As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.
The effects of amylase addition during extrusion cooking of rice on the physico-chemical properties of the extrudates were investigated in order to develop rice-based Korean style weaning food products. A laboratory-built single screw extruder was used, the enzymes used were Termamyl 120LS(amylase from Bacillus licheniformis, NOVO Co.), BAN 240L(amylase from Bacillus amylolichuefaciens, NOVO Co.) and malt powder. By the addition of enzymes, the water soluble index of the extrudates increased by 3 times compared to that of the extrudates without enzyme and the concentration of reducing sugar in the extrudates increased drastically at 28 feed moisture. The gel permeation chromatographic pattern showed that the large molecular starch fractions diminished by the addition of enzyme during extrusion and the low molecular fraction increased. The residual enzyme activity in the extrudate were 27% for the most thermo-resistance enzyme by treating at $140^{\circ}C$ in the metering section of the extruder. The sediment volume of the extrudate dispersion increased as the metering section temperature increased to $140^{\circ}C$ . By the addition of enzymes the viscosity of extrudate dispersion was redused $1/4{\sim}1/200$ of that of the extrudates without enzyme. It allowed to use 1.8 times of solid material to the weaning food formulation to attain the same level of consistency as the commercial products. It proves that the addition of amylase during rice extrusion is effective to increase the energy density of weaning food by starch degradation, which results in the increases of water solubility, reducing sugar content, dispersibility and fluidability.
Journal of the Korean Society of Food Science and Nutrition
/
v.31
no.4
/
pp.685-690
/
2002
The effects of mycelium of Cordyceps militaris on the growth, the lipid metabolism, the serum protein levels and the enzyme activities in male rats were studied. Sprague-Dawley rats were given four different types of diets for a succeeding period of five weeks: either a control diet, a control diet supplemented with 2%, 3% or 4% mycelium of Cordyceps militaris (CM) powder. The body weight gain, hepatic weight, feed efficiency ratio and the feed intake of the rats given diets with 2%, 3% or 4% CM were similar to those in rats given the control diet. The concentrations of hepatic total lipid and triglyceride of rats fed the 3% or 4% CM diets were significantly lower than those of rats fed the control diet. But the concentrations of hepatic total cholesterol and phospholipid of rats fed the all CM diets were similar to those of rats fed the control diet. The concentrations of total lipid, total cholesterol, triglyceride phospholipid, and the atherogenic index, and the activities of glutamic oxaloacetic transaminase and lactic dehydrogenase in serum of rats fed the all CM diets were significantly lower than those of rats fed the control diet. No differences were noted in the concentrations of HDL-cholesterol, LDL-cholesterol, total protein, albumin and creatinine, and the activities of glutamic pyruvic transaminase, ${\gamma}$ -glutamyltranspeptidase and alkaline phosphatase in the serum among the rats with on all the experimental diets. These results showed that the all CM diets feeding decreased the total cholesterol, the triglyceride, the phopholipid, and the atherogenic index in serum of rats.
Changes of silk protein compositions of average molecular weight (Mw) and free amino acid composition to different solubility conditions were studied by SDS-polyacrylamide electropholesis, gel permeation chromatography (GPC), and free amino acid analysis method. We can not detected average molecular weight distribution of different hydrochloric acid (HCl) conditions as SDS-polyacrylamide method, but as using GPC method, molecular weight distribution of 2N-HCl, 1N-HCl and 0.5N-HCl (3 hrs at $110^{\circ}C$ treated) are confirmed Mw 800, 1,500 and 3,700, respectively. The average molecular weight of calcium chroride and calcium chloride-enzyme treated samples are shown Mw 46,800 and 12,500, respectively. The degree of hydrolysis and the composition of the free amino acid in the fibroin hydrolysates effected significantly composition of free amino acids of the fibroin powder. The increase of the degree of hydrolysis and ratio of free amino acids and oligopeptides were found to be directly related to the concentration of hydrochloric acid and treatment of enzyme, resulting in the increase of water solubility.
A Trichoderma sp. SO-571 producing cellulase was isolated from soil, and a pilot-scale cultivation and separation of cellulase were conducted. The cellulase activity was about 14.5 unit/ml after 112 hr of cultivation in a 301 fermenter containing 3.0% cellulose, 4.0% soybean powder, 3.0% wheat bran, 0.5% ($NH_4$)$_2$$SO_4$0.2% urea, 1.0% CSL, 0.5% $KH_2$PO$_4$, and 0.2% Tween 80. The cellulase was purified over 4.6 folds in three steps with 47.86% yield. The optimum pH of cellulase was pH 5.0 and optimum temperature was $60^{\circ}C$. To investigate the effect of the cellulase-treated cellulosic fabric, the weight loss was compared. The weight loss of denim treated with cellulase from Trichoderma sp.SO-571 was 2.9% and that with Celluclast 1.5L was 2.2%. In tencel treatement with enzyme, cellulase showed 0.7% higer weight loss than that with Celluclast 1.5L.
The survey on the chitinolytic activity of some plants was performed for the purpose of obtaining some reliable and inexpensive sources of chitinase. Rice, soybean for sprouting, kiwi fruit, almond and crude papain were investigated. Rice bran, seed coat of the soybean and the pericarp of kiwi fruit showed a considerable activity, while the bean after the removal of the seed coat, the mixture of rice integument and endosperm, polished rice, and defatted soybean powder didn't have any detectable activity. These crude enzymes have shown to contain both endo- and exochitinase activity. The effects of pH and temperature on the enzyme activity were variable. Furthermore we have observed the chitosanolytic activity from these enzyme Preparations. The rice bran had the highest activity in the enzymatic degradation of chitosan, and seed coat of soybean and the pericarp of kiwi fruit followed. On the basis of the fact that crude papain was not only commercially available but also the most potent in the endochitinase activity and the lowest in the exochitinase activity, we could conclude that crude papain was considered as the most suitable source of the chitinase among plants studied in this paper. In addition, rice bran was worth further investigation from the point of utilizing agricultural by-product.
Chae Won Lee;June Hyeok Yoon;Su Hyun An;In Ho Cho;Changsu Kong
Korean Journal of Poultry Science
/
v.50
no.3
/
pp.161-169
/
2023
The present study aimed to investigate the effect of Allium hookeri (AH) root on growth performance, serum antioxidant enzyme activity, cecal short-chain fatty acid profiles, and tight junction-related gene expression of broiler chickens. On day 10, 96 birds were allocated to one of two dietary treatments with 6 replicate cages, and 8 birds per cage in a randomized complete block design. The experimental diets consisted of a corn-soybean meal-based control diet and a diet supplemented with 0.3% AH root powder at the expense of the corn in the control diet. The experimental diets were formulated to meet or exceed the nutrients requirements of broilers recommended by Aviagen for each phase (from day 10 to 21 and from day 21 to 28). On day 28, a bird representing the median body weight in each cage was selected, and then blood, cecal digesta, and jejunum tissue samples were collected. No significant differences were observed in body weight gain and feed intake among dietary treatments. However, birds fed the diet containing AH showed a greater gain-to-feed ratio compared to the control group at 21 days of age and throughout the entire experimental period (P<0.05). Serum antioxidant enzyme activity, cecal short-chain fatty acid profiles, and tight junction-related gene expression were not affected by the dietary AH supplementation. Based on the results of the present study, dietary AH supplementation may improve the gain-to-feed ratio, but not gut health markers in broiler chickens from day 10 to 28.
Two experiments, involving a total of 100 crossbred pigs (Hampshire$\times$Landrace$\times$Duroc) aged 5 weeks, were used to evaluate the effect of Roselle (Hibicus sabdariffa Linn.) calyx as an antioxidant and acidifier on growth performance. Experiment 1: growth performance response of pigs fed basal corn-soy diet was compared with that of pigs consuming diets that contained 4, 8 and 12% Roselle in powder form, 4 g/kg acidifier (Fra$^{(R)}$Acid Dry) or 100 mg/kg antibiotic (Aurofac$^{(R)}$). All diets were isocaloric and isonitrogenous. Twenty-four castrated male and twelve female pigs were randomly allocated into 6 treatments which comprised 3 replicates (2 male and 1 female) of 2 pigs each. The results demonstrated no beneficial weight gain and feed intake response among treatments at 7, 9 and 11 weeks of age (p>0.05). However, feed-to-gain ratios (FCR) of Roselle-fed groups were significantly different from the basal-diet group at 7 weeks of age (p<0.05). Pigs fed with 8% Roselle had the lowest FCR. Therefore, 8% was set as an appropriate level of Roselle in feed. Experiment 2 was conducted to determine antioxidant and acidifier properties. Pigs were fed similar diets to Experiment 1 except that Roselle was fed to only one treatment at the level of 8%. Sixty-four piglets, 32 castrated male and female each were randomly allocated into 4 treatments which comprised 4 replicates (2 male and 2 female) of 4 pigs each. Body weight was measured. At 7 and 9 weeks of age, four pigs from each treatment were randomly selected. The pH in their gastrointestinal tract, pepsin activity in stomach mucosa, trypsin activity in the pancreas and protein and fat digestibility in the ileum were measured to investigate acidifier properties. For antioxidant properties, malondialdehyde (MDA) and glutathione concentration in plasma and liver were used as parameters. The results showed no significant difference in weight gain, pH or pepsin activity among all treatments at both ages (p>0.05). Nevertheless, trypsin activity and fat digestibility of the Roselle-fed group were higher than the basal-diet group at 7 weeks of age (p<0.05). Glutathione in plasma was significantly different between antibiotic-fed and basal-diet groups (p<0.05). Significant differences were found only in some parameters and between the basal-diet group and the other groups. Therefore, it cannot be precisely concluded that Roselle in powder form has clear acidifier and antioxidant properties in postweaning pigs. Further studies should be undertaken to clearly confirm both properties.
Kim, Kyeong-Yeol;Nam, Min-Ji;Nam, Bo-Ram;Ryu, Hee-Jung;Song, Jeong-Eon;Shim, Won-Bo;Lee, Soo-Hyung;Chung, Duck-Hwa
Journal of Environmental Health Sciences
/
v.36
no.1
/
pp.52-60
/
2010
High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.
Kim, Young-Joo;Seo, Sheungwoo;Wang, Xiaoyu;Seo, Dong Joo;Lee, Min Hwa;Son, Na Ry;Lee, Bog-Hieu;Choi, Changsun
Food Science of Animal Resources
/
v.33
no.5
/
pp.610-616
/
2013
Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplification of DNA under isothermal conditions. The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder. In order to enhance the sensitivity and specificity, LAMP primers targeting outer membrane protein A (ompA) gene of C. sakazakii were designed using Explorer V4 software. Thirty seven C. sakazakii strains and 13 pathogenic microorganisms were used for comparative detection of C. sakazakii using polymerase chain reaction (PCR), real-time PCR, and LAMP. LAMP developed in this study could specifically detect C. sakazakii strains without cross-reactivity with other foodborne pathogens. LAMP products amplified from ompA gene of C. sakazakii were digested with with HhaI and NruI enzyme. The specificity of LAMP was confirmed by restriction fragment length polymorphism (RFLP) analysis. LAMP could detect C. sakazakii within 1 h without bacterial culture and its detection limit was as low as 1 CFU/mL C. sakazakii in milk. In the comparison of the sensitivity, LAMP showed 10,000- and 100-times higher detection limit than PCR or real-time PCR, respectively. Therefore, this study can conclude that LAMP is a rapid and reliable detection technique for C. sakazakii contaminated in powdered milk.
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