红酵母属中的一个新种

自云南省哀牢山常绿阔叶林根际土壤中分离出一株酵母菌Ⅱ-33。该菌株细胞卵圆,出芽时胞壁结构为内分芽型,多端芽殖,不产生子囊孢子和掷孢子;在麦芽汁琼脂上产生红色胡萝卜素,颜色反应阳性,菌落平滑,有光泽,质地软而呈粘液状,不发酵任何糖,能同化麦芽糖、蔗糖、乳糖、棉子糖、松三糖和硝酸钾,能分解尿素;细胞水解物中含甘露糖、葡萄糖和半乳糖。按其形态、培养特征和生理生化特性,酵母菌Ⅱ-33应归入红酵母属(Rhodotorula)中,经与近似种比较,认为是红酵母属中的一个新种,命名为云南红酵母(Rhodotorula yunnanensis sp.n.)。     

鸡胚凝集因子在发育过程中的活性变化

人们认为,细胞表面含碳水化合物的蛋白质可能在细胞相互作用中起着重要作用。从脊椎动物中分离出凝集因子以后(Teichberg et al., 1975;Nowak et al., 1977),许多研究表明这类凝集因子是与膜相结合的。这些发现支持了人们原先的设想。Beyer等人(1979;1980)从成体鸡肠中分离出一种结合乳糖的凝集因子,这种凝集因子似乎存在于粘液分泌小粒之中。1980年Pitts和Yang又从鸡胚肾脏中分离出一种结合乳糖的凝集因子,它可能是一种外周蛋白质。尽管他们的实验证明了成体鸡肾和肠这两种器官,同视网膜、肝脏、肌肉、心脏、脑和脊髓一样,存在着结合乳糖的凝集因子。但凝集因子在这两     

SIGNIFICANCE OF THE CHEMICAL COMPOSITION OF THE SECRETING AND DRY MAMMARY GLAND TO MILK SECRETION

The results herein presented furnish exact critical evidence for one more stage in milk secretion. Cows producing up to 30 pounds of milk at one milking are shown to have the lactose equivalent of all this milk in the udder when milking commences. The average excess of lactose found in the udder after subtracting the amount necessary for the contained milk is equal to 2.1 pounds. This represents the milk retained in the udder when the cow is believed to be dry. These conclusions are further supported by the fact that no sugar is found in the udder in the quiescent state. The study of the total composition of the udder as fat, ash, nitrogen, and lactose, and of the contained milk shows that there is a large excess of fat, ash, and nitrogen in proportion to that necessary for milk formation. The excess of udder lactose over the milk lactose is much less. The lactose would therefore appear to be formed from some element in the blood, probably dextrose, only as needed for the formation of milk. The composition of the dry udder is quite different in certain respects from that of the actively secreting gland. It builds up a fat reserve of a quite different Reichert-Meissl number from that of butter-fat. It has no sugar, its ash content is reduced, and the nitrogen content is like that of the secreting gland.     

柑桔园土壤脲酶活性年周期变化及脲酶抑制剂氢醌的效应

本研究初步探明南亚热带赤红壤柑桔园土壤脲酶活性的年周期变化:冬季最低,春季升至高峰,夏季开始下降,直至秋季仍处于较低水平。室内模拟试验表明:氢醌对柑桔园土壤脲酶活性有明显抑制作用,其抑制率随浓度提高而增加,且随着培养时间的延长而逐渐下降;添加氢醌处理的尿素水解持续时间,比不加氢醌的延长约1倍。     

Energy dependence of different steps in the autophagic-lysosomal pathway

The energy dependence of the autophagic-lysosomal pathway was investigated in isolated rat hepatocytes, using electroinjected [14C]lactose as an autophagy probe and atractyloside to alter intracellular ATP levels. Since autophagocytosed lactose is hydrolyzed in lysosomes, several steps in the pathway could be analyzed. The following observations were made. 1) The overall autophagic degradation of electroinjected [14C]lactose was strongly energy-dependent. More than 85% inhibition was obtained when the ATP content decreased from the control value of 10 mumol/g dry weight to 4 mumol/g dry weight. 2) The initial step, i.e. the autophagic sequestration of [14C]lactose, measured in the presence of vinblastine to prevent transfer of lactose to lysosomes, was as sensitive to small changes in ATP as was the overall lactose degradation. 3) The steady state level of sequestered [14C]lactose remained constant as ATP decreased from 10 to 4 mumol/g dry weight, indicating that the sequestration step and some postsequestrational process were inhibited to a similar extent by ATP depletion. 4) The final step in the pathway, intralysosomal hydrolysis, was measured by allowing [14C]lactose to preaccumulate intralysosomally in the presence of the reversible lysosome inhibitor propylamine. Following propylamine removal and inhibition of further sequestration by 3-methyladenine, ATP-dependent hydrolysis of the intralysosomal [14C]lactose could be demonstrated. However, this hydrolysis step was not as sensitive to small changes in ATP as was the sequestration step or the overall autophagic lactose degradation. Control of the autophagic-lysosomal pathway in response to energy deprivation would therefore not seem to occur at the lysosomal level, but may be exerted both at the sequestration step and at a postsequestrational, prelysosomal step.     

Adaptation of a manometric biosensor to measure glucose and lactose

A manometric sensor previously developed to measure urea was modified to measure glucose and lactose through enzymatic oxidation. Change in pressure in an enclosed cavity was correlated to the depletion of oxygen resulting from the enzymatic oxidation of glucose or lactose. The response of the sensor was linear and could be made adjustable over a large range by adjusting the amount of sample loaded into the fixed volume reactor. Because of the slow mutarotation of glucose, the oxidation of glucose was not allowed to proceed to completion. Therefore, the precision of the sensor (approximately 0.2 mM in a range from 0 to 5 mM) was limited by variations in the oxidation rate of glucose by glucose oxidase. Because the assay for lactose measured glucose subsequent to the hydrolysis of lactose by beta-galactosidase, the same degree of precision was observed in lactose. Milk lactose, typically at concentrations of about 150 mM, was estimated using the lactose assay after first diluting the samples. For many fluids such as milk, the use of manometric sensors for oxidizable substrates may be preferable to optical and electrochemical methods because they are robust and suffer a low degree of optical and chemical interferences. Glucose and lactose are representative of many important oxidizable substrates, which may be determined in this manner, many of which do not suffer from limitations caused by mutarotation. In theory, detection limits less than 1 microM may be achieved using these methods.     

Transferase reactions of the β-galactosidase from Streptococcus thermophilus

Summary The -galactosidase from Streptococcus thermophilus formed transferase products (including up to six disaccharides and two trisaccharides) during the hydrolysis of lactose to glucose and galactose. The extent of transferase products formed was dependent on the initial lactose concentration, reaching up to 40% of the total carbohydrate at 70% w/v lactose. At high lactose concentrations (40% w/v) trisaccharide transferase products were formed initially, followed by the appearance of disaccharide transferase products. In contrast, at low lactose concentrations (7.5 w/v), only traces of the trisaccharides were detected with disaccharides being the predominant transferase products. The disaccharide products accumulated to relatively high concentrations late in the overall hydrolysis of lactose, at both high and low initial lactose concentrations, while the trisaccharides peaked much earlier and were themselves subsequently hydrolysed prior to the complete disappearance of lactose. It was possible to study the hydrolysis of galactosyl lactose by the S. thermophilus -galactosidase using a semi-pure galactosyl lactose preparation containing 5% lactose. The hydrolysis of this trisaccharide occurred via at least four disaccharide intermidiates, which appeared chromatographically identical to the disaccharide transferase products formed during lactose hydrolysis. This suggests that the enzymic formation and subsequent hydrolysis of galactosyl lactose occurs via coincident reaction pathways. The initial rate of galactose over glucose formation during galactosyl lactose hydrolysis changed from a ratio of 3:1 at low (2–3% w/v) substrate concentrations to 1.5:1 at high (>20% w/v) concentrations. This indicates a shift in the preferred initial cleavage site from the galactose-galactose bond to the galactose-glucose bond.     

Enzymes involved in carbohydrate metabolism and their role on exopolysaccharide production in Streptococcus thermophilus

The role of the enzymes uridine-5'-diphospho-(UDP) glucose pyrophosphorylase and UDP galactose 4-epimerase in exopolysaccharide production of Gal⁻ ropy and non-ropy strains of Streptococcus thermophilus in a batch culture was investigated. Growth of the ropy and non-ropy strains was accompanied by total release of the galactose moiety from lactose hydrolysis in modified Bellinker broth with lactose as the only carbon source. This was associated with a greater exopolysaccharide production by the ropy strain. The polymer produced by both strains in cultures with lactose or glucose as carbon sources contained glucose, galactose and rhamnose, indicating that glucose was used as a carbon source for bacterial growth and for exopolysaccharide formation. UDP-glucose pyrophosphorylase activity was associated with polysaccharide production during the first 12 h in a 20 h culture in the ropy strain, but not in the non-ropy strain. UDP-galactose 4-epimerase was not associated with exopolysaccharide synthesis in any strain. The evidence presented suggests that the glucose moiety from lactose hydrolysis is the source of sugar for heteropolysaccharide synthesis, due to a high UDP-glucose pyrophosphorylase activity.     

Formation of galacto-oligosaccharides during lactose hydrolysis by a novel beta-galactosidase from the moderately thermophilic fungus Talaromyces thermophilus

Discontinuous and continuous processes of lactose hydrolysis and concomitant galacto-oligosaccharide (GalOS) formation were studied. To this end a wide experimental range of the main variables was evaluated, including the initial lactose concentration, the degree of lactose conversion, the pH value and the temperature for discontinuous transformations, while the initial lactose concentration and the feed rate were varied for the continuous process. For both processes a high-initial lactose concentration proved to be advantageous for the formation of GalOS. The maximum amount of GalOS (100 g/L, corresponding to a yield of approximately 50% based on the initially employed lactose) was obtained after 8 h of incubation when using 200 g/L lactose as substrate and 90% lactose hydrolysis was observed. GalOS productivity in the continuous process (g/L.h) was enhanced by an increase of the flow rate. The maximum GalOS productivity of 70 g/L.h was obtained at a flow rate of 24 mL/h when using a reactor with a total working volume of 21 mL. As was evident from these experiments, this beta-galactosidase from a moderately thermophilic fungus showed a strong transgalactosylation activity and can be used for the formation of GalOS, sugars that are of considerable interest for functional food applications because of their presumed healthpromoting effects.     

STUDIES ON THE LACTASE OF ESCHERICHIA COLI

A "lactase solution" was prepared from Escherichia coli. The mechanism of its action has been studied and changes in the rate of hydrolysis under various conditions investigated. The hydrolysis of lactose by the enzyme approximates the course of reaction of the integrated Michaelis-Menten equation. One molecule of enzyme combines with one molecule of substrate. E. coli lactase is readily inactivated at pH 5.0, and its optimal activity at 36°C. is reached between pH 7.0 and pH 7.5. The optimal temperature for its action was found to be 46°C. when determinations were carried out after an incubation period of 30 minutes. Its inactivation by heat follows the course of a first order reaction, and the critical thermal increment between the temperatures of 45°C. and 53°C. was calculated to be 56,400 calories per mol. The enzyme is activated by potassium cyanide, sodium sulfide, and cysteine, and irreversibly inactivated by mercuric chloride, silver nitrate, and iodine. After inactivation with copper sulfate partial reactivation is possible, while the slight inhibition brought about by hydrogen peroxide is completely reversible. The possible structure of the active groups of E. coli lactase as compared with other enzymes has been discussed.     

灭幼脲Ⅲ号在好气水环境中的降解代谢的初步研究

本文研究了新农药灭幼脲Ⅲ号在好气水环境中的降解与代谢。在避光条件下,观察了灭菌组与实验组中灭幼脲Ⅲ号及其主要代谢产物的消长过程,比较了它的化学水解与微生物降解的差异。在室内模拟好气系统中,研究了母体化合物在水体中的残留动态和生物降解半衰期,及其初期主要代谢途径的转化产物,同时分别用高效液相色谱法,紫外吸收光谱扫描,以及特征有机质谱图,对灭幼脲Ⅲ号的两种主要代谢产物进行了定性定量测定。结果表明:灭幼脲Ⅲ号在室内好气环境中较易水解,而且水中微生物的存在能加速它的降解,母体化合物在水体中初期代谢主要途径为分子中的苯甲酰碳与脲氮键首先开裂,生成邻氯苯甲酸(CBA)和对氯苯基脲素(CPU)。     

Rate of hydrolysis of urea as influenced by thiourea and pellet size

Summary Two incubation experiments and a number of field experiments were conducted to determine the effect of soil moisture tension, pellet size and addition of thiourea to urea on the rate of urea hydrolysis. In the incubation experiments at 20°C, the rate of hydrolysis of urea increased from 15 bar to 1/3 bar soil moisture tension, with the largest change (doubling) occurring from 15 bar to 7 bar moisture tension. Increasing pellet size reduced the rate of urea hydrolysis by about 12% with urea pellets weighing 0.21 g as compared to 0.01 g urea pellets after 114h. When thiourea (a metabolic inhibitor) was pelleted with urea in a ratio of two parts urea and one part thiourea, the rate of hydrolysis was halved.In a field experiment, the addition of thiourea to urea and increasing pellet size suppressed the rate of urea hydrolysis considerably for 8 days. The amount of urea hydrolyzed with urea+thiourea (21) pellets weighing 2.51 g was one-fourth of the amount of urea hydrolyzed with 0.01 g pellets of urea alone. In the other six field experiments which were set out in October, only 22% to 39% of urea +thiourea (21) was hydrolyzed at two weeks after application, while almost all of the urea was hydrolyzed when it was mixed into the soil without an inhibitor.Unter our field conditions, we would estimate that the hydrolysis of urea can be inhibited for at least one week. The inhibition of urea hydrolysis appears to be great enough that the problems encountered from the rapid hydrolysis of urea, wherever these occur, may be reduced by combined use of thiourea and either increased pellet size or band placement.     

THE R?LE OF THE ACTIVITY COEFFICIENT OF THE HYDROGEN ION IN THE HYDROLYSIS OF GELATIN

1. The hydrolysis of gelatin at a constant hydrogen ion concentration follows the course of a monomolecular reaction for about one-third of the reaction. 2. If the hydrogen ion concentration is not kept constant the amount of hydrolysis in certain ranges of acidity is proportional to the square root of the time (Schütz''s rule). 3. The velocity of hydrolysis in strongly acid solution (pH less than 2.0) is directly proportional to the hydrogen ion concentration as determined by the hydrogen electrode i.e., the "activity;" it is not proportional to the hydrogen ion concentration as determined by the conductivity ratio. 4. The addition of neutral salts increases the velocity of hydrolysis and the hydrogen ion concentration (as determined by the hydrogen electrode) to approximately the same extent. 5. The velocity in strongly alkaline solutions (pH greater than 10) is directly proportional to the hydroxyl ion concentration. 6. Between pH 2.0 and pH 10.0 the rate of hydrolysis is approximately constant and very much greater than would be calculated from the hydrogen and hydroxyl ion concentration. This may be roughly accounted for by the assumption that the uncombined gelatin hydrolyzes much more rapidly than the gelatin salt.     

Hydrolysis of milk lactose by immobilized beta-galactosidase-hen egg white powder

Immobilized beta-galactosidase was obtained by crosslinking the enzyme with hen egg white using 2% glutaraldehyde. The gel obtained could be lyophilized to give a dry enzyme powder. The pH optimum of both the soluble and immobilized enzyme was found to be 6.8. The immobilized enzyme showed a higher K(m) for the substrates. The extent of enzyme inhibition by galactose was reduced upon immobilization. The stability towards inactivation by heat, urea, gamma irradiation, and protease treatment were enhanced. The bound enzyme as tested in a batch reactor could be used repeatedly for the hydrolysis of milk lactose. The possible application of this system for small-scale domestic use has been suggested.     

THE TEMPERATURE COEFFICIENT OF THE UREA DENATURATION OF EGG ALBUMIN

Evidence is brought forward to show that at concentrations of urea high enough to split the egg albumin molecule the solubility changes produced by urea are profoundly modified. The degree of precipitation after dialysis is the net result of two changes produced by the urea: the first, normally spoken of as denaturation, which makes the protein insoluble in dilute solution and the second, a splitting of the molecule which makes it soluble. These two reactions may proceed independently and simultaneously or the second reaction may follow the first, taking place in the denatured molecule only. In view of the decrease in the opalescence with time, the latter process is more probable. Both of these reactions have positive temperature coefficients, but as the concentration of urea increases the second reaction is more affected by increase in temperature than the first, and consequently the resulting opalescence decreases rather than increases with temperature. This accounts for and explains reports of negative temperature coefficients of denaturation, when denaturation is measured by the amount of insoluble material found on dilution. The occurrence of these two reactions, one leading to an increase and the other to a decrease in the amount of insoluble protein, should be taken into account when denaturation changes in egg albumin with urea are studied.     

β-Galactosidase from a cold-adapted bacterium: purification, characterization and application for lactose hydrolysis

The enzyme beta-galactosidase was purified from a cold-adapted organism isolated from Antarctica. The organism was identified as a psychotrophic Pseudoalteromonas sp. The enzyme was purified with high yields by a rapid purification scheme involving extraction in an aqueous two-phase system followed by hydrophobic interaction chromatography and ultrafiltration. The beta-galactosidase was optimally active at pH 9 and at 26 degrees C when assayed with o-nitrophenyl-beta-D-galactopyranoside as substrate for 2 min. The enzyme activity was highly sensitive to temperature above 30 degrees C and was undetectable at 40 degrees C. The cations Na+, K+, Mg2+ and Mn2+ activated the enzyme while Ca2+, Hg2+, Cu2+ and Zn2+ inhibited activity. The shelf life of the pure enzyme at 4 degrees C was significantly enhanced in the presence of 0.1% (w/v) polyethyleneimine. The pure beta-galactosidase was also evaluated for lactose hydrolysis. More than 50% lactose hydrolysis was achieved in 8 h in buffer at an enzyme concentration of 1 U/ml, and was increased to 70% in the presence of 0.1% (w/v) polyethyleneimine. The extent of lactose hydrolysis was 40-50% in milk. The enzyme could be immobilized to Sepharose via different chemistries with 60-70% retention of activity. The immobilized enzyme was more stable and its ability to hydrolyze lactose was similar to that of the soluble enzyme.     

Formation of oligosaccharides during enzymatic lactose: Part I: State of art

Enzymatic lactose hydrolysis by beta-galactosidase (lactase) was investigated with respect to the formation of oligosaccharides. An analysis of the formation of oligosaccharides and their control is important in the development of technical applications for enzymatic lactose hydrolysis. The available literature data on transfer reactions of lactase were reviewed, compared, and presented in a concise tabular form. Mechanisms and possible ways of modelling enzymatic lactose hydrolysis, including formation of oligosaccharides, are presented.     

Hydrolysis of lactose solutions and wheys by whole cells of Kluyveromyces bulgaricus

Summary Some factors affecting the hydrolysis of lactose solution and whey by whole cells of Kluyveromyces bulgaricus were studied. The K_m values were 59 mM and 32 mM for whole cells and cell-free extracts respectively. Optimum hydrolysis activity was observed at 48°C. At this temperature, 80% hydrolysis was obtained in lactose solution and whey after 3.5 and 9 min respectively by yeast cells (15 mg/ml) with an activity of 1.13 U/mg. Protein concentration in whey did not have an inhibitory effect. In whey permeate, cells were reused eight times with a hydrolysis degree of more than 80% but in lactose phosphate buffer, the hydrolysing capacity was lost quickly.     

Cold active β-galactosidase from Thalassospira sp. 3SC-21 to use in milk lactose hydrolysis: a novel source from deep waters of Bay-of-Bengal

The cold active β-galactosidase from psychrophilic bacteria accelerate the possibility of outperforming the current commercial β-galactosidase production from mesophilic sources. The present study is carried out to screen and isolate a cold active β-galactosidase producing bacterium from profound marine waters of Bay-of-Bengal and to optimize the factors for lactose hydrolysis in milk. Isolated bacterium 3SC-21 was characterized as marine psychrotolerant, halophile, gram negative, rod shaped strain producing an intracellular cold active β-galactosidase enzyme. Further, based upon the 16S rRNA gene sequence, bacterium 3SC-21 was identified as Thalassospira sp. The isolated strain Thalassospira sp. 3SC-21 had shown the enzyme activity between 4 and 20?°C at pH of 6.5 and the enzyme was completely inactivated at 45?°C. The statistical method, central composite rotatable design of response surface methodology was employed to optimize the hydrolysis of lactose and to reveal the interactions between various factors behind this hydrolysis. It was found that maximum of 80.18?% of lactose in 8?ml of raw milk was hydrolysed at pH of 6.5 at 20?°C in comparison to 40?% of lactose hydrolysis at 40?°C, suggesting that the cold active β-galactosidase from Thalassospira sp. 3SC-21would be best suited for manufacturing the lactose free dairy products at low temperature.     

Modes of lactose uptake in the yeast species Kluyveromyces marxianus

Twelve lactose-assimilating strains of the yeast species Kluyveromyces marxianus and its varieties marxianus, lactis and bulgaricus were studied with respect to transport mechanisms for lactose, glucose and galactose, fermentation of these sugars and the occurrence of extracellular lactose hydrolysis. The strains fell into three groups. Group I (two strains): Fermentation of lactose, glucose and galactose, extracellular lactose hydrolysis, apparent facilitated diffusion of glucose and galactose; Group II (two strains): Lactose not fermented, glucose and galactose fermented and transported by an apparent proton symport, extracellular hydrolysis of lactose present (one strain) or questionable; Group III (eight strains): Lactose, glucose and galactose fermented, lactose transported by an apparent proton symport mechanism, extracellular hydrolysis of lactose and transport modes for glucose and galactose variable.