杏仁豆腐系列产品的研制

通过五因素三水平正交试验对不同凝固剂及其它有关凝固条件进行研究表明，杏仁与大豆浆混合可以凝固成豆腐，并确定出了合适的工艺参数，尔后对杏仁豆腐系列制品进行了工艺研究和品评试验，分别制做了混合杏仁豆腐，蔬菜汁混合杏仁豆腐和杏仁豆腐脆片。     

Thermostable and alkaline-tolerant cellulase-free xylanase produced by thermotolerant Streptomyces sp. Ab106

Cellulase-free xylanase was produced by Streptomyces sp. Ab106 using cane bagasse as the substrate at 55 degrees C. Its maximum activity was 13 IU without cellulase and mannanase activities. Its profiles were investigated. Its optimum temperature and pH were 60 degrees C and 6.0, respectively. More than 70% of its activity was remained at 60 degrees C at pH 9. This enzyme was quite stable and exhibited an active of more than 70% for 144 h at 60 degrees C, and of more than 80% for 144 h at 40 degrees C, pH 9. This thermo-tolerant and alkaline-tolerant xylanase can be used in the pulp bleaching process.     

Detoxification of jack beans (Canavalia ensiformis): I.—Extrusion and canavanine elimination

In order to remove canavanine from jack bean (Canavalia ensiformis) the authors conducted extrusion and extraction experiments. In the extrusion experiments, using jack bean without testa and conditioned to 240 g kg^?1 moisture content, different temperatures of the compression section of the extruder and screw rates were tried. Under these conditions the parameters measured were expansion, final moisture, haemagglutinins, urease activity, available lysine and soluble nitrogen index. The screw rate did not affect the extrusion of jack bean. The best temperature was 110°C. The extraction experiments were carried out with extruded jack bean (EJB) at 110°C and 70 rpm using one or two acid or formalin extractions. EJB amount, times and pH were evaluated with a surface response model in one acid extraction experiment. The best results were observed with 100 g kg^?1 of EJB, at pH 55 for 2 h (1AEJB), which reduced the canavanine content to 8.5 g kg^?1. In the same conditions using a second acid extraction (2AEJB) we obtained a product with 4.9 g kg^?1 of canavanine. With the formalin extraction at pH 5.5 and 7.5, and a molar ratio of canavanine: formalin 1 : 1 and 1 : 2, we obtained a product with 8.2 g kg^?1 of canavanine.     

Effects of food formulation and thermal processing on flavones in celery and chamomile

Flavones isolated from celery varied in their stability and susceptibility to deglycosylation during thermal processing at pH 3, 5, or 7. Apigenin 7-O-apiosylglucoside was converted to apigenin 7-O-glucoside when heated at pH 3 and 100 °C. Apigenin 7-O-glucoside showed little conversion or degradation at any pH after 5 h at 100 °C. Apigenin, luteolin, and chrysoeriol were most stable at pH 3 but progressively degraded at pH 5 or 7. Chamomile and celery were used to test the effects of glycosidase-rich foods and thermal processing on the stability of flavone glycosides. Apigenin 7-O-glucoside in chamomile extract was readily converted to apigenin aglycone after combination with almond, flax seed, or chickpea flour. Apigenin 7-O-apiosylglucoside in celery leaves was resistant to conversion by β-glucosidase-rich ingredients, but was converted to apigenin 7-O-glucoside at pH 2.7 when processed at 100 °C for 90 min and could then be further deglycosylated when mixed with almond or flax seed. Thus, combinations of acid hydrolysis and glycosidase enzymes in almond and flax seed were most effective for developing a flavone-rich, high aglycone food ingredient from celery.     

Purification and partial characterization of psychrotrophic Serratia marcescens lipase

Serratia marcescens isolated from raw milk was found to produce extracellular lipase. The growth of this organism could contribute to flavor defects in milk and dairy products. Serratia marcescens was streaked onto spirit blue agar medium, and lipolytic activity was detected after 6 h at 30 degrees C and after 12 h at 6 degrees C. The extracellular crude lipase was collected after inoculation of the organism into nutrient broth and then into skim milk. The crude lipase was purified to homogeneity by ion-exchange chromatography and gel filtration. The purified lipase had a final recovered activity of 45.42%. Its molecular mass was estimated by SDS-PAGE assay to be 52 kDa. The purified lipase was characterized; the optimum pH was likely between 8 and 9 and showed about 70% of its activity at pH 6.6. The enzyme was very stable at pH 8 and lost about 30% of its activity after holding for 24 h at 4 degrees C in buffer of pH 6.6. The optimum temperature was observed at 37 degrees C and exhibited high activity at 5 degrees C. The thermal inactivation of S. marcescens lipase was more obvious at 80 degrees C; it retained about 15% of its original activity at 80 degrees C and was completely inactivated after heating at 90 degrees C for 5 min. Under optimum conditions, activity of the enzyme was maximum after 6 min. The Michaelis-Menten constant was 1.35 mM on tributyrin. The enzyme was inhibited by a concentration more than 6.25mM. Purified lipase was not as heat-stable as other lipases from psychrotrophs, but it retained high activity at 5 degrees C. At pH 6.6, the pH of milk, purified lipase showed some activity and stability. Also, the organism demonstrated lipolytic activity at 6 degrees C after 12 h. Therefore, S. marcescens and its lipase were considered to cause flavor impairment during cold storage of milk and dairy products.     

Survival of Salmonella enteritidis phage type 30 on inoculated almonds stored at -20, 4, 23, and 35 degrees C

To evaluate the survival of Salmonella on raw almond surfaces, whole almond kernels were inoculated with Salmonella Enteritidis phage type (PT) 30 collected from a 24-h broth culture or by scraping cells from an agar lawn. Kernels inoculated with lawn-collected cells to 8, 5, 3, and 1 log CFU per almond after a 24-h drying period were stored for 161 days at 23 +/- 3 degrees C. Calculated rates of reduction were similar for the four inoculum levels (0.22, 0.28, 0.29, and 0.22 log CFU/month, respectively). Kernels inoculated to 7.1 or 8.0 log CFU per almond after drying were stored for 171 or 550 days, respectively, at selected temperatures, including -20 +/- 2 degrees C, 4 +/- 2 degrees C, 23 +/- 3 degrees C, and 35 +/- 2 degrees C. No significant reductions of Salmonella were observed during storage at -20 and 4 degrees C over 550 days. At 35 degrees C, a biphasic survival curve was observed, with calculated reductions of 1.1 log CFU/month from days 0 to 59 and no significant reduction from days 59 to 171. At 23 degrees C, reductions of 0.18 and 0.30 log CFU/month were calculated for 171 and 550 days of storage, respectively. When combined with data from the study of inoculum levels, an overall average calculated reduction at 23 degrees C was 0.25 +/- 0.05 log CFU/month. Significantly greater reductions were observed during the 24-h drying period when broth-collected cells were used as the inoculum, suggesting that cells collected from agar lawns were more resistant to drying. However, after initial drying, the rates of reduction at 23 degrees C did not differ significantly between the inoculum preparation methods. Salmonella Enteritidis PT 30 survives for long periods on almond kernels under a variety of commonstorage conditions.     

Purification and characterization of the lipase from Pseudomonas fluorescens HU380

A lipase, which markedly splits polyunsaturated fatty acid ester (PUFA) bonds, from newly isolated Pseudomonas fluorescens HU380 was purified. The purification procedure included Phenyl-Toyopearl fractionation, DEAE-Sepharose chromatography, and Superdex-200HR chromatography. The enzyme was purified 24.3-fold with a yield of 14% and a specific activity of 9854 U/mg. Its molecular weight was estimated on SDS-PAGE to be 64,000. The optimum pH and temperature were 8.5 and 45 degrees C, respectively. The lipase was stable over the pH range of 6.0-7.0 at 30 degrees C for 24 h, and up to 40 degrees C at pH 7.0 for 60 min, when 0.1% Triton X-100 was present. The lipase preferably acted on short to middle-chain fatty acid simple methyl-esters and triglycerides, and cleaved mainly 1,3-ester bonds and to a lesser extent the 2-position ester bond of triolein. The lipase was inhibited by Co2+, Ni2+, Fe3+, Fe2+, and EDTA, and activated by Ca2+. Its N-terminal amino acid sequence was determined to be GVYDYKNFGTADSKALFSDAMAITLY, which exhibited considerable similarity with those of the lipases from other P. fluorescens strains, but no significant homology with other lipases. This lipase was able to decompose fats and oils that contained eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) without significantly affecting the contents of these fatty acids. The results suggest that the lipase may be useful when applied to the processing of industrial fats and oils containing EPA and DHA, such as fish oil splitting.     

Preparation of lactose‐free milk by using salt‐fractionated almond (Amygdalus communis) β‐galactosidase

β‐galactosidase was isolated from almond (Amygdalus communis) extract by ammonium sulfate precipitation. Almond proteins precipitated by using ammonium sulfate and then dialysed exhibited 5.3‐fold purification of β‐galactosidase, and the yield of enzyme preparation was 96.5%. The partially purified β‐galactosidase exhibited pH and temperature optima at pH 5.5 and 50 °C, respectively. The enzyme was significantly stable against heat, pH, calcium and magnesium ions and D ‐galactose. The almond β‐galactosidase preparation exhibited over 89% activity even after 2 months storage at 4 °C. Hydrolysis of lactose in milk and whey was performed in a stirred batch process by using this enzyme preparation. These observations indicated that the hydrolysis of lactose increased continuously with time. The enzyme could hydrolyse 94% of lactose in buffer solution and whey whereas 90% of lactose hydrolysis was achieved in milk. The main aim of the present study was to prepare lactose‐free milk, which must be free from contamination, and the process should be inexpensive. Copyright © 2007 Society of Chemical Industry     

Purification and properties of hydrogen sulfide oxidase from Bacillus sp. BN53-1

A hydrogen sulfide oxidase was purified to homogeneity from the heterotroph Bacillus sp. BN53-1 isolated from pig feces compost. The enzyme was found to be a monomer with a M(r) value of approximately 37 kDa. It required FAD for its activity, which was not replaced by FMN. The optimum reaction pH and temperature were 7.5 and 40 degrees C, respectively. The enzyme was stable between pH 6.0 and 7.0 and up to 30 degrees C. Its activity was stimulated by Ca2+ and Mn2+ and inhibited by Al3+, dithiothreitol, and 2-mercaptoethanol. The main product was elemental sulfur, and H2O2 was not detected. The N-terminal sequence of the enzyme showed similarity to other FAD-requiring enzymes.     

α-D-半乳糖苷酶源的选择及其性质的探讨

本文就植物种子中α-D-半乳糖苷酶源的选择及其性质做了较详细的探讨。作者认为在相同条件下,苦杏仁、小豆和芸豆比较,小豆中的α—D—半乳糖苷酶活力最强。小豆中含有两种α—D—半乳糖苷酶(酶Ⅰ和酶Ⅱ)。本文以对—硝基苯,α—D半乳糖酶(PNPG)为底物时其最适温度和pH分别为45～50℃5.2;45℃、5.2。酶Ⅰ的V_(m a x)和Km分别为8.33μg/ml。mln和1.25×10~(-3)M,还测得元素对α—D—半乳糖苷酶有强烈的激活作用。半乳糖是酶Ⅰ的强烈竞争性抑制剂。     

扁杏仁乳饮料的稳定性分析与工艺研究

以扁杏仁与牛乳为原料研制保健乳饮料,采用刺槐豆胶、海藻酸钠、卡拉胶、黄原胶、BE-3等稳定剂,然后对成品进行稳定性分析。结果表明,BE-3复合稳定剂的效果明显优于其他4种稳定剂。选取杏仁粉、蔗糖、BE-3稳定剂3个因素,利用L9(34)正交设计进行工艺优化。以色泽、气味、滋味、组织状态等指标进行感官评定。扁杏仁乳饮料的最佳配方为:杏仁粉添加量为4%,蔗糖添加量为7%,BE-3型稳定剂的添加量为0.45%。扁杏仁乳饮料营养丰富,纯香柔滑,是一种复合保健乳饮料。     

Purification and characterization of a monoacylglycerol lipase from Pseudomonas sp. LP7315

A monoacylglycerol lipase (MGL) was purified from Pseudomonas sp. LP7315 by ammonium sulfate precipitation, anion-exchange chromatography, and preparative electrophoresis. The purified enzyme was homogeneous on SDS-PAGE with a molecular mass of 59 kDa. Its hydrolytic activity was confirmed to be specific for monoglycerides: the enzyme did not hydrolyze di- and triglycerides. MGL was found to be stable even after 1-h incubation at 65 degrees C. The optimum pH for monopalmitin hydrolysis was approximately 8. The hydrolytic activity depended not only on temperature and pH but also on the type of monoglyceride used. MGL also catalyzed monoglyceride synthesis at 65 degrees C in a solvent-free two-phase system, in which fatty acid droplets were dispersed in the glycerol phase with a low water content. The synthetic reaction proceeded at a constant rate for approximately 24 h and approximately reached an equilibrium after 48 h of reaction. The initial rate and equilibrium yield of the synthetic reaction depended on the type of fatty acid used as the substrate.     

The influence of salt and temperature on the natural fermentation of African locust bean

Variations in salt (NaCl) content and temperature influenced the microbial development and organoleptic quality of fermenting African locust bean (Parkia filicoidea Welw.) seeds. The influences on microbial growth by different salt contents and temperatures were followed by changes of pH and titratable acidity. A 1% (w/w) salt addition and fermentation at 37 degrees C improved the organoleptic quality of the product. Salt additions above 3% (w/w) and temperatures below 25 degrees C resulted in lower microbial counts, low pH and titratable acid values; fermentation of the processed substrate of African locust bean seeds was inhibited and the organoleptic quality was poor. The predominant microorganism present throughout the fermentation was a Bacillus sp. with characteristics similar to Bacillus subtilis.     

Purification and characterization of organic solvent-stable lipase from organic solvent-tolerant Pseudomonas aeruginosa LST-03

An organic solvent-stable lipase (LST-03 lipase) secreted into the culture broth of the organic solvent-tolerant Pseudomonas aeruginosa LST-03 was purified by ion-exchange and hydrophobic interaction chromatography in the presence of 2-propanol. The purified enzyme was homogeneous as determined by SDS-PAGE. The molecular mass of the lipase was estimated to be 27.1 kDa by SDS-PAGE and 36 kDa by gel filtration. The optimum pH and temperature were 6.0 and 37 degrees C. LST-03 lipase was stable at pH 5-8 and below 40 degrees C. Its hydrolytic activity was highest against tricaproin (C6), methyl octanoate (C8), and coconut oil respectively among the triacylglycerols, fatty acid methyl esters, and natural oils investigated. The enzyme cleaved not only the 1,3-positioned ester bonds, but also the 2-positioned ester bond of triolein. It exhibited high levels of activity in the presence of n-decane, n-octane, DMSO, and DMF as well as in the absence of an organic solvent. In addition, LST-03 lipase was stabler in the presence of n-decane, ethyleneglycol, DMSO, n-octane, n-heptane, isooctane, and cyclohexane than in the absence of an organic solvent.     

Characteristics and Concentrations of α-Amylase Inhibitor in Phaseolus vulgaris Biotypes

Flours from six legume species (lentil, lima bean, field pea, chickpea, fababean, mung bean) were devoid of α-amylase inhibitory activity but five biotypes of Phaseolus vulgaris showed activities of 27 to 40 units/mg protein. Air classification of the pin milled flours yielded fine fractions which contained 37% (black bean) to 61% (navy bean) protein and 38 to 66 units of inhibitor activity/mg protein, respectively. The coarse starch fractions were proportionately depleted in protein content and inhibitor activity. The partially purified amylase inhibitor from navy bean was active towards porcine pancreatic α amylase. The optimum pH for inhibition was 5.6 - 5.7, with the inhibitor being most stable at pH 6.9 after 4 h incubation at 37°C. The inhibition was faster at 37°C than at 25°C. Although retaining most of its inhibitory power between 37°C–60°C, the navy bean inhibitor showed a complete loss of activity after 20min at 90°C.     

Modelling growth and bacteriocin production by Lactobacillus curvatus LTH 1174 in response to temperature and pH values used for European sausage fermentation processes

Lactobacillus curvatus LTH 1174, a strain isolated from fermented sausage, produces the antilisterial bacteriocin curvacin A. Its biokinetics of cell growth and bacteriocin production as a function of temperature (20-38 degrees C) and pH (4.8-7.0) were investigated in vitro during laboratory fermentations using de Man, Rogosa and Sharpe (MRS) medium. A predictive, successfully validated model was set up to describe the influence of temperature and pH on the microbial behavior. Both cell growth and bacteriocin activity were influenced by changes in temperature and pH. The optimum temperature value for cell growth, 34.5 degrees C, did not correspond with the optimum temperature for curvacin A production (20-27 degrees C). Interestingly, the pH range for growth and curvacin A production was broad. Thus, Lb. curvatus LTH 1174 seems to be a promising bacteriocin-producing strain for use in European sausage fermentations that are performed at temperatures near 25 degrees C.     

Growth of Salmonella Enteritidis phage type 30 in almond hull and shell slurries and survival in drying almond hulls

Traceback investigation of a 2000 to 2001 outbreak of salmonellosis associated with consumption of raw almonds led to isolation of the outbreak strain Salmonella enterica serovar Enteritidis phage type (PT) 30 on three geographically linked almond farms. Interviews with these growers revealed that significant rain fell during the 2000 harvest when many almonds were drying on the ground. The objectives of this study were to document weather conditions during the 2000 harvest, determine the potential for growth of Salmonella Enteritidis PT 30 in hull or shell slurries, and evaluate survival of Salmonella Enteritidis PT 30 on wet almond hulls during drying. Dry almond hulls and in-shell kernels wetted for 24 h increased in weight by 250 to 300% and 100%, respectively. Both hull and shell slurries supported rapid growth of Salmonella Enteritidis PT 30 at 24 degrees C; slurries containing hulls also supported growth at 15 degrees C. Maximum Salmonella Enteritidis PT 30 concentrations of 6.2 and 7.8 log CFU/ml were observed at 15 and 24 degrees C, respectively. Salmonella Enteritidis PT 30 grown in wet hulls that were incubated at 24 degrees C survived drying at either 15 or 37 degrees C. Reductions of 1 to 3 log CFU/g of dry hull were observed during drying; reductions generally declined as incubation time increased from 2 to 7 days. Evaluation of shipping records revealed that approximately 60% of outbreak-associated almonds had not been exposed to rain, eliminating this factor as the sole cause of the outbreak. However, the data provide evidence that wet almonds may be a greater risk for high concentrations of Salmonella, and specific guidelines should be established for harvesting and processing almonds that have been exposed to rain or other water sources.     

Purification and characterization of sea squirt alpha-N-acetylgalactosaminidase

Sea squirt alpha-N-acetylgalactosaminidase was purified to homogeneity. Its molecular weight was estimated to be approximately 160,000 by gel filtration and 40,000 by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing condition. The chromatographic and electrophoretic behaviors indicated that the enzyme was composed of four subunits. The optimum pH of the enzyme reaction was about 4.0 at 37 degrees C, while the enzyme was stable in the range of pH 5.0 to 6.0 during 4 h preincubation at 37 degrees C. Although the enzyme (0.1 unit) was stable at 0 degrees C for 30 min in the presence of 7.5 mM metal ions (Al3+, Ba2+, Ca2+, K+, Mn2+, Pb2+, Sr2+, and Zn2+), almost 40% of the enzyme activity was lost in the presence of Cu2+, Hg2+, monoiodoacetic acid, and EDTA. The enzyme hydrolyzed aryl N-acetyl-alpha-D-galactosaminide as well as GalNAcalpha1(-->4GalNAcalpha1-->)n 4GalNAc-p-aminobenzoic acid ethyl ester (ABEE) (n = 1-4), but GalNAcalpha1-->4GalNAc-ABEE only scarcely. Furthermore, an allergenic pentasaccharitol ABEE derivative, GalNAcalpha1-->2Fucalpha1-->3(GalNAcbeta1-->4) GlcNAcbeta1-->2(3-acetoamido-3-deoxy)L-threose-ABEE, the minimum structural unit for the sea squirt allergenicity was hydrolyzed to 95 mol% for 72 h incubation with the enzyme. The enzyme could be utilized as a powerful tool for the structural analyses of the carbohydrate epitopes of the sea squirt allergen molecules.     

Migration of Salmonella Enteritidis phage type 30 through almond hulls and shells

The ability of Salmonella to migrate from an external aqueous environment through the almond hull and shell, and to colonize the kernel, was evaluated in two ways. First, the outer surface of shell halves from five varieties of almonds that differed in shell hardness were placed in contact with a suspension of Salmonella enterica serovar Enteritidis phage type 30 for 24 h at 24 degrees C. Salmonella Enteritidis was isolated from the inside of these almond shells in 46 and 100% of the samples, by direct swabbing of the inner surface of the shell and by enrichment from the swab, respectively. These findings suggested that hardness of the shell is not a significant factor in the migration of the pathogen through that tissue. In addition, both motile and nonmotile strains of S. enterica serovar Typhimurium migrated through the almond shells to the same extent under the conditions of this assay, indicating that bacterial migration through the wet shell may be a passive process. Second, whole almonds (intact hull, shell, and kernel) were soaked for 24 to 72 h at 24 degrees C in a suspension of Salmonella Enteritidis phage type 30 labeled with the green fluorescent protein. Green fluorescent protein-labeled Salmonella cells were observed on the outer and inner surfaces of both the almond hull and shell, and on the kernel, by confocal laser scanning microscopy. Our data provide direct evidence that wet conditions allow for Salmonella migration through the hull and shell and onto the almond kernel, thus providing a means by which almond kernels may become contaminated in the field.     

杏仁蛋白的两级泡沫分离工艺优化

为了研究泡沫分离法提取杏仁蛋白的最佳工艺,在进料pH、进料浓度、鼓泡气速和鼓泡时间对杏仁蛋白分离效果影响的基础上,利用田口实验设计法建立了杏仁蛋白的两级泡沫分离工艺。结果表明,泡沫分离最佳工艺条件为:pH4.0、进料浓度6 g/L、气速400 mL/min,鼓泡时间10 min。在该工艺条件下,一级泡沫分离所得杏仁蛋白的质量分数为75.23%,蛋白回收率为79.48%;二级泡沫分离所得杏仁蛋白的质量分数为84.71%,蛋白回收率为71.19%。实验结果表明,两级泡沫分离法可以作为分离杏仁蛋白的一种新方法。