Extraction of Pumpkin Polysaccharide by Complex Enzyme Method and Its Antioxidant Research

[Objective] This paper aims to study a new method of extracting pumpkin polysaccharide from pumpkin. Single factor experiments were conducted to examine the effects of extracting time,temperature,the solid-liquid ratio and pH value on the extraction yield of polysaccharide from pumpkin. [Method] The best enzyme ratio and extraction conditions for complex enzymes extraction were determined through orthogonal tests. Scavenging ·OH and O-2 activities of pumpkin polysaccharides were also investigated by salicylic acid and improved self-oxidation of o-pheno methods respectively. [Results] The results showed that the biggest extraction yield of polysaccharide from pumpkin can be got when adding 1% cellulose enzyme,1.5% pectinase,1.0% papain and Na2HPO4-citric acid buffer solution (pH was 4.6),and oscillating for 30 min under water at 40 ℃ with the solid-liquid ratio of 1:30. In addition,pumpkin polysaccharides had a strong activity of eliminating ·OH,but very weak activity to scavenge O-2. [Conclusion] This study provided basic data for research and application of Pumpkin polysaccharide.

Effects of Non-starch Polysaccharide Complex Enzymes on Meat Quality in Broilers

[Objective] The study aimed to research the effect of several commercial NSP complex enzymes products on broiler meat quality, and provide scientific basis for feed enterprise and breeding farmers choosing NSP complex enzymes. [ Method] Two hundred ROSS broilers at age of 21 days were designed to five treatment groups, including the positive and negative control treatments, and Group 1 to 3 with feed additive of the commercial non-starch polysaccharide complex enzymes each on base of the negative group diet. At 56 days of age, broilers were killed and meat quality was analyzed. [ Result] The thigh meat color CIELAB a value for the negative control broilers was lower （ P 〈 0.05） than those of the positive control and Group 1. The drip loss of breast meat for Group 1 was the lowest, and the drip loss of thigh meat for the negative control was the highest among all treatments. The shear force for the negative control and Group 3 were higher than those of other three groups. There were no differences （P 〉 0.05） on the contents of chemical components, crude moisture, crude protein, crude fat, crude ash, and inosine acid for breast and thigh meat among all treatments. [ Conclusion] NSP complex enzymes with complete enzyme categories and high enzyme activity can improve meat quality in broilers.

Study on Process Conditions of Preparation of Microporous Potatoes Starch by Complex Enzyme Method

[Objective] The technology of using c-amylase and glucoamylase to prepare microporeus potato starch was studied.[ Method ] Taking potato starch as raw materials, starch hydrolysis rate and the oil absorption as a measure of index, the influences of the reaction temperature, two enzymes proportion, the quantity of enzyme, the chroma of substrate, buffer solution pH and reaction time on microporous potato starch were inves- tigated. [ Result] The experimental results showed that the best technological conditions were reaction temperature 45 ℃, enzyme ratio （ glucoamy- lase/α-amylase）6, the quantity of enzyme （amount of enzyme and starch quality than） is 1.0%, the substrate quantity chroma of 0.14 g/ml, buffer solution pH 4, the reaction time 8 h. In such process condition, the oil adsorption rate of hydrolyzed potato starch was as high as 70.2%, starch hydrolytic ratio was 34.16%. [ Condmion] The study provided a basis for the development and utilization of microporous starch. Key words Microporous starch; Hydrolysis rate; Oil absorption rate; Preparation; Complex enzyme method; China

A Fiber Optic Sensor for the Simultaneous Measurement of Dual-parameter Based on Hydrogel-immobilized Enzyme Complex

A novel fiber optic sensor based on hydrogel-immobilized enzyme complex was developed for the simultaneous measurement of dual-parameter,the leap from a single parameter detecting fiber optic sensor to a fiber optic sensor that can continuously detect two kinds of parameters was achieved.By controlling the temperature from high to low,the function of fiber sulfide sensor and fiber DCP sensor can be realized,so as to realize the continuous detection of dual-parameter.The different variables affecting the sensor performance were evaluated and optimized.Under the optimal conditions,the response curves,linear detection ranges,detection limits and response times of the dual-parameter sensor for testing sulfide and DCP were obtained,respectively.The sensor displays high selectivity,good repeatability and stability,which have good potentials in analyzing sulfide and DCP concentration of practical water samples.

Electrophoretic Purification and Characterization of Human NADH-Glutamate Dehydrogenase Redox Cycle Isoenzymes Synthesizing Nongenetic Code-Based RNA Enzyme

NADH-glutamate dehydrogenase (GDH) is active in human tissues, and is chromatographically purified, and studied because it participates in synthesizing glutamate, a neurotransmitter. But chromatography dissociates the GDH isoenzymes that synthesize nongenetic code-based RNA enzymes degrading superfluous mRNAs thereby aligning the cellular reactions with the environment of the organism. The aim was to electrophoretically purify human hexameric GDH isoenzymes and to characterize their RNA enzyme synthetic activity as in plants. The outcome could be innovative in chemical dependency diagnosis and management. Multi metrix electrophoresis including free solution isoelectric focusing, and through polyacrylamide and agarose gels were deployed to purify the redox cycle isoenzymes of laryngeal GDH, and to assay their RNA enzyme synthetic activities. The laryngeal GDH displayed the 28 binomial isoenzymes typical of higher organisms. Isoelectric focusing purification produced pure GDH. Redox cycle assays of the GDH isoenzymes produced RNA enzymes that degraded human stomach total RNA. In the reaction mechanism, the Schiff-base intermediate complex between α-ketoglutarate and GDH is the target of nucleophiles, resulting to the disruption of synthesis of glutamate, and RNA enzyme. The strongest nucleophiles are the psychoactive alkaloids of tobacco, cocaine, opium poppy, cannabis smoke because they are capable of reacting with GDH Schiff base intermediate to stimulate synthesis of aberrant RNA enzymes that degrade cohorts of mRNAs thereby changing the biochemical pathways and exacerbating drug overdose and chemical dependency. Electrophoretic purification, and characterization of the RNA enzyme synthetic activity set the forecourt for innovative application of GDH redox cycles in the diagnostic management of chemical dependency.

Mitochondrial energy metabolism changes during aging-mouse cranial nerve cells treated with various doses and forms of Fructus schizandrae

BACKGROUND： During the cellular aging process, the number of mitochondria, generation of adenosine triphosphate （ATP）, activity of respiratory chain enzyme complex 1 and 4, and oxidation decrease. OBJECTIVE： To observe the effects of aqueous and spirituous extract, as well as polysaccharides from Fructus schizandrae （Magnolia Vine） on energy metabolism and mitochondrial anti-oxidation in cranial nerve cells of a D-gal-induced aging mouse model. DESIGN, TIME AND SETTING： A randomized, controlled, animal study. The experiment was conducted at the Department of Biochemistry, Qiqihar Medical College between March and July 2006. MATERIALS： Fifty healthy, Kunming mice of both sexes, aged 2 3 months old and weighing 18 22 g, were used for the present study. Fructus schizandrae was purchased from the Medical College of Jiamusi University. Aqueous extracts, spirituous extracts, and polysaccharides from Fructus schizandrae were prepared. D-galactose （D-gal） is a product of the Second Reagent Factory, Shanghai City, China. Mn-superoxide dismutase （Mn-SOD） kit, malonaldehyde （MDA） kit, protein quantification kit, and inorganic phosphorus testing kit were purchased from Jian Cheng Bioeng. Co., China. METHODS： Fifty mice were randomly divided into five groups, with 10 mice in each group： young control, aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides. Over a course of 30 days, mice in aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides groups were injected subcutaneously with D-gal （100 mg/kg） into the nape of the neck daily, and administered intragastrically with an equal volume of sterile, warm water （aging model）, aqueous Fructus schizandrae extract （2 g/kg）, spirituous Fructus schizandrae extract （2 g/kg）, or Fructus schizandrae polysaccharides （0.2 g/kg）, respectively. Mice in the young control group were injected into the nape of the neck with physiological saline and administered intragastrically with sterile, warm water. MAIN OUTCOME MEASURES： Respiratory chain complex I and H^＋-ATP enzyme activities, as well as Mn-SOD and MDA levels, were determined by the Coomassie brilliant blue method. RESULTS： All fifty mice were included in the final analysis. In mitochondria fiom cranial nerve cells of the aging mouse group MDA levels were significantly increased, compared with young control group （P 〈 0.01）; however, Mn-SOD levels, as well as respiratory chain complex I and H＋-ATP enzyme activity, were remarkably decreased （P 〈 0.01 ）. In each Fructus schizandrae group, Mn-SOD levels, as well as respiratory chain complex I and H＋-ATP enzyme activity was enhanced to various extents （P 〈 0.05, P 〈 0.01）, and MDA levels were decreased （P 〈 0.01）, compared with the aging model group. CONCLUSION： Aqueous and spirituous Fructus schizandrae extracts, as well as Fructus schizandrae polysaccharides delayed changes in mitochondrial energy metabolism, increased Mn-SOD levels, and decreased MDA levels in cranial nerve cell mitochondria of an aging mouse model. Fructus schizandrae polysaccharides were particularly capable of protecting mitochondria from oxidative injury.

Improving pathway prediction accuracy of constraints-based metabolic network models by treating enzymes as microcompartments

Metabolic network models have become increasingly precise and accurate as the most widespread and practical digital representations of living cells.The prediction functions were significantly expanded by integrating cellular resources and abiotic constraints in recent years.However,if unreasonable modeling methods were adopted due to a lack of consideration of biological knowledge,the conflicts between stoichiometric and other constraints,such as thermodynamic feasibility and enzyme resource availability,would lead to distorted predictions.In this work,we investigated a prediction anomaly of EcoETM,a constraints-based metabolic network model,and introduced the idea of enzyme compartmentalization into the analysis process.Through rational combination of reactions,we avoid the false prediction of pathway feasibility caused by the unrealistic assumption of free intermediate metabolites.This allowed us to correct the pathway structures of L-serine and L-tryptophan.A specific analysis explains the application method of the EcoETM-like model and demonstrates its potential and value in correcting the prediction results in pathway structure by resolving the conflict between different constraints and incorporating the evolved roles of enzymes as reaction compartments.Notably,this work also reveals the trade-off between product yield and thermodynamic feasibility.Our work is of great value for the structural improvement of constraints-based models.