Water activity and substrate concentration effects on lipase activity

Catalytic activity of lipases (from Rhizopus arrhizus, Canadida rugosa, and Pseudomonas sp. was studied in organic media, mainly diisopropyl ether. The effect of water activity (a(w)) on V(max) showed that the enzyme activity in general increased with increasing amounts of water for the three enzymes. This was shown both for esterification and hydrolysis reactions catalyzed by R. arrhizus lipase. In the esterification reaction the K(m) for the acid substrate showed a slight increase with increasing water activities. On the other hand, the K(m) for the alcohol substrate increased 10-20-fold with increasing water activity. The relative changes in K(m) were shown to be independent of the enzyme studied and solvent used. The effect was attributed to the increasing competition of water as a nucleophile for the acyl-enzyme at higher water activities. In a hydrolysis reaction the K(m) for the ester was also shown to increase as the water activity increased. The effect of water in this case was due to the fact that increased concentration of one substrate (water), and thereby increased saturation of the enzyme, will increase the apparent K(m) of the substrate (ester) to be determined. This explained why the hydrolysis rate decreased with increasing water activity at a fixed, low ester concentration. The apparent V(max) for R. arrhizus lipase was similar in four of six different solvents that were tested; exceptions were toulene and trichloroethylene, which showed lower values. The apparent K(m) for the alcohol in the solvents correlated with the hydrophobicity of the solvent, hydrophobic solvents giving lower apparent K(m). (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 798-806, 1997.     

Effect of water activity on production and activity of Rhizopus oligosporus polysaccharidases

Summary During tempeh fermentation, Rhizopus oligosporus produced polysaccharidases to degrade soya bean cell walls; the maximum activity for all polysaccharidases tested occurred 20–30 h after inoculation. R. oligosporus was also grown in a soya bean extract model medium to which glycerol was added to control water activity (a _w). The overall activities of the major enzymes produced by the fungus, polygalacturonase, endocellulase and xylanase, appeared to be strongly influenced by a _w. The production of enzymes as well as their specific activities were affected by a _w. The optimum a _w for polygalacturonase and xylanase activity coincided with that for mycelial growth, namely 0.99–1.00. In contrast, the optimum a _w for (endo)cellulase was 0.98, at which mycelial growth was significantly reduced. Correspondence to: M. Sarrette     

Influence of memory demand and contra lateral activity on muscle activity.

Computer mouse work often includes memory demands and contra lateral activity. This study simulated video display unit (VDU) mouse-work and the focus was on forearm muscle activity as a result of standardised postural loads, memory demands and contra lateral activity. Surface and intramuscular electromyography (EMG) were recorded from the right forearm muscles during finger elevation and rest with and without memory demands and with and without contra lateral activity i.e. activity of the left hand. In most situations, memory demand increased activity in the m. extensor carpi radialis brevis and m. flexor digitalis superficialis. Also contra lateral activity increased activity in situations with and without memory demands. While surface EMG level of the m. extensor digitorum communis did not increase during memory demands, intramuscular EMG level increased when memory demands and contra lateral activity was combined. Influence of memory demands and contra lateral activity were most pronounced, in situations where activity levels were small.We presume that it is not only prolonged time of active computer mouse use that is a risk for development of musculoskeletal disorders, but also the time when people interact with the computer mentally or with the 'non-mouse hand', while resting their 'mouse-hand' on the mouse.     

Studies on cell-clearing activity in alpha-lactalbumin. Effects of calcium removal on activity and conformation.

Effects of calcium removal on the cell-clearing activity of alpha-lactalbumin (alpha-LA) and concomitant changes in conformational structure have been investigated as part of a continuing study of the activity found earlier [McKenzie, H.A. & White, F.H., Jr. (1987) Biochem. Int. 14, 347]. This activity is similar to that of lysozyme, whereby lysis of the bacterial cell wall is catalyzed. However, the specific activity of alpha-LA is on the order of 10(-6) that of lysozyme. Under conditions where activities of apo and native alpha-LA were approximately linear functions of the protein concentration, the maximal ratio of apo to native activity was 5.7:1, determined by comparison of second order velocity constants. The CD spectrum of apo alpha-LA is intermediate between that of the A state and the native protein. By NMR, the conformation of apo alpha-LA is similar to, but distinctly different from, that of the native protein. The apo form did not revert completely to the native state when Ca(II) was resupplied, consistent with a role for this cation in folding. It is suggested that the activity increase may result from a diminished constriction of the "cleft" region in alpha-LA.     

蛇床子素粉剂对玉米象成虫的杀虫活性及酶活性的影响

研究1.0%蛇床子素(osthole)粉剂对玉米象Sitophilus zeamais Motschusky成虫的杀虫活性及其对玉米象成虫羧酸酯酶、乙酰胆碱酯酶、谷胱甘肽S-转移酶及蛋白质活性的影响。结果表明,1.0%蛇床子素粉剂对玉米象成虫有很好的杀虫活性,处理玉米象成虫3d后,0.8mg/kg以上剂量的蛇床子素粉剂对玉米象成虫的校正死亡率达100%;用0.19mg/kg蛇床子素粉剂处理玉米象成虫2d后,试虫体内乙酰胆碱酯酶、羧酸酯酶和蛋白质活性均表现出受抑制,而谷胱甘肽S-转移酶活性随处理时间的延长整体趋势表现出被诱导。     

Effect of stress and sympathetic activity on rat cardiac and aortic ornithine decarboxylase activity.

Adult male rats were injected either with α- or ß-adrenergic agonists and/ or antagonists and ornithine decarboxylase (ODC) activity in the heart and aorta was measured 4 hours later. At the lower doses, isoproterenol (0.2–0.4 mg/kg) resulted in a 10-fold increase in cardiac ODC activity but caused no significant change in aortic ODC activity. In contrast, phenylephrine (1 mg/kg) caused a 4-fold increase in aorta but no change in cardiac ODC activity levels. Phenoxybenzamine pretreatment completely abolished the PE-induced increase whereas no change was seen in ISop injected animals. Similarly, pretreatment with propranolol blocked the ISop induced response on ODC activity but had no effect on the increases observed after PE. These data suggest that the sympathetic regulation of ODC activity levels is mediated primarily via the ß-receptor in the heart but through the α-receptor in the aorta.     

Antibacterial and proteolytic activity of rabbit phagocytes and the effect of temperature and endotoxin on this activity

Proteolytic activity at pH 2 (caused by cathepsin E) and at pH 3.5 (caused by cathepsin D and E) was found in extracts of rabbit polymorphonuclear leucocytes, macrophages and lymphocytes, while antibacterial activity was detected only in polymorphonuclear leucocyte extracts, against microorganisms of theEnterobacteriaceae family. Comparison of the character of antibacterial and proteolytic activity showed that the two systems were quite different. If polymorphonuclear leucocyte extract was heated at 56° C for 30 minutes, the effect on antibacterial activity was non-significant, while proteolytic activity at pH 2 and 3.5 fell by about 60%. The addition of a heat-inactivatedEscherichia coli suspension or of purifiedSalmonella paratyphi B bacterial lipopolysaccahride to leucocyte extract completely destroyed antibacterial activity, while proteolytic activity was unaffected.     

PLTP activity inversely correlates with CAAD: effects of PON1 enzyme activity and genetic variants on PLTP activity

Recent studies have failed to demonstrate a causal cardioprotective effect of HDL cholesterol levels, shifting focus to the functional aspects of HDL. Phospholipid transfer protein (PLTP) is an HDL-associated protein involved in reverse cholesterol transport. This study sought to determine the genetic and nongenetic predictors of plasma PLTP activity (PLTPa), and separately, to determine whether PLTPa predicted carotid artery disease (CAAD). PLTPa was measured in 1,115 European ancestry participants from a case-control study of CAAD. A multivariate logistic regression model was used to elucidate the relationship between PLTPa and CAAD. Separately, a stepwise linear regression determined the nongenetic clinical and laboratory characteristics that best predicted PLTPa. A final stepwise regression considering both nongenetic and genetic variables identified the combination of covariates that explained maximal PLTPa variance. PLTPa was significantly associated with CAAD (7.90 × 10⁻⁹), with a 9% decrease in odds of CAAD per 1 unit increase in PLTPa (odds ratio = 0.91). Triglyceride levels (P = 0.0042), diabetes (P = 7.28 × 10⁻⁵), paraoxonase 1 (PON1) activity (P = 0.019), statin use (P = 0.026), PLTP SNP rs4810479 (P = 6.38 × 10⁻⁷), and PCIF1 SNP rs181914932 (P = 0.041) were all significantly associated with PLTPa. PLTPa is significantly inversely correlated with CAAD. Furthermore, we report a novel association between PLTPa and PON1 activity, a known predictor of CAAD.     

Influence of water activity on the ice-nucleating activity of Pseudomonas syringae

Pseudomonas syringae is known as a biological ice-nucleating agent. The bacterium has the unusual property of increasing the temperature at which water freezes by a few degrees. However, the ice-nucleating activity (INA) always remains lower for in vitro cultivated cells, than for cells grown in planta. We examined the effects of the hydrophobic environment and of water availability, on the in vitro growth and INA of P. syringae. The hydrophobic environment was modified by addition of fatty acids, vegetable oils or silicone oil to the culture medium. Addition of olive oil (1%), or traces of silicone oil in the culture medium had a positive effect upon the expression of INA. Variations in water activity from 0.990 to 0.988 by addition of sugar beet fibres or sodium chloride in the culture medium were followed by an increase in INA. This study suggested that control of the medium’s water activity must be considered as an important parameter for optimization of INA in P. syringae. Received 16 June 1998/ Accepted in revised form 02 September 1999     

Electromyography activity across gait and incline: The impact of muscular activity on human morphology

The study of human evolution depends upon a fair assessment of the ability of hominin individuals to gain access to necessary resources. We expect that the morphology of extant and extinct populations represents a successful locomotory system that allowed individuals to move across the environment gaining access to food, water, and mates while still maintaining excess energy to allocate to reproduction. Our assessment of locomotor morphology must then incorporate tests of fitness within realistic environments—environments that themselves vary in terrain and whose negotiation requires a variety of gait and speeds. This study assesses muscular activity (measured as the integrated signal from surface electromyography) of seven thigh and hip muscle groups during walking and running across a wide range of speeds and inclines to systematically assess the role that morphology can play in minimizing muscular activity and thus energy expenditure. Our data suggest that humans are better adapted to walking than running at any slope, as evidenced by small confidence intervals and even trends across speed and incline. We find that while increasing task intensity unsurprisingly increases muscular activity in the lower limb, individuals with longer limbs show significantly reduced activity during both walking and running, especially in the hip adductors, gluteus maximus, and hamstring muscles. People with a broader pelvis show significantly reduced activity in the hip adductor and hamstring muscles while walking. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

Effects of repeated blood samplings on locomotor activity, evasion and wheel-running activity in mice

The effects of serial blood sampling on nocturnal locomotor activity, evasion, wheel-running activity and body mass were studied in male NMRI mice aged 7-8 weeks. The erythrocyte count, haematocrit and haemoglobin concentration at the beginning and end of the study showed no difference in group 1 (two samples per week, 0.08 ml each) while there was a significant decrease in the group 2 values (three samples per week, 0.08 ml each). The total amount of nocturnal locomotor activity decreased in the animals bled repeatedly while the periods with locomotor activity increased. These alterations appeared particularly after bleeding. In the test-group animals evasion showed a decrease compared with the untreated control animals, but there was no evidence of a relation to the timing of the bleedings.     

Effects of weight reduction on neutrophil phagocytic activity and oxidative burst activity in female judoists

This study investigated the effects of short‐term weight reduction on neutrophil functions in female judoists. Sixteen actively competing female judoists were divided into two groups. Eight who required weight reduction were de?ned as the weight reduction group, and the remaining eight were used as the control. Blood samples were taken before, immediately after and 8 days after the match. Phagocytic activity and oxidative burst activity of neutrophils were measured by ?ow cytometry. In the weight reduction group, the phagocytic activity per cell decreased signi?cantly at the end of weight reduction compared with the control group. The rate of neutrophils producing reactive oxygen species and the oxidative burst activity per cell increased signi?cantly at the end of weight reduction in both the control and the weight reduction groups. We concluded that weight reduction, consisting of both intense exercise and energy restriction, might possibly cause both an increase in oxidative burst activity and decrease in neutrophil phagocytic activity in female judoists. However, although exercise increased oxidative burst activity, it did not affect neutrophil phagocytic activity alone. Therefore, to avoid this problem, female judoists are recommended to keep their weight within those limits determined by their class, and which can be reduced by exercise. Copyright © 2003 John Wiley & Sons, Ltd.     

Bacteriocin activity and probiotic activity of Aeromonas media

Three strains of Aeromonas media (161, A164 and A199) were shown to be active in-vitro producers of bacteriocin-like inhibitory substances (BLIS). For example, the producer strain, Aer. media A199, displayed antagonistic activity against all strains tested of Aer. caviae, Aer. hydrophila, Aer. salmonicida, Aer. veronii var. sobria, Listonella anguillarum, Photobacterium damsella, eight species of Vibrio and Yersinia ruckeri. Because of this wide-ranging activity against fish/shellfish pathogens, A199 was chosen for the probiotic work. By contrast, however, the BLIS produced by A199 did not inhibit the growth of Enterococcus seriolicida. The aim of the project was to ascertain whether or not the activity observed in vitro could be repeated in vivo. The ability of BLIS-producing strain A199 to act as a probiotic was assessed on the host animal, Crassostrea gigas, by testing whether or not strain A199 could prevent death of the oyster larvae when challenged with V. tubiashit. Whereas larvae, challenged with the Vibrio, died within 5 days, the presence of both the pathogen and the probiotic strain, together, did not affect the viability of the larvae over the same time period; the viability of larvae challenged with A199 alone was also unaffected when compared with the viability of unchallenged larvae (controls). These findings have important, economic implications for those engaged in the oyster producing industry where heavy losses can be experienced as a result of an infectious outbreak. At this stage, the association between BLIS activity and probiotic activity is circumstantial and, hence, future work will involve the use of non-BLIS-producing strains of Aer. media and BLIS-negative variants of the producer. Moreover, extension of the project will involve the use of other BLIS-producing strains (A161, and A164), hosts (salmon, crayfish, scallops and abalone) and pathogens.     

Reconstitution of heme-synthesizing activity from ferric ion and porphyrins, and the effect of lead on the activity

We examined the activity of heme synthesis when ferrochelatase purified from rat liver mitochondria was incubated with ferric chloride and mesoporphyrin IX as substrates in the absence of reducing reagents. In the presence of the NADH dehydrogenase-rich fraction and NAD(P)H, mesoheme was synthesized; the addition of FMN or FAD markedly enhanced the activity. These results indicate that the NAD(P) H-oxidizing system reduces ferric ion to ferrous ion. This ferrous ion is then utilized for heme synthesis by ferrochelatase. The effect of lead on NAD(P)H-dependent heme synthesis was also examined. Lead reduced NAD(P)H-dependent heme synthesis by 50% at 10(-5) M, but had no effect when ferrous ion was used as substrate. Zn-Porphyrin synthesis was not changed in the presence of Pb2+ at 10(-5) M. Thus, heme synthesis from ferric ion was more susceptible to Pb2+ than heme synthesis from ferrous ion.