水酶法提取大豆油脂的中试研究

为了提高水酶法油脂提取率,该文进行水酶法提取大豆油脂的中试和循环酶解试验,验证小试和扩大试验的酶解和破乳参数对中试油脂提取率的影响,并研究循环酶解过程中循环次数及酶添加量对油脂提取率影响和中试工艺中大豆油脂的品质。以大豆为原料,由水酶法提取大豆油的中试得出:脱皮大豆比未脱皮大豆有助于提高油脂提取率和破乳率(P0.05,油脂提取率:脱皮61.85%,未脱皮56.91%;破乳率:脱皮92.01%,未脱皮90.42%)。通过主成分分析可知:大豆油脂的质量分数、离心后游离油和乳状液的质量分数、蛋白质的质量分数和乳状液中水分都是影响油脂提取率的关键因素。采用中试工艺1(1次提取,2次离心,1次破乳)进行循环酶解,随着循环试验次数增加,油脂提取率逐渐增大,在第4次时达到最大值66.46%±0.28%。通过检测大豆油脂的特征值可知:水酶法提取的大豆油脂的酸值、过氧化值、磷含量显著低于溶剂浸提法(P0.05),说明水酶法提取的大豆油品质高于溶剂浸提法,并且除水分和挥发物质量分数外,其他指标均符合国家三级大豆油标准。研究结果为产业化推广提供了理论参考。     

Enzyme-assisted aqueous extraction of oleosomes from soybeans (Glycine max)

Oleosomes, with their unique structural proteins, the oleosins, are known to be useful in cosmetics and other emulsion applications. A procedure to fractionate intact oleosomes to produce soybean oil without the use of organic solvents was investigated. Process parameters, enzyme treatment, filtration, cell lysis, and centrifugation, were studied. Successive extractions of the residue, eliminating the filtration step, pressurization, or ultrasonication of soybean flour prior to enzyme treatment and enzyme treatment on the residue, were the key steps. A mixture of Multifect Pectinase FE, Cellulase A, and Multifect CX 13L in equal proportion gave 36.42-63.23% of the total soybean oil from oleosomes, respectively, for 45 and 180 s of blending time, compared to the conventional method with lower yields (34.24 and 28.65%, respectively, for 45 and 180 s of blending time). Three successive extractions of the residue increased the oil yield to a maximum of 84.65% of the total soybean oil recovered in intact oleosomes. The percentage of lipid in the supernatant fraction decreased to a minimum value of 0.33% with the use of the enzymes at a 3% dosage. The results are considered to be useful for developing large-scale and efficient extraction of oleosomes from soybean.     

CaCl2和pH值对水酶法提取大豆油形成乳状液破乳效果影响

为探明粗酶水相提取大豆油所产乳状液的破乳机制,通过破乳率、Zeta电位、黏度、粒径分布和平均粒径指标分别考察无机盐和pH值对乳状液稳定性的影响。为了比较无机盐的破乳效果,该文在乳状液中分别添加浓度均为0.06mol/L的CaSO_4、CaCl_2、MgCl_2、NaCl,80℃条件下反应10min,结果显示4种无机盐均可显著降低乳状液稳定性,其中CaCl2破乳率最高,然后依次为CaSO_4、MgCl_2、NaCl。尽管CaCl2在60、70、80℃时均可实现彻底破乳,但破乳率随CaCl_2浓度(0.02～0.08mol/L)、反应时间(0～90min)、反应温度(60～80℃)的增加而提升。CaCl_2实现彻底破乳后,破乳率随反应时间延长而下降。添加CaCl_2后乳状液的电位绝对值和黏度降低,油滴发生聚合,平均粒径增加,使乳状液稳定性下降。CaCl_2浓度和反应温度的提升均可导致电位绝对值和黏度下降程度增强,破乳率进一步上升。在50℃、pH为值3～9时,降低pH值可使乳状液电位绝对值和黏度显著下降,导致油滴平均粒径增加,乳状液稳定性下降。pH值为3~4时乳状液的电位绝对值最低,接近0,此时乳状液稳定性最低,破乳率最高。但当pH值小于3时,乳状液电位绝对值和黏度再次升高,致使油滴平均粒径和破乳率降低。光镜照片显示破乳后乳状液中油珠直径明显增大。该研究可为水酶法提取大豆油破乳技术提供理论依据。     

Imbibition of soybean seeds in warm water results in the release of copious amounts of Bowman-Birk protease inhibitor, a putative anticarcinogenic agent

Protease inhibitors play a protective role against pathogenic microorganisms and herbivorous insects. The two predominant protease inhibitors of soybean seeds are the Kunitz trypsin inhibitor (KTI) and Bowman-Birk protease inhibitor (BBI). In this study, we report that soybean seeds incubated in warm water release large amounts of proteins into the surrounding media. Two-dimensional gel electrophoresis analysis of the seed exudates resulted in the separation of 93 distinct protein spots out of which 90 spots were identified by LC-MS/MS. The basic 7S globulin and the BBI are the two predominant proteins found in the soybean seed exudates. In addition to 7S and 11S seed storage proteins, others known to protect the seeds against pathogens and pests including KTI, peroxidase, α-galactosidase, and endo-1.3-β-glucanase were also identified in the seed exudates. Soybean seed exudate obtained by incubating the seeds in warm water was also able to inhibit the growth of human breast cancer cell line MCF-7. Since soybean seeds release large amounts of enzymatically active BBI when immersed in warm water, our procedure could be exploited as a simplified alternative method for the preparation of BBI concentrate which is being used as a cancer chemoprotective agent.     

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on Hypothenemus hampei gut proteinases in vitro

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme activities from H. hampei larval midgut can be inactivated by proteinaceous enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk class was purified from a wild accession of Phaseolus coccineus L. seeds. The inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH. MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da) or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1 has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a promising tool to make genetically modified coffee with resistance to coffee berry borer.     

Antioxidant activity of organic extracts from aqueous infusions of sage

The antioxidant activity of aqueous infusions of sage emerges from specific components present in that herb. In an attempt to investigate the chemical nature and properties of these components, four organic solvent extracts from aqueous infusions of sage were examined. HPLC analyses of these extracts led to the separation of a number of components, of which four were identified and quantified through the use of standard compounds of known chromatographic HPLC profiles. These compounds are the diterpenes carnosic acid, carnosol, and rosmanol and the hydroxycinnamic acid caffeic acid. The antioxidant activity and polyphenol content were determined in the four organic solvent extracts and the left-over aqueous fraction. Both polyphenolic and nonpolyphenolic substances present in the extracts arise as significant contributors to the observed antioxidant activity of the derived extracts and thus sage itself. In this sense, they reflect the antioxidant potential of the aqueous infusions of sage toward reactive oxygen species generated through variable mechanisms of iron-promoted oxidative processes.     

茄子秸秆水提物自毒作用初探

试验研究茄子秸秆水提物自毒作用结果表明,茄子干秸秆水提物原液浓度为0 .0 15 g/ mL时明显抑制茄时明显抑制茄子胚根的生长,低浓度提取物原液对胚芽生长有一定促进作用,当其浓度增至0 .0 75g/ mL时明显抑制茄时茄子胚芽生长也受抑,表明茄子秸秆水提物对茄子生长有较强的自毒作用;水提物中醇溶性组分对茄子胚根、胚芽生长的影响与水提物原液表现一致,且对水培茄子幼苗生长有较强抑制作用,表明醇溶性组分中含有大量自毒物质。茄子幼苗根系活力随水提物中醇溶性组分浓度的增加而降低,植株对NO-3 、PO3 -4、K 、Mg2 和Ca2 等离子的吸收受阻,因而抑制离子的吸收是水提物中自毒物质发挥作用的重要因素之一。     

水相酶解法提取菜籽油与菜籽蛋白工艺的优化

为提高菜籽蛋白的利用率，采用水相酶解法提取菜籽油与菜籽蛋白，以超滤法分离菜籽蛋白，重点对酶处理的工艺参数进行了研究。试验对各种酶的作用效果进行了比较，确定采用纤维素酶、果胶酶按活力比3∶1配方的复合酶处理效果较好；由优化试验得出酶处理的工艺参数为：固液比1∶5，酶用量30 U/g，酶处理时间100 min；在最佳工艺条件下获得高质量的菜籽油与菜籽蛋白，油与蛋白质得率分别为92.6%、82.3%。     

Nitrogen lowers the sulfur amino acid content of soybean (Glycine max [L.] Merr.) by regulating the accumulation of Bowman-Birk protease inhibitor

Soybeans in general contain 35-40% protein. Efforts are underway to increase further this protein content, thus enhancing their nutritive value. Even though higher protein is a desirable characteristic, whether such an increase will be accompanied by enhanced protein quality is not known. Soybean protein quality could be significantly improved by increasing the concentration of the sulfur-containing amino acids, cysteine and methionine. To ascertain if a correlation existed between protein quantity and quality, a comparison of the amino acids of soybeans differing in protein content was made. Soybeans with higher protein content had a significantly lower percentage of sulfur amino acids, while those with lower protein exhibited a higher content of cysteine and methionine. Nitrogen application elevated the protein content but lowered that of the sulfur amino acids. Transmission electron microscopy examination of thin sections of low protein soybean seeds revealed several protein storage vacuoles that were partially filled with storage proteins. Fluorescence two-dimensional difference gel electrophoresis of soybean seed proteins revealed that nitrogen application favored the accumulation of the beta-subunit of beta-conglycinin while decreasing the accumulation of Bowman-Birk protease inhibitor (BBI), a protein rich in cysteine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 60% 2-propanol-extracted proteins showed a drastic reduction in the accumulation of BBI with increasing protein content. Northern blot analysis indicated that nitrogen had a negative influence on the expression of the BBI gene. Our results indicate that the negative correlation between total protein and sulfur amino acid content is mostly mediated by the differential accumulation of BBI.     

Antioxidant properties of aqueous extracts from selected lamiaceae species grown in Turkey

Water-soluble extracts from black thyme (Thymbra spicata L.), savory (Satureja cuneifolia Ten.), Spanish oregano (Coridothymus capitatus (L.) Reichb. f.), sweet marjoram (Majorana hortensis Moench), Syrian oregano (Origanum syriacum L.), Toka oregano (Origanum minutiflorum O. Schwarz et P. H. Davis), and Turkish oregano (Origanum onites L.) were screened for antioxidant properties in a battery of six in vitro assays. Total phenol content and qualitative-quantitative compositional analyses were also carried out. The extracts demonstrated varying degrees of efficacy in each screen. The savory extract was the most effective at reducing iron(III), scavenging 1,1-diphenyl-2-picrylhydrazyl radicals, inhibiting ascorbate-iron(III)-catalyzed hydroxyl radical-mediated brain phospholipid peroxidation, and site-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. The Syrian oregano extract was the most effective chelator of iron(II), while Spanish and Turkish oregano extracts were the most effective inhibitors of nonsite-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. All the extracts contained Folin-Ciocalteu reagent-reactive substances, which was confirmed by the presence of polar phenolic analytes (i.e., hydroxybenzoates, hydroxycinnamates, and flavonoids).     

不同种植模式土壤水浸提液对胡麻的化感效应

采用浸提液生物测试法研究了不同种植模式土壤水浸提液对胡麻的化感效应。结果表明:1)不同处理土壤水浸提液对胡麻种子萌发有不同程度的抑制作用,高浓度处理比低浓度处理抑制作用更强,且抑制效应由大到小依次是胡麻连作胡-麦间作撂荒小麦连作,说明胡麻连作更能产生连作障碍,自毒效应明显;2)不同处理土壤水浸提液对胡麻幼苗的苗鲜重、根鲜重、根长有不同程度的抑制作用,且高浓度主要抑制苗鲜重、根鲜重和根长,低浓度主要抑制根鲜重、根长,胡麻连作和撂荒处理对幼苗生长抑制要大于胡-麦间作和小麦连作处理;3)不同处理土壤水浸提液对胡麻影响的综合效应随浸提液浓度的升高而增大,而小麦连作高浓度处理比低浓度处理抑制作用变化不明显,且综合效应由大到小依次是胡麻连作撂荒胡-麦间作小麦连作;4)不同种植模式处理土壤可以减轻胡麻连作产生的化感效应,处理顺序为小麦连作胡-麦间作胡麻连作。综上所述,胡麻连作障碍问题确实存在,合理的作物布局有利于改善由于胡麻自身化感物质积累所造成的连作障碍。     

Effects of solvent polarity and acidity on the extraction efficiency of isoflavones from soybeans (Glycine max)

Soy isoflavones have been correlated with beneficial health effects. The predominant chemical forms of isoflavones present may affect their biological activities. Choosing the solvent system that can accurately quantify the amounts of individual isoflavones present in these products is paramount. Our objectives were to compare frequently used solvent systems and to evaluate the effects of polarity and acidity on the recovery of isoflavones from soybeans. Isoflavones were extracted from pulverized Manokin soybeans using six solvent systems, which are the combinations of three polarity levels (83% acetonitrile, 80% methanol, and 58% acetonitrile) and two acidity levels (nonacidified and acidified). The pulverized soybean was stirred for 2 h in each solvent system before filtration and concentration using rotary evaporation. The extract was resuspended in 16% acetonitrile and analyzed by high-performance liquid chromatography. Recoveries of pure standards were evaluated with all solvent systems. Solvents with a higher polarity extracted a significantly higher amount of total isoflavones. For individual isoflavones, 58% acetonitrile (highest polarity) extracted either the highest amounts or no less than other solvents, while 83% acetonitrile (lowest polarity) extracted either the lowest amounts or no more than other solvents except for the aglycone form. Acidification significantly reduced the recovery of the malonylglucoside form and the total isoflavones. The recovery study revealed that acidification favored the chemical transformations of isoflavones during the extraction. Among the six solvent systems examined, 58% acetonitrile aqueous solution without acid was the best for extraction of isoflavones from soybeans.     

Protease extraction from soil by sodium pyrophosphate and chemical characterization of the extracts

Two arable soils and one pasture soil had previously been air-dried for 6 d and stored at room temperature. The enzyme activities remaining after this treatment were constant. The soils were then extracted with 140 mM sodium pyrophosphate at pH 7.1. Amino acid N and total organic C content of soils and soil extracts, together with humic and fulvic acids content of soil extracts were determined. Total organic C was determined in soil residues obtained after extraction. Chemical characterization of the organic matter of soils, soil extracts and soil residues was carried out by pyrolysis–gas chromatography (Py–GC). Protease activity was determined in soil extracts and soil residues by using three different substrates: N-benzoyl- -argininamide (BAA), specific for trypsin; N-benzyloxycarbonyl- -phenylalanyl -leucine (ZPL), specific for carboxypeptidases, and casein, essentially non-specific. Comparative studies between specific activities referred to organic C in soils, soil extracts and soil residues and their corresponding pyrogram composition, and also between total extracted or residual activity and the humine or unhumified organic matter content of the corresponding soil, allowed us to establish hypotheses about the type of organic matter the enzymes are associated with. From 12% to 21% of the soil organic C (33% to 39% of which were humic acids) and from 3% and 18% of amino acid N were extracted from soil using pyrophosphate. Py–GC analyses showed that pyrophosphate was effective in extracting condensed humic substances and glycoproteins and that the organic matter present in soil extracts was especially rich in intact or partially-decomposed fresh residues of carbohydrate origin and also in certain humus-associated proteins. Extracted BAA-hydrolysing activity accounted for 11% to 36% of the soil activity, depending on soil type. Extracted ZPL- and casein-hydrolysing activities were, with one exception, remarkably high, accounting for about 100% or even more of the soil activity, depending on soil type. According to the results BAA-hydrolysing proteases are probably mostly associated with highly condensed humus, ZPL-hydrolysing proteases with less condensed humic substances and casein-hydrolysing proteases with fresh organic matter.     

Purification and identification of a protease inhibitor from glassfish (Liparis tanakai) eggs

Two protease inhibitors of 67 and 18 kDa, respectively, were purified from glassfish, Liparis tanakai, eggs by affinity chromatography. The smaller protein was purified with a yield and purity of 0.25% and 49.69-fold, respectively, and was characterized for further study. The glassfish egg protease inhibitor exhibited stability between 50 and 65 degrees C in an alkaline environment (pH 8). It was shown to be a noncompetitive inhibitor against papain, with an inhibitor constant (Ki) of 4.44 nM. Potent glassfish protease inhibitor with N-Val-Gly Ser-Met-Thr-Gly-Gly-Phe-Thr-Asp-C amino acid residues was synthesized and its inhibitory activity was compared. Moreover, the 18-kDa protein inhibited cathepsin, a cysteine protease, more effectively than did egg white protease inhibitor, whereas the reverse was true for papain. Glassfish egg protease inhibitor is classified as a member of the family I cystatins.     

Phosphorus concentrations in sodium bicarbonate and citric acid extracts from arid sandy soils

Abstract

The aim of this study was to compare 1% citric acid and 1% bicarbonate solutions as extracts for the determination of phosphorus in sandy arid‐zone soils, using 64 samples from the Kalahari dunefield. For 56 of these samples, measured phosphorus concentrations were <8 ppm in both extracts, and for these 56 samples the acid‐extractable and bicarbonate‐extractable phosphorus concentrations were statistically equivalent. For the remaining 8 samples, with phosphorus concentrations >8 ppm in at least one of the extracts, phosphorus concentrations in citric acid extracts were consistently greater than those in bicarbonate extracts.     

Chemical composition and protein quality comparisons of soybeans and soybean meals from five leading soybean-producing countries

Soybeans (SBs) were obtained from five leading SB-producing countries (Argentina, Brazil, China, India, and the United States), imported to the United States, and processed into soybean meal (SBM) under uniform conditions in the United States. SBs from China had the highest crude protein (CP) content while SBs and the resultant SBM from Argentina had the lowest. Additional differences in the quality of the SB and resultant SBM samples collected were noted. An additional set of SBM produced in these five countries and subjectively evaluated to be of low, intermediate, and high quality also were obtained and evaluated. Overall, SBM quality affected amino acid and mineral concentrations with differences existing both among and within countries. SBM produced in the United States had a higher CP content than SBM produced in other countries. Amino acid concentrations generally increased, and antinutritional factors decreased with increasing subjective quality assessment.     

Positional effect on protein and oil content and composition of soybeans

Soybean (Glycine max [L.] Merr.) protein and oil qualities, with respect to monogastric nutrition, have been linked to the relative abundance of specific protein subunits and fatty acids, respectively. An analysis of field-grown soybean seeds by near-infrared spectroscopy revealed significant differences in their protein and oil contents as a function of nodal position. Seed proteins from the plant apex were high in protein and low in oil content, while those from the basal region exhibited an opposite pattern of accumulation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total seed proteins revealed that the beta-subunit of beta-conglycinin content was 4-fold higher in seeds from the apical nodes than in seeds from basal nodes. The glycinin A3 polypeptide content gradually increased in successively lower nodes from the top of the plant. Its accumulation was drastically reduced when nitrogen was applied at specific growth stages. Exogenous nitrogen did not alter the pattern of beta-subunit accumulation, but accrual of the acidic and basic polypeptides of glycinin was diminished. The remaining seed storage protein components were not influenced by nodal position or nitrogen application. Gas chromatographic analysis of fatty acids indicated that only oleic (18:0) and linoleic (18:2) acids showed variability in accumulation at different nodes. Neither the abundance nor the distribution of the fatty acids was altered by nitrogen application.     

化感水稻苗期不同器官水浸提液及根系分泌物对稗草的化感作用

在自然种植条件下研究了强化感水稻品种“P1312777”不同叶期(3～6叶)、不同器官(根、茎、叶)水浸提液对稗草根长的抑制率,比较了不同极性树脂处理后水稻根系分泌物水溶液的抑制率变化。结果表明:在3～6叶期相同水稻器官比较,均以5叶期浸提液抑制率最高;5叶期水稻不同器官比较,以叶的浸提液抑制率最高;土培至6叶期的水稻根系分泌物水溶液的抑制率为47．06％,经极性树脂ADS-21处理后抑制率增强为64．71％,经非极性树脂ADS-5处理后抑制率减弱为26．47％;水培至6叶期的水稻根系分泌物水溶液的抑制率为38．24％,经ADS- 21处理后抑制率增强为58．82％,经ADS-5处理后抑制率减弱为20．59％。因此水稻“P1312777”根系分泌物中化感物质以非(弱)极性物质为主,极性物质减弱了非(弱)极性物质的化感抑草作用。     

超声辅助双水相集成提取芒果核多酚工艺优化（英）

为了获得超声辅助双水相集成提取芒果核多酚的最佳条件,该文提出一种超声提取与丙醇-硫酸铵双水相分离体系集成提取芒果核多酚的新方法,利用5水平4因素的星点设计响应面方法研究提取条件对芒果核总多酚提取率的影响。当超声功率为400W、初始温度为45℃,获得的最佳条件是59.7%丙醇浓度,23.4%硫酸铵用量,27.3min超声提取时间和23.0mL/g的液固比,在该条件下,芒果核多酚提取率为6.67%,试验值与最佳预测值(6.82%)相符,超声辅助双水相集成法获得的提取物多酚含量高于常规超声提取法。试验结果说明利用超声辅助双水相集成的方法改善多酚提取是可行性。该研究可为芒果核的利用和植物多酚的提取技术提供参考。