Preparation and properties of reduced chitooligomers

In order to prevent the browning of the chitooligomer product prepared from enzymatic hydrolysis, the chitooligomers were reduced by potassium borohydride. The chemical structures and physical properties of the reduced chitooligomers were characterized by magnetic resonance spectra, X-ray diffraction, UV-spectrophotometry, thermogravimetric analysis, differential thermal analysis. The chemical stability of the chitooligomers increased after reduction. The reduced chitooligomers had no oral acute toxicity, and they had almost the same inhibition effect against the sarcoma 180 tumor cells in mice by intraperitoneal injection as the fresh chitooligomers.     

High polyphenol oxidase activity and low titratable acidity in browning bamboo tissue culture

Summary Tissue browning that frequently results in the early death of bamboo shoots in vitro correlated directly with polyphenol oxidase (PPO, EC 1.10.3.1) activity and inversely with titratable acidity. It was unrelated to the level of endogenous phenols. During the course of culture, timing of PPO activity paralleled that of explant browning. Browning was highest among shoots cultured in a medium of pH 8, which was consistent with the pH optinum of the bamboo enzyme. The pH optimum was first determined with the crude enzyme, then verified with two purified isozymes. Stability of the bamboo PPO was also highest at pH 10. PPO activities of the severely browning Dendrocalamus latiflorus, the moderately browning Phyllostachys nigra, and the relatively non-browning Bambusa oldhamii were inhibited strongly by ascorbic acid, cysteine, sodium diethyldithiocarbamate, and sodium sulfite. But characterization of bamboo PPO according to enzyme inhibitors was not possible because enzyme extracts of the three species gave varied responses to the traditional substances. Nutrient medium addenda of some PPO inhibitors, namely ascorbic acid, cysteine, kojic acid, and thiourea, mainly enhanced browning. However, ferulic acid at 3 mM and lower concentrations reduced the number of brown shoots per culture, although not the percentage of cultures that browned. Polyvinylpyrrolidone failed completely to suppress browning. The two purified isozymes showed different temperature optima for PPO activity: 60°C and 65°C. The purified isozymes displayed a substrate preference for dopamine, or a cathecol oxidase characteristics.     

Enzymatic rearrangement of chitine hydrolysates with β-N-acetylhexosaminidase from Aspergillus oryzae

The role of higher chitooligomers in medical applications is increasing due to their interesting biological activities. Transglycosylation activity of β-N-acetylhexosaminidase from Aspergillus oryzae was employed to produce higher chitooligosaccharides (chitohexaose–chitooctaose) from a mixture of lower chitooligomers prepared by acid hydrolysis of chitin. Enzymatic rearrangement of the chitooligomer mixture was optimized in respect of substrate concentration, presence of inorganic salts, enzyme activity, and reaction time to achieve the highest production of longer chitooligomers.     

Contributions of Amino-carbonyl Reaction,Caramelization and Some Other Reaction to Browning of Saké

In the previous report, it has been concluded that the browning of Saké involves three kinds of browning reactions which are the amino-carbonyl reaction, the caramelization and some other browning reaction by components other than glucose. In the present work, the extents of contributions of the three kinds of reactions to the browning of Saké were actually estimated, respectively, by analysing its dependence on the glucose concentration. It was concluded that the browning by components other than glucose was the major factor while the amino-carbonyl reaction and the caramelization were the minor ones in the browning of Saké. In addition, the change in pH value, as well as in temperature, was observed to influence exponentially on the browning rate of Saké. An empirical equation was proposed to elucidate the way in which the three kinds of reactions contributed to the overall browning in Saké.     

龙眼果实采后失水果皮褐变与活性氧及酚类代谢的关系

研究了(10±1)℃和50%相对湿度贮藏条件下"福眼"龙眼果实果皮褐变与活性氧和酚类代谢的关系.结果表明,采后失水导致龙眼果实果皮褐变,果皮活性氧清除酶SOD、CAT、APX、GR活性和内源抗氧化物质AsA、GSH含量下降,O-2产生速率和MDA含量增加,细胞膜透性迅速增大;PPO和POD活性增加,总酚和类黄酮含量明显下降.据此认为,果皮褐变可能是细胞的活性氧代谢失调,细胞膜结构破坏,使PPO、POD与酚类物质(含类黄酮)接触、酚类物质氧化的结果.     

Separation and scavenging superoxide radical activity of chitooligomers with degree of polymerization 6-16

The separation of chitooligomers (COS) with well-defined degree of polymerization (DP) is of interest to further study their bioactivity. However, there has been no report on separation of chitooligomers with DP>6 and the activity of these oligomers is unknown. This paper focuses on separating COS with DP>6 and five fractions were separated from the prepared fully deacetylated chitooligomers mixture by CM Sepharose Fast Flow column and analyzed by HPLC, which mainly contained glucosamine oligomers with DP6-7 (41.31%, 50.22%), DP7-8 (22.47%, 70.13%), DP9-10 (53.06%, 27.99%), DP10-12 (18.45%, 49.36%, 22.31%), and DP>12, respectively. The superoxide radical scavenging activity of each fraction was investigated. The oligomers with DP ranging from 10 to 12 exhibited higher scavenging activity than other fractions and in combination with the DP distribution of fractions, it was further concluded that the chitooligomers with DP11 was likely to be optimal for scavenging superoxide radical activity.     

蝴蝶兰离体培养及其相关生物技术研究进展

对蝴蝶兰(Phalaenopsis)的离体培养、褐化及控制、物理化学诱变、转基因研究进展进行了综述,包括蝴蝶兰离体培养中的外植体选择、体细胞胚胎及类原球茎诱导、培养基及培养条件、培养方式、褐化控制等关键因素,并概括了生物反应器、转基因、诱变育种等相关生物技术研究进展。成体植株的花梗、茎尖和种子以及试管苗的叶片、茎尖、根尖等均可作为外植体;影响外植体褐化的主要因素有培养基的种类、pH值、温度、外植体的生理状况等;蝴蝶兰诱变育种及转基因研究尚处于初步阶段。     

Preparation of Ultrafiltered Rice Bran Extract and Some Factors Affecting its Browning Inhibition in Potato Puree

Abstract

Rice bran extract (RBE) was fractionated by using ultrafiltration techniques. The obtained retentate (R) inhibited the browning of potato puree to a similar extent as RBE and more than it's permeate. The R that was incubated at pH 4.0, 5.0, and 6.0 inhibited the browning of potato puree more than R that was incubated at pH 7.0 or 8.0. Potato puree that was treated with heated R at 80°C for 10 min had a browning value similar to that of puree treated with unheated R and lower than those treated with heated R at 40 or 60°C for 10 min.     

Comparison of Keeping Quality between Pressure-processed Jam and Heat-processed Jam: Changes in Flavor Components,Hue, and Nutrients during Storage

Products of strawberry jam prepared by a high hydrostatic pressure-processing method and a conventional heat-processing method were examined with regard to the changes of quality such as volatile flavor components, anthocyanins, browning index, furfural, sucrose contents, and vitamin C contents during storage at 5°C and 25°C for 1–3 months. The quality of pressure-processed jam immediately after manufacturing was superior to that of heat-processed jam in preserving the fresh flavor and natural color of raw fruits. The superior quality was maintained under low temperature storage for two to three months. However, room temperature storage resulted in rapid deterioration including off-flavor, discoloration, browning, and decomposition of sucrose and vitamin C. So, the commercial value of pressure-processed jam decreased within a brief period. On the other hand, heat-processed jam did not change in quality under room temperature storage for three months. It is supposed that dissolved oxygen and enzyme systems retained are the causative factors of rapid deterioration in pressure-processed jam.     

Effects of Chemical Components on the Browning of Tomato Juice

The effects of eleven components in tomato juice on browning by heating were examined at the concentrations found in juice, About 70% of the browning of tomato juice was attributable to three constituents; fructose, galacturonic acid and glutamic acid. The interaction between glutamic acid and galacturonic acid or fructose was also found to accelerate the browning of tomato juice. The browning by heating of galacturonic acid with or without amino acids was larger than that of sugars or amino acids. The browning mechanism of galacturonic acid on heating at high temperature was discussed.     

Response surface approach to optimize temperature,pH and time on antioxidant properties of wild bush (Plectranthus rugosus) honey from high altitude region (Kashmir Valley) of India

In this study, the combined effect of temperature (60 to 80 °C) time (10 to 15 min.) and pH (3 to 6) was employed on the anti-oxidant potential (1,1-diphenyl-2-picrylhydrazyl-radical scavenging activity-DPPH-RSA, total phenolic content-TPC, and total flavonoid content-TFC) of wild bush Indian honey from high altitudes of Kashmir Valley by using response surface methodology (RSM). The statistical analysis showed that all the process variables had a substantial effect on the responses related to DPPH-RSA, TFC, and TPC, all of which increased as temperature and time increased. With an increase in pH, the antioxidant activity of wild bush honey was significantly decreased. The heat treatment of honey at high temperature (80 °C) was found to be more efficacious than at 70 and 60 °C, respectively. The findings showed that at higher temperature, browning pigments were formed which enhanced considerably the antioxidant activity of honey.     

Preparation of a series of chitooligomers and their effect on hepatocytes

A series of chitooligomers with molecular weight ranging from 1.7  3.8 × 10³ were prepared by degradation of a high molecular weight chitosan with hydrogen peroxide and selective precipitation in ethanol solutions. The prepared chitooligomers were characterized by gel permeation chromatography, elemental analysis, Fourier transform infrared spectra, ¹³C nuclear magnetic resonance spectroscopy and X-ray diffraction analysis. Cell culture experiments suggested that the effect of the chitooligomers on the proliferation of L02 hepatocytes was dependent on culture time, namely, at the initial stage of culture there was an inhibitory effect on proliferation of the cells; however, the cultures recovered in cell proliferation and exhibited promotion effect in following days. In the case of chitosan monomer (GlcN), high concentration of GlcN (1 mg/ml) produced a significant suppression in proliferation of L02 cells relative to control, with decreases of 35.2%, 60% and 72.9% on days 1, 3 and 5, respectively. In addition, there was no significant effect of the chitooligomers on the functions of albumin secretion and urea synthesis of the hepatocytes.     

The oxygen uptake of Sarotherodon niloticus L. and the oxygen binding properties of its blood and hemolysate (Pisces: Cichlidae)

The oxygen consumption of Sarotherodon niloticus L. was found to decline below a critical oxygen concentration of about 2 mg O2/l. An important influence of CO2 on the oxygen affinity of whole blood was observed at all temperatures between 20 and 35 degrees C for gas mixtures containing 5.6% CO2. Purified hemolysate showed extremely high oxygen affinities (p50 = 1.08 mmHg at pH 8.2 and 20 degrees C). Low cooperativity was observed at all temperatures from 20 to 35 degrees C, and pH values between 6.5 and 8.2. The Bohr effect proved to be important at pH values lower than pH 7.5 (phi = delta log P50/delta pH = -0.58 between pH 6.5 and 7.0 at 35 degrees C). The oxygen affinities show high thermal sensitivity without a marked pH influence (delta H value for overall oxygenation at pH was -71.7 kJ/mol). The obtained results are interpreted as adaptations to diurnal variations in ambient temperature and oxygen availability.     

环境因子对卵形鲳鲹幼鱼耗氧率和排氨率的影响

运用封闭流水式实验方法研究温度、盐度、pH和流速对卵形鲳鲹(Trachinotus ovatus)幼鱼耗氧率和排氨率的影响.实验结果表明,随着温度的升高,耗氧率和排氨率均是先增大后减小,当温度为27℃时,耗氧率和排氨率达最大值,温度对卵形鲳鲹幼鱼耗氧率和排氨率的影响显著(P＜0.0l);耗氧率和排氨率随着盐度的升高均出...     

Hemoglobins of the tadpole of the bullfrog, Rana catesbeiana. Temperature dependence of oxygen binding and pH dependence of subunit dissociation.

The temperature dependence of the oxygen equilibrium of tadpole hemoglobin has been determined between 0 degrees and 32 degrees for the unfractionated but phosphate-free lysate and between 12 degrees and 32 degrees for each of the four isolated components between pH 6 and 10 in 0.05 M cacodylate, Tris, or glycine buffers containing 0.1 M NaCl and 1 mM EDTA. Under these conditions the Bohr effect (defined as deltalog p50/deltapH) of the unfractionated lysate is positive at low temperatures between pH 6 and 8.5 and is negative above pH 8.5 to 8.8 at any temperature. As the temperature rises the Bohr effect below pH 8.5 changes greatly. In the interval pH 7.0 to 7.5, the magnitude of the Bohr effect decreases from + 0.28 at 0 degrees to zero at about 24 degrees and becomes negative, as in mammalian hemoglobins, above this temperature. Measurements with the isolated components show that the temperature dependence of oxygen binding for Components I and II and for Components III and IV is very similar. For both sets of components the apparent overall enthalpy of oxygenation at pH 7.5 is about -16.4 kcal/mol and -12.6 kcal/mol at pH 9.5. The measured enthalpies include contributions from the active Bohr groups, the buffer ions themselves, the hemoglobin groups contributing buffering, and any pH-dependent, oxygenation-dependent binding of ions such as chloride by the hemoglobin. The apportioning of the total enthalpy among these various processes remains to be determined. Between pH 8 and 10.5 tadpole oxyhemoglobin undergoes a pH-dependent dissociation from tetramer to dimer. The pH dependence of the apparent tetramer-dimer dissociation constant indicates that at pH 9.5 the dissociation of each tetramer is accompanied by the release of approximately 2 protons. In this pH range the oxygen equilibrium measurements indicate that about 0.5 proton is released for each oxygen molecule bound. The results are consistent with the conclusion that one acid group per alphabeta dimer changes its pK from about 10 to 8 or below upon dissociation of the tetramer.     

Characterization and kinetics of 45 kDa chitosanase from Bacillus sp. P16

An extracellular 45 kDa endochitosanase was purified and characterized from the culture supernatant of Bacillus sp. P16. The purified enzyme showed an optimum pH of 5.5 and optimum temperature of 60 degrees C, and was stable between pH 4.5-10.0 and under 50 degrees C. The Km and Vmax were measured with a chitosan of a D.A. of 20.2% as 0.52 mg/ml and 7.71 x 10(-6) mol/sec/mg protein, respectively. The enzyme did not degrade chitin, cellulose, or starch. The chitosanase digested partially N-acetylated chitosans, with maximum activity for 15-30% and lesser activity for 0-15% acetylated chitosan. The chitosanase rapidly reduced the viscosity of chitosan solutions at a very early stage of reaction, suggesting the endotype of cleavage in polymeric chitosan chains. The chitosanase hydrolyzed (GlcN)7 in an endo-splitting manner producing a mixture of (GlcN)(2-5). Time course studies showed a decrease in the rate of substrate degradation from (GlcN)7 to (GlcN)6 to (GlcN)5, as indicated by the apparent first order rate constants, k1 values, of 4.98 x 10(-4), 2.3 x 10(-4), and 9.3 x 10(-6) sec(-1), respectively. The enzyme hardly catalyzed degradation of chitooligomers smaller than the pentamer.     

抗氧化剂对苜蓿原生质体早期培养的影响

研究发现,分离原生质体的酶解脱壁处理可以诱导苜蓿细胞产生活性氧。培养基中添加抗氧化剂,有助于提高培养原生质体的分裂频率,缓解褐化现象的出现。经紫外照射处理的培养基不利于苜蓿原生质体的生长和分裂,添加抗氧化剂后,紫外辐射所引起的不良效应则被抵消。因而,通过抗氧化剂对活性氧的清除,有助于早期原生质体的培养。     

The catalytic potency of β-glucosidase from Pyrococcus furiosus in the direct glucosylation reaction

Enzymes from extremophiles operate at conditions that are different from their ‘normal’ counterparts, and are therefore a useful extension of the enzyme toolbox. In this paper, the direct glucosylation reaction mediated by a hyperthermophilic β-glucosidase from Pyrocuccus furiosus was investigated. Hexanol was successfully coupled to glucose with this enzyme. A preliminary study was conducted to improve the product yield. A maximum product concentration of 12.9 g.l⁻¹ was attainable by increasing the glucose concentration to the maximum solubility of 2000 g.(kg buffer solution)⁻¹ at the reaction temperature. The highest glucose based yield of 2.64% was achieved with a glucose concentration of 900 g.(kg buffer solution)⁻¹ at a reaction temperature of 65°C and a pH of 6.0. Performing the reaction at higher pH and temperature led to lower product concentrations. This was caused by deactivation of the enzyme accompanied by browning of the reaction mixture. A pH of 4.4 did have a negative effect on both the storage and the operational stability of the enzyme.     

Effect of different soil types,pH and phosphorus levels on the wheat seedling and the incidence ofPythium graminicolum

Summary In the present study, the effect of different types of soil, pH and phosphorus levels on root, shoot development and browning extent on shoot of wheat seedlings growth in soil inoculated withPythium graminicolum; was studied. Out of the four soil types the development of root and shoot was better in Domatta soil where as the disease development was more in black clay soil. The pH values below 5.5 and above 8.0 are favourable for the root and shoot development whereas these values are unfavourable for the development of the Pythium and consequently for infection by it. The percentage of browning of the collar region was more in soils near neutral (pH 6.5–7.1) and it was less in acidic (pH 5.0) or alkaline (pH 8.6) soils. The development of wheat roots and shoot in soil with 30 lb/acre phosphorus was good as compared to that in soil with P₀, P_15,45 and P₆₀ lb/acre in inoculated soil. The percentage of extent of browning on collar region was less at 15 lb and 30 lb/acre P, due to less development of the disease.     

Oxygen is not required for the browning and crosslinking of protein by pentoses: relevance to Maillard reactions in vivo

The chemical modification and crosslinking of proteins by the Maillard or browning reaction contributes to the aging of tissue proteins, and acceleration of this reaction during hyperglycemia is implicated in the pathogenesis of diabetic complications. Metal-catalyzed autoxidation reactions catalyze the browning of proteins by glucose, a process known as autoxidative glycosylation, but the effects of oxidative conditions on browning of proteins by smaller sugars has not been reported. In this work we studied the browning and crosslinking of the model protein, RNase A, by pentoses. Although antioxidative conditions inhibited the formation of glyoxal and the advanced glycation end-product, N epsilon-(carboxymethyl)lysine from arabinose, browning and crosslinking, and formation of the fluorescent crosslink pentosidine proceeded at comparable rates under oxidative and antioxidative conditions. These studies and other work on smaller dicarbonyl compounds indicate that Maillard reactions of simpler carbohydrates proceed efficiently in the absence of oxygen and suggest that antioxidant therapy for treatment of diabetic complications may have limited clinical efficacy.