超滤、前巴氏杀菌等单元操作中美拉德反应并不明显;吸附操作可以去除果汁中的氨基态氮以及其他色素物质;而其他操作过程如浓缩和后巴氏杀菌则促进了美拉德反应,导致果汁氨基态氮含量明显下降。     

Production of Apple Juice by Single Pass Metallic Membrane Ultrafiltration

A new process utilizing single pass metallic membrane ultrafiltration (UF) was developed to produce apple juice. Pureed whole apples were treated with cellulase and pectinase at 50°C for 2 hr, resulting in a 60% decrease in viscosity. To obtain clarified juice, enzyme treated puree was pumped through a single-pass, tubular membrane system consisting of a metallic oxide membrane formed-in-place on the porous structure of sintered stainless steel tubes. Juice yields of 80–85% were obtained with a single pass, and gas chromatographic profiles of ultrafiltered juice were nearly identical to that of the original apple puree. Permeabilities and sugar rejection characteristics of the UF system were dependent on tube diameter.