豆腐柴叶果胶的提取工艺条件研究

本文采用正交试验的方法研究了酸解法从豆腐柴叶中提取果胶的最佳提取工艺条件：萃取温度90-95℃、溶液pH值1．5、萃取时间50-60min、料液比1：20,在此条件下豆腐柴叶果胶的提取率为14％-17％。     

豆腐柴叶提取低酯果胶的研究

豆腐柴叶含有丰富的果胶。文中结合盐酸浸提法的基础上,采用醇氨降酯法提取豆腐柴叶的果胶并检测,提取率为部颁19.5%,pH值为2.9,水分和灰分分别为5.13%和8.67%,半乳糖含量为65%,酯化度为38.5%,所提取的低酯果胶的基本性质与国标检测标准接近。果胶成品经AB-8树脂柱纯化后,所提果胶的成分经硅胶板薄层层析初步测定,果胶多糖成分为葡萄糖,果糖,D-甘露糖3种,果胶的溶解度与温度呈正相关性,pH值对其溶解度影响不大。     

豆腐木叶果胶含量的动态变化规律研究

测定了不同季节豆腐柴木叶中的果胶含量,并研究了不同叶位豆腐木果胶含量的变化规律.结果表明,豆腐木叶中的果胶含量在6～8月呈上升趋势,8月最高,9月开始下降.不同叶位果胶含量中,第3、5、6叶位的豆腐木叶片果胶含量较高,第5叶最高.     

大豆豆腐产量的遗传研究

以六合小叶青×新沂小黑豆、上饶干不死×淮阴秋黑豆、六合小叶青×南农７３－９３５ ３个杂交组合植株世代的Ｐ１、Ｐ２、Ｆ１、Ｆ２、Ｆ２：３为材料,分析了干豆腐产量的遗传规律。结果表明,３个杂交组合干豆腐产量的遗传均是１对加显性主基因和多基因混合遗传模型,干豆腐产量的遗传率较高,３个杂交组合植株世代Ｆ２：３家系干豆腐产量的主基因遗传率分别为５１．８０％、５９．８０％、６１．８５％,多基因遗传率分别为４８．０３％、３９．１８％、３６．１２％。进行高豆腐产量选育时应以主基因选择为主,兼顾多基因的选择。     

产果胶酶的菌种选育及发酵条件

炭黑曲霉(Aspergillus carbonarius)AS 3.396经亚硝基胍和Co60r-射线诱变,获得一株高产果胶酶突变株G5512。该菌株的发酵滤液,以高酯果胶为底物的酶活力为860u／ml;以低酯果胶为底物的酶活力为1227u／ml。产酶活力水平约为原出发菌株的2—6倍。产酶最适培养条件为：起始pH4.0—4.5,30℃,72—90小时。酶作用最适条件为：高酯果胶为底物时,pH3.5,50℃;低酯果胶为底物时,pH4.5,50℃。pH稳定范围为2．0-6．5(高酯果胶)和4.5—5.0(低酯果胶)。酶在60℃保温15分钟,高酯果胶为底物的酶剩余活力71％,低酯果胶为底物的酶活力仅剩余1％。     

以豆腐柴叶为原料提取果胶工艺研究

本文开发了一条从豆腐柴叶中提取果胶的新工艺。该工艺中,树脂吸附纯化、超滤浓缩和喷雾干燥是三个重要的单元操作。为了确定最佳提取条件和考察树脂吸附纯化和超滤浓缩两个单元操作的商业应用可行性,进行了三种不同规模的试验。结果表明,所开发的工艺在能耗和果胶质量方面明显优于传统的醇沉淀法。     

洋葱种子含水量与贮藏温度对其寿命的影响

不同含水量（MC 7．1％－1．2）的洋葱种子贮藏在35℃、室温、15℃和5℃条件下1－3年,适度超于处理能延长种子的贮藏寿命;种子的贮藏寿命与种子含水量和贮藏温度密切相关。种子贮藏的最适含水量随温度的改变而发生相应的变化,35℃时MC为3．4％;室温时为3．4％－3．5％;15℃时为4．5％－5．1％。MC≤2．2％不利于延长种子寿命。在室温自然条件下贮藏1－3年,适度超干种子（MC3．4％）内MDA和H2O2含量、O2^-产生速率和LOX活性明显地低于未超干种子（MC7．1％）和高度超干种子（MC1．2％）,而抗氧化酶AsA-POD 、CAT和SOD的活性显著地高于未超干种子（MC7．1％）和高度超干种子（MC1．2％）。据此认为对脂质过氧化的抑制作用是适度超干种子耐贮藏的生理原因之一。     

乳酸菌Enterococcuse faecalis TN-9低温蛋白酶的提纯及性质

对产自乳酸菌Enterococcuze fecalis TN-9的蛋白酶,进行了硫酸铵沉淀,DEAE—Sephadex A-25以及DEAE Cellulofine A-500离子交换层析的3步纯化和特性研究。纯化酶Native PAGE显示1条蛋白带。SDSPAGE和凝胶层析分子量分别为30ku及69ku。纯化酶最适作用温度为30℃,最适作用PH为7．5～8．0,在pH6．0～9．5和45℃以下条件下稳定,在0℃下显示了6．1％的相对活性,60℃以上热处理完全失去酶活。该酶被EDTA-2Na,Hg^2＋、Cu^2＋、Ni^2＋、Ag^2＋、Co^2＋及Pepstatin A不完全抑制。Zn^2＋对蛋白酶具有明显的激活作用。纯化酶作用于偶氮酪蛋白的Km和Vmax分别为0．098％和72mg／（h·mg）。该酶为N末端VGSEVTLKNS的明胶酶（Gelatinase）的一种,性质属于低温蛋白酶。     

一种简便的提取蚕蛹蛋白新方法

以蚕蛹为原料,采用一种简便的提取工艺提取蚕蛹蛋白质,结果表明：干蛹约占鲜蛹重的32．5％,干蛹中蛋白质含量为54．2％,成品中蚕蛹蛋白质含量为83．31％,其蚕蛹蛋白收率达干蛹的61．32％,是文献报道用减法提取收率的2．27倍,蚕蛹中蛋白质的回收率达94．33％。     

类芽胞杆菌碱性果胶裂解酶的纯化与酶学性质表征

从类芽胞杆菌Paenibacillus sp.WZ008的发酵上清液中纯化得到一个高活力碱性果胶裂解酶,经SDS-PAGE电泳估算其亚基相对分子质量为4.5×104。通过对该酶进行酶学性质研究发现：该酶能催化裂解果胶酸、低酯果胶和高酯果胶;酶催化反应最适温度范围为55～60℃,最适pH为9.6,在最适条件下以低酯果胶为底物酶的比酶活达3 021.6 U/mg;Ca2＋能增强该酶的活力,而Mn2＋,Ba2＋和EDTA强烈抑制该酶活力;当没有Ca2＋存在时,高度酯化的果胶是该酶的最适底物,在4 mmol/L Ca2＋存在时,该酶以果胶酸为底物比酶活最高（25 467 U/mg）。该酶N端序列比对分析发现与类芽胞杆菌Paenibacillus amylolyticus strain 27c64果胶裂解酶高度同源。     

Identification of enzymes that are effective for isolating protoplasts from grass leaves

Cellulase C₁, cellulase Cx, and xylanase were isolated and purified from a cellulase preparation of Trichoderma viride as enzymes effective in the isolation of protoplasts from oat leaves. Pectin lyase which is specific for methyl-galacturonide linkages was also found to be a useful enzyme for the isolation of protoplasts from the tissues. This suggested that pectic polysaccharides with a high degree of esterification may play an important role in cell walls of Gramineae. It was necessary to use the mixture of cellulase C₁, cellulase Cx, xylanase, and pectin lyase for the rapid isolation of protoplasts, while a small amount of protoplasts could be isolated from oat leaves by cellulase C₁ plus xylanase or cellulase C₁ plus pectin lyase. The mixture of four enzymes also was effective in the isolation of protoplasts from the leaves of wheat, barley, and corn.     

Cell wall cementing materials of grass leaves

Treatment of grass leaves with either a purified pectin lyase of Aspergillus japonicus or a purified xylanase of Trichoderma viride could lead to the isolation of some single leaf cells. However, a mixture of pectin lyase and xylanase brought about more rapid isolation of single cells than did either of the two enzymes alone, indicating a synergistic effect. Analysis of the components released from oat cell walls by the enzymes indicated that both homogalacturonans with a high degree of esterification and a kind of glucuronoarabinoxylan with ferulic acid ester may play a role in cell wall cementing in grass leaves.     

Xylogalacturonan exists in cell walls from various tissues of Arabidopsis thaliana

Evidence is presented for the presence of xylogalacturonan (XGA) in Arabidopsis thaliana. This evidence was obtained by extraction of pectin from the seeds, root, stem, young leaves and mature leaves of A. thaliana, followed by treatment of these pectin extracts with xylogalacturonan hydrolase (XGH). Upon enzymatic treatment, XGA oligosaccharides were primarily produced from pectin extracts obtained from the young and mature leaves and to a lesser extent from those originating from the stem of A. thaliana. The oligosaccharide GalA(3)Xyl was predominantly formed from these pectin extracts. No XGA oligosaccharides were detected in digests of pectin extracts from the seeds and roots. A low number of XGA oligosaccharides was obtained from pectins of A. thaliana. This indicates a uniform distribution of xylose in XGA from A. thaliana. The predominant production of GalA(3)Xyl, as well as the release of linear GalA oligosaccharides pointed to a lower degree of xylose substitution in XGA from A. thaliana than in XGA from apple and potato. The estimated amount of XGA accounted for approximately 2.5%, 7% and 6% (w/w) of the total carbohydrate in the pectin fraction of the stem, young leaves and mature leaves, respectively.     

Purification, characterization and cloning of antiviral/ribosome inactivating protein from Amaranthus tricolor leaves

An antiviral protein (AVP), imparting high level of resistance against sunnhemp rosette virus (SRV) was purified from the dried leaves of Amaranthus tricolor. The purified protein (AAP-27) exhibited approximately 98% inhibition of local lesion formation at a concentration range of approximately 30 microg ml(-1). The protein was found to be highly basic glycoprotein monomer (pI approximately 9.8) of Mr 27 kDa, with neutral sugar content of 4%. The purified protein exhibited N-glycosidase and RNase activities. We have also isolated full-length cDNA clone, encoding this protein designated as A. tricolor antiviral protein-1 (AAP-1). Two primers, one designed on the basis of N-terminal sequence of the purified protein and the other from the conserved active peptides of other AVPs/RIPs were used for PCR amplification of double stranded cDNA, isolated from the leaves of A. tricolor. The amplified fragment was used as a probe for library screening. The isolated full-length cDNA consisted of 1058 nucleotides with an open reading frame encoding a polypeptide of 297 amino acids. The deduced amino acid sequence of AAP-1 has a putative active domain conserved in other AVPs/RIPs and shows varying homology to the RIPs from other plant species.     

Untersuchungen Zur Energetischen Verwertung Von Rationen Mit Melasse,Getrockneten Zuckerrüben,Apfelpektin Und Luzerneblatt Durch Adulte Schweine

Abstract

Von 6 Rationen mit Melasse, getrockneten Zuckerrüben, Apfelpektin und Luzerneblatt wurde der Energieumsatz an 8 adulten Schweinen mittels Respirationsversuchstechnik gemessen. Die Verdaulichkeit der Energie der Rationen lag im Bereich von 74–88%, die partielle Verwertung der umsetzbaren Energie zwischen 66 und 78%. Aus den Versuchsergebnissen mit Apfelpektin wurde ein mittlerer Ansatzwert von 7,2 ± 12,5 kJ/g verdauliches Pektin abgeleitet. Unter Berücksichtigung der Literaturbefunde und der hohen Standardabweichungen wird für verdauliches Pektin ein energetischer Wirkungswert von 5 kJ/g für die Schätzung des energetischen Futterwertes (Nettoenergie-Fett) empfohlen.

STUDIES OF THE ENERGETIC UTILIZATION OF RATIONS WITH MOLASSES, DRIED SUGAR BEETS, APPLE PECTIN AND LUCERNE LEAVES IN ADULT PIGS

The energetic utilization of 6 rations with molasses, dried sugar beets, apple pectin and lucerne leaves was measured in 8 adult pigs by means of the respiration test technique. The energy digestibility of the rations ranged from 74 to 88%, the partial utilization of the metabolizable energy from 66 to 78%. From the results with apple pectin a mean energy deposition value of 7.2 ± 12.5 kJ/g digestible pectin was derived. Taking into consideration results from previous recorded experiments and the high standard deviation for estimation of the energetic feed value as deposition effect of digestible pectin, 5 kJ/g are proposed.     

Pectinase production by Neurospora crassa: purification and biochemical characterization of extracellular polygalacturonase activity.

The production of pectinase was studied in Neurospora crassa, using the hyperproducer mutant exo-1, which synthesized and secreted five to six times more enzyme than the wild-type. Polygalacturonase, pectin lyase and pectate lyase were induced by pectin, and this induction was glucose-repressible. Polygalacturonase was induced by galactose four times more efficiently than by pectin; in contrast the activity of lyases was not affected by galactose. The inducing effect of galactose on polygalacturonase was not glucose-repressible. Extracellular pectinases were separated by ion exchange chromatography. Pectate and pectin lyases eluted into three main fractions containing both activities; polygalacturonase eluted as a single, symmetrical peak, apparently free of other protein contaminants, and was purified 56-fold. The purified polygalacturonase was a monomeric glycoprotein (38% carbohydrate content) of apparent molecular mass 36.6-37.0 kDa (Sephadex G-100 and urea-SDS-PAGE, respectively). The enzyme hydrolysed predominantly polypectate. Pectin was also hydrolysed, but at 7% of the rate for polypectate. Km and Vmax for polypectate hydrolysis were 5.0 mg ml-1 and 357 mumol min-1 (mg protein)-1, respectively. Temperature and pH optima were 45 degrees C and 6.0, respectively. The purified polygalacturonase reduced the viscosity of a sodium polypectate solution by 50% with an increase of 7% in reducing sugar groups. The products of hydrolysis at initial reaction times consisted of oligogalacturonates without detectable monomer. Thus, the purified Neurospora crassa enzyme was classified as an endopolygalacturonase [poly(1,4-alpha-D-galacturonide) glycanohydrolase; EC 3.2.1.15].     

Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization

Lignocellulose is the major component of plant cell walls and it represents a great source of renewable organic matter. One of lignocellulose constituents is pectin. Pectin is composed of two basic structures: a ‘smooth’ region and a ‘hairy’ region. The ‘smooth’ region (homogalacturonan) is a linear polymer of galacturonic acid residues with α-(1→4) linkages, substituted by methyl and acetyl residues. The ‘hairy’ region is more complex, containing xylogalacturonan and rhamnogalacturonans I and II. Among the enzymes which degrade pectin (pectinases) is pectin lyase (E.C. 4.2.2.10). This enzyme acts on highly esterified homogalacturonan, catalysing the cleavage of α-(1→4) glycosidic bonds between methoxylated residues of galacturonic acid by means of β-elimination, with the formation of 4,5-unsaturated products. In this work, the gene and cDNA of a pectin lyase from Penicillium purpurogenum have been sequenced, and the cDNA has been expressed in Pichia pastoris. The gene is 1334 pb long, has three introns and codes for a protein of 376 amino acid residues. The recombinant enzyme was purified to homogeneity and characterized. Pectin lyase has a molecular mass of 45 kDa as determined by SDS-PAGE. It is active on highly esterified pectin, and decreases 40 % the viscosity of pectin with a degree of esterification ≥85 %. The enzyme showed no activity on polygalacturonic acid and pectin from citrus fruit 8 % esterified. The optimum pH and temperature for the recombinant enzyme are 6.0 and 50 °C, respectively, and it is stable up to 50 °C when exposed for 3 h. A purified pectin lyase may be useful in biotechnological applications such as the food industry where the liberation of toxic methanol in pectin degradation should be avoided.     

Evaluation of filamentous fungi and inducers for the production of endo-polygalacturonase by solid state fermentation

Aspergillus oryzae CCT 3940, Aspergillus awamori NRRL 3112 and a Trichoderma sp.) were compared for their capacity to produce endo-polygalacturonase (endo-PG) in solid state fermentation. Maximum pectinolytic activity was reached in 72 h of growth, the best two fungal strains being A. niger T0005007-2 and A. oryzae CCT 3940. Three types of commercial purified pectin and four of unprocessed pectin (tangerine, orange, Tahiti lime and sweet lime rind) were used to assess the effect of pectin on the production of endo-PG by A. niger T0005007-2. Maximum pectinolytic activity was achieved using 6 and 10% (w/w) of purified pectin as inducer. Depending on the origin of the commercial pectin used as inducer, maximum endo-PG levels varied from 223 to 876 units per gram of dry medium (one endo-PG unit (U) was defined as the quantity of enzyme which caused a reduction in viscosity of 50% in a 1% w/v solution of pectin in 30 min), indicating that care should be taken when choosing this component of the medium. When the crude pectins were used as inducers at the same concentration as purified pectin, maximum endo-PG activities were 250-300 units/g. However, by increasing the amount of Tahiti lime rind to 50% (w/w) maximum endo-PG was 919 U/g, thus opening up the possibility of a low cost medium for endo-PG production.     

Dried sugar beet pulp as a high energy feed for beef cattle

Digestibility trials with 8 lots of pelleted dried sugar beet pulp, carried out with wethers, demonstrated that dried sugar beet pulp is a highly digestible energy source for ruminants. From the digestion coefficients obtained, and a value number of 95, the starch equivalent of the dry matter of dried sugar beet pulp was calculated to be 73.3, while the net energy expressed in EF_r was 619 EF_r per kg dry matter. This is about 90% of the net energy content of barley containing 4% crude fibre in the dry matter.Beef production trials were carried out with 702 young bulls fed on complete dry rations based on dried sugar beet pulp. Two categories of animals were used: 322 baby-beef bulls (intensive system) slaughtered at 13 months of age at an average live-weight of 480 kg; and 380 young bulls coming from pasture (semi-intensive system) at about 250 kg live-weight, and fattened indoors up to at least 550 kg live-weight. With each category, three different rations have been studied. These contained respectively, 50, 60 or 70% pelleted dried sugar beet pulp; the remainder of the rations consisted of respectively 50, 40 or 30% concentrates. The diets were fed ad libitum; straw and water were always available. The three complete dry rations proved to be equally successful for intensive beef production. The carcass quality was good for all animals. The average daily gain obtained with the baby-beef bulls for the three rations respectively was 1 207 g, 1 274 g and 1 172 g; for the second category of bulls the mean growth rates were generally slightly higher: 1 281 g, 1 309 g and 1 357 g.The feed efficiency was higher with the younger animals: the baby-beef bulls (live-weight interval: 150–480 kg) consumed about 2.5 kg protein supplement and 3.5 kg dried sugar beet pulp per kg live-weight gain; while the intake per kg live-weight gain with the bulls of the second category (live-weight interval: 250–560 kg) amounted approximately to 2.75 kg protein supplement and 4 kg dried sugar beet pulp. Within each category of bulls, the feed cost per kg live-weight gain decreased with increasing amounts of dried sugar beet pulp in the rations.