CHARACTERIZATION OF PREDICTED BACTERIAL COLD-ADAPTED LIPASE FROM SEAFOOD COLD STORAGE

Lipases constitute as top three most important group of enzymes along with carbohydrases and proteases, and are widely used in various industries. In particular, lipase that perform high activity at low temperatures, or referred as cold adapted lipase (CLPs) considered as attractive catalyst due to its activity at low temperature. This unique feature is the main advantage of cold adapted lipase utilization because it requires a low energy source that is correlated with lower production costs and energy. In addition, reactions occur in cold temperatures may result in better product quality. The purpose of this research was to perform screening and characterization of bacterial cold adapted lipase from seafood cold storage. Among 53 isolates, Kr_16_30, TI_37_14 and Kr_16_28 showed the highest activity with 4.12 U/mL; 3.87 U/mL and 3.21 U/mL, respectively. Isolates Kr_16_30 seemed to be typical cold adapted lipase with optimum temperature at 20°C and pH 7. Isolates Kr_16_28 performed highest lipolytic activity at 30°C while TI_37_14 suspected to be similar to typical mesophilic lipase with optimum temperature at 40°C. Species identification based on 16s rRDA sequencing revealed that isolates Kr_16 30 and Kr_16 28 are belong to genus Pseudomonas and Bacillus, repectively.

Investigation of lipolytic activity of the red king crab hepatopancreas homogenate by NMR spectroscopy

The digestive gland of craboids (hepatopancreas) is rich in a huge number of various enzymes (collagenases, nucleases, hyaluronidases, proteases), which are well studied at the moment. However, little is known about crustacean lipases. In this work, using 1H NMR spectroscopy, it was found that the hepatopancreas homogenate of the red king crab Paralithodes camtschaticus demonstrates high lipolytic activity against triacetin in a wide pH range and shows moderate activity against the caprylic/capric triglyceride emulsion. Under the action of the hepatopancreas homogenate, triacylglycerols are converted into 1,2-diacylglycerol, and then into 2-monoacylglycerol and 1-monoacylglycerol. The 1-monoacylglycerol predominates in the reaction products. The use of NMR spectroscopy makes it possible to quickly detect hydrolysis products and evaluate the reaction direction.

Macronutrients and essential amino acids on digestive process of the freshwater teleost Matrinxã

The objective of this work was to evaluate the effect of macronutrients and essential amino acids on digestive process of the freshwater teleost Matrinxã (Brycon amazonicus). Juveniles were fed with diets containing starch plus free amino acids or oil plus free amino acids for 15 days. These fish were compared with others fed with diets containing starch or oil without addition of free amino acids. After the experimental span, 12 fish from each treatment were randomly sampled to collect stomach, pyloric cecum, anterior and posterior intestine for assaying digestive enzymes activity. Increase of gastric proteolysis due to dietary amino acids were observed. Amylolytic, proteolytic and lipolytic activities in intestine sections were also positive related to dietary amino acids. However, proteolytic and lipolytic activities in pyloric cecum were not responsive to dietary changes. Moreover, the absence of starch in the diets resulted in decrease of amylolysis, and very low levels of oil did not change the lipolytic activity. In conclusion, activities of amylase, protease and lipase of Matrinxãare selectively responsive to addition of free essential amino acids concerning the gut section.

Powerful Lipolytic Activity of Fungi Isolated from Coconut and Avocado Flesh on Different pH and Temperature

Lipase is widely applied in various fields of industries elevating its economic value. The demand for lipase keeps increasing open up working opportunities. Isolates showing lipolytic activity obtained previously are tested for their activity on different pH mediums: 7, 8, 9, and 10; and at different temperatures. The fungi are grown on SDA medium supplemented with Olive Oil, emulsifier, Tween 80, and Rhodamine B. The experiment was made in 3 replicates and is incubated at room temperature of 30oC. Lipase activity was calculated based on the clear zone around colonies observed on day 2 and day 3 after inoculation. The result shows that those isolates are highly active on various pH at RT, and the activity slightly reduces at 30oC. This result suggests that a wide range of their applications are at room temperature when pH is a limiting factor for the applications

Partial purification, characterization and immobilization of a novel lipase from a native isolate of Lactobacillus fermentum

Background and Objectives: Due to the widespread use of lipase enzymes in various industries, finding native lipase pro- ducing microorganisms is of great value and importance. In this study, screening of lipase-producing lactobacilli from native dairy products was performed. Materials and Methods: Qualitative evaluation of lipolytic activity of lipase-producing lactobacilli was performed in differ- ent media containing olive oil. A clear zone observation around the colonies indicated the lipolytic activity. The strain with the highest enzymatic activity was identified. Determination of optimal pH and temperature of lipase activity was measured by spectrophotometry using p-nitrophenyl acetate (ρ-NPA) substrate. Partial purification of lipase enzyme was performed using 20-90% saturation ammonium sulfate. Eventually, lipase was immobilized by physical adsorption on chitosan beads. Results: Among screened lipolytic bacterial strains, one sample (5c isolate) which showed the highest enzymatic activity (5329.18 U/ml) was close to Lactobacillus fermentum. During characterization, the enzyme showed maximum activity in Tris-HCl buffer with pH 7, while remaining active over a temperature range of 5°C to 40°C. The results of the quantitative assay demonstrated that the fraction precipitated in ammonium sulfate at 20% saturation has the highest amount of lipolytic activity, with a specific activity of 22.0425 ± 3.6 U/mg. Purification folds and yields were calculated as 8.73 and 44%, respec- tively. Eventually, the enzyme was immobilized by physical adsorption on chitosan beads with a yield of 56.21%. Conclusion: The high efficiency of enzyme immobilization on chitosan beads indicates the suitability of this method for long-term storage of new lipase from native 5c isolate.

Digestive Activity of Pancha Harithakadi Churna

The traditional systems of medicine are really effective but the problem with them is they lack in quality assurance. Standardization is the need of the hour in ayurvedic system of medicine. PanchaHarithakadi Churna (PHC) is a traditional polyherbal formulation which consists of five household ingredients used for indigestion. It is mainly used for Constipation and Bloating. Churna’s will play a major role in gastro intestinal problems and they have greater bioavailability because of smaller particle size. It consists of fine powder (sieve 100 size) of ginger rhizomes, fennel fruits, myrobalan fruits, senna leaflets and pink rock salt in equal proportions (1:1:1:1:1) are mixed well. PHC was formulated by standard procedures and evaluated by microscopic characterization, inorganic analysis and digestive studies. Microscopical characters indicate the presence of genuine crude drugs used in the formulation. Inorganic analysis shows the presence of calcium, magnesium, sodium, chloride and phosphate. The PHC showed pronounced amylolytic activity and trypsin activity whereas moderate lipolytic activity, proteolytic activity and pepsin activity, mild chymotrypsin activity in treating indigestion. In future we will carry out in vivo digestive studies.

Technological Parameters, Anti-Listeria Activity, Biogenic Amines Formation and Degradation Ability of L. plantarum Strains Isolated from Sheep-Fermented Sausage

The aim of this work was to identify and characterize, from a technological and safety point of view, the lactic acid bacteria (LAB) isolated from traditional sheep-fermented sausage. First, LABs were identified then were screened for some technological parameters such as acidifying and growth ability, proteolytic and lipolytic activity and for antimicrobial activity. Finally, biogenic amine production and degradation abilities were also evaluated. This research reveals the predominance of Lactiplantibacillus (L.) plantarum on LAB community. Almost all L. plantarum strains were active against Listeria monocytogenes strains (inhibition zone diameters > 1 cm). None of the tested strains were positive in histidine (hdcA), lysine (ldc) and tyrosine (tyrdc) decarboxylase genes and only one (L. plantarum PT9-2) was positive to the agmatine deiminase (agdi) gene. Furthermore, given the positive results of the sufl (multi-copper oxidase) gene detection, all strains showed a potential degradation ability of biogenic amines.

Changes in lipase activity when using gelatinous agents

Липолитическая порча, приводящая к появлению мыльного привкуса, является браковочным признаком кондитерских изделий. Исследовано влияние студнеобразователей на активность липолитических ферментов, Растворы пектина и желатина не оказали влияния на липолитическую активность, а 1%-ный раствор агара увеличил ее на 10 %. Результаты работы способствуют повышению сохранности кондитерских изделий. Lipolytic spoilage, which leads to the appearance of a soapy taste, is a defective sign of confectionery products. The effect of gelatinous agents on the activity of lipolytic enzymes was studied, pectin and gelatin solutions did not affect the lipolytic activity, and 1 % agar solution increased by 10 %. The results of the work contribute to improving the safety of confectionery products.

Characterization of preparation for fat separators

Fats, oils and greases in waste water frequently cause serious environmental problems. A commercial preparation was tested for its ability to degrade oil and grease in wastewater. As substrates, two fats (lard and beef fat) and four oils (rape seed oil, sunflower oil, palm oil and olive oil) were chosen. The degradation ability of commercial preparation for different substrates was investigated by the determination of lipid degradation at pH 7, temperature 25 °C, aerobic condition and agitation at 160 rev/min for 14 days in Erlenmeyer flasks. Simultaneously, the lipolytic activity was spectrophotometrically determined at 420 nm. All tested substrates were degraded by the different rate to basic units (fatty acids). Rape seed oil and lard were decomposed by bacterial lipase from commercial preparation.

Effect of Resistance Exercise on the Lipolysis Pathway in Obese Pre- and Postmenopausal Women

Physical exercise may stimulate lipolytic activity within adipose tissue. Furthermore, resistance exercise may contribute to the more efficient reduction in adipose tissue mass and prevent the accumulation thereof in obese women. The purpose of this study was to examine the effects of regular resistance exercise for 12 weeks on the lipolysis pathway in women with obesity. Twenty-three pre- and postmenopausal women with body fat percentages of 30% or more were divided into the premenopausal group (n = 9) and the postmenopausal group (n = 14). All subjects participated in resistance exercise training for 12 weeks. Anthropometric and physical fitness tests were performed on all participants. Protein analyses were performed on extracted subcutaneous fatty tissue, and changes in the relevant protein levels in the samples were analyzed by Western blotting. All serum samples were submitted for enzyme-linked immunosorbent assay measurements of adipocyte factors. After 12 weeks, the adipose triglyceride lipase, monoacylglycerol lipase, and perilipin1 protein levels were significantly lower in the postmenopausal group than in the premenopausal group. The hormone-sensitive lipase protein levels were significantly higher in the postmenopausal group than in the premenopausal group. In addition, leptin concentrations were significantly decreased after resistance exercise in the postmenopausal group. Adiponectin concentrations were significantly increased after resistance exercise in both groups. These findings indicate that regular resistance exercise is effective in reducing the weight and body fat of obese premenopausal women, and in the secretion of adiponectin. On the other hand, postmenopausal women were found to have redeced weight and body fat, and were found to be positive for the secretion of adipokine factors. In addition, positive changes in lipolysis pathway factors in adipose tissue promote lipid degradation and reduce fat mass. Thus, regular resistance exercise shows positive changes in the lipolysis pathway more effectively in weight and body fat reduction in postmenopausal women than in premenopausal women.