Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays

2011 ◽  
Vol 414 (1) ◽  
pp. 169-171 ◽  
Author(s):  
Qingwen Chen ◽  
Kumaran Narayanan
Author(s):  
Yizhao Shen ◽  
Ranithri Abeynayake ◽  
Xin Sun ◽  
Tao Ran ◽  
Jianguo Li ◽  
...  

Abstract Background This study was conducted to evaluate the feed nutritional value of brewers’ spent grain (BSG) residue resulting from protease aided protein removal. The nutritional value was measured as nutrient content, gas production, nutrient digestibility and fermentation characteristics in batch culture. Results Protein extraction process decreased content of crude protein but concentrated the neutral detergent fibre (NDF) and ferulic acid in BSG residue. The changes in the chemical composition of BSG residue varied with enzyme and enzyme dosage. Digestibility of dry matter (DMD) and NDF of residue differed among proteases. Increasing alcalase dosage linearly decreased DMD, whereas, the DMD linearly increased as everlase or flavourzyme dosage increased. Compared with BSG, the DMD, gas production and fermentation acid concentration of BSG residues were lower, whereas NDF digestibility was higher. Conclusions The substantially increased NDF content and improved in vitro NDF digestibility due to protease hydrolysis suggest that BSG residue can be potentially exploited as a viable fibre source for ruminant feeding.


1983 ◽  
Vol 55 (2) ◽  
pp. 155-162
Author(s):  
Matti Näsi

The extraction of protein from leaves of two energy willow clones (Salic cv. Aquatica and S. viminalis) and the chemical composition and nutritive value of the leaf fractions were investigated in six experiments. The preservation of leaf juice and wet leaf protein concentrate (LPC) was also studied. The extraction of juice averaged 33 % of the fresh weight, and the extraction ratios of dry matter (DM) and crude protein (CP) were 10.5 % and 2.9 %. The percentages of juice DM, CP and TP (true protein) separated in LPC were 6 %, 21 % and 48 %, respectively. The separation values were higher when LPC was precipitated by heating than by acidification. Removal of nutrients from the leaves was low and there were only minor differences in composition between the pressed pulp and the leaves. The leaf juice contained 8.3 % DM, of which 20.9 % was ash, 4.9 % CP, 1.9 % TP and 43.2 % sugars. The LPC had an average DM content of 19.6 %, CP being 21.4 % and TP 19.5 % of DM. The in vitro digestibility and pepsin-HCI-solubility of the protein were low. The mean tannin content of the LPC averaged 3.6 % of DM. The mean lysine and methionine contents of the LPC were low, 3.3 and 1.5 g/16 g N, respectively. The preservation of the leaf juice was satisfactory even without addition of preservative. Preservation of wet LPC was good with additives used at the 1.0 % v/w level. Leaf protein extraction was low and the nutritive value of LPC was not particularly high. This may have been partly due to the late date of harvesting and to the fact that the leaves were frozen for storage.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yu-Cheng Chang ◽  
Yi-Ching Chiu ◽  
Nai-Wen Tsao ◽  
Yuan-Lin Chou ◽  
Choon-Meng Tan ◽  
...  

AbstractAmaranthus tricolor L., a vegetable Amaranthus species, is an economically important crop containing large amounts of betalains. Betalains are natural antioxidants and can be classified into betacyanins and betaxanthins, with red and yellow colors, respectively. A. tricolor cultivars with varying betalain contents, leading to striking red to green coloration, have been commercially produced. However, the molecular differences underlying betalain biosynthesis in various cultivars of A. tricolor remain largely unknown. In this study, A. tricolor cultivars with different colors were chosen for comparative transcriptome analysis. The elevated expression of AmCYP76AD1 in a red-leaf cultivar of A. tricolor was proposed to play a key role in producing red betalain pigments. The functions of AmCYP76AD1, AmDODAα1, AmDODAα2, and AmcDOPA5GT were also characterized through the heterologous engineering of betalain pigments in Nicotiana benthamiana. Moreover, high and low L-DOPA 4,5-dioxygenase activities of AmDODAα1 and AmDODAα2, respectively, were confirmed through in vitro enzymatic assays. Thus, comparative transcriptome analysis combined with functional and enzymatic studies allowed the construction of a core betalain biosynthesis pathway of A. tricolor. These results not only provide novel insights into betalain biosynthesis and evolution in A. tricolor but also provide a basal framework for examining genes related to betalain biosynthesis among different species of Amaranthaceae.


Animals ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 18
Author(s):  
Kim Margarette C. Nogoy ◽  
Jia Yu ◽  
Young Gyu Song ◽  
Shida Li ◽  
Jong-Wook Chung ◽  
...  

The amaranth plants showed high potential feed value as forage for ruminants. An in-depth study of this plant, particularly in cattle, will help extend its utilization as an alternative protein and fiber feed source in cattle feeding. In this study, the nutrient compositions of three different species of amaranth, Amaranthus caudatus L., Amaranthus cruentus L., and Amaranthus hypochondriacus L.—two varieties for each species, A.ca 74, A.ca 91, A.cu 62, A.cu 66, A. hy 30, and A. hy 48—were evaluated. The in vitro technique was used to evaluate the fermentation characteristics such as total gas production, total volatile fatty acids (VFA) concentration, pH, and ammonia concentration of the rumen fluid. Moreover, the effective degradabilities of dry matter (EDDM) and crude protein (EDCP) of the amaranth forages were determined through in situ bag technique. The amaranth forages: A. caudatus, A. cruentus, and A. hypochondriacus showed better nutritive value than the locally produced forages in Chungcheong province of Korea. The CP of the amaranth ranged from 11.95% to 14.19%, and the neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents ranged from 45.53% to 70.88% and 34.17% to 49.83%, respectively. Among the amaranth varieties, A. hypochondriacus 48 showed the most excellent ruminant feed nutrient quality (CP, 14.19%; NDF, 45.53%; and ADF, 34.17%). The effective degradabilities of dry matter (EDDM; 33–56%) and crude protein EDCP (27–59%) of the amaranth were lower compared to other studies, which could be due to the maturity stage at which the forages were harvested. Nonetheless, A. hypochondriacus 48 showed the highest EDDM (56.73%) and EDCP (59.09%). The different amaranth species did not differ greatly in terms of total VFA concentration or molar proportions, total gas production, or ammonia-N concentration. The high nutrient composition, and highly effective degradability of dry matter and crude protein, coupled with the favorable fermentation characteristics, suggest that the amaranth forages showed good to excellent feed quality for cattle.


2021 ◽  
Vol 22 (9) ◽  
pp. 4655
Author(s):  
Priyanka Bapat ◽  
Debalina Goswami Sewell ◽  
Mallory Boylan ◽  
Arun K. Sharma ◽  
Julian E. Spallholz

Her/2+ breast cancer accounts for ~25% mortality in women and overexpression of Her/2 leads to cell growth and tumor progression. Trastuzumab (Tz) with Taxane is the preferred treatment for Her/2+ patients. However, Tz responsive patients often develop resistance to Tz treatment. Herein, redox selenides (RSe-) were covalently linked to Tz using a selenium (Se)-modified Bolton–Hunter Reagent forming Seleno-Trastuzumab (Se-Tz; ~25 µgSe/mg). Se-Tz was compared to Tz and sodium selenite to assess the viability of JIMT-1 and BT-474 cells. Comparative cell viability was examined by microscopy and assessed by fluorometric/enzymatic assays. Se-Tz and selenite redox cycle producing superoxide (O2•−) are more cytotoxic to Tz resistant JIMT-1 and Tz sensitive BT-474 cells than Tz. The results of conjugating redox selenides to Tz suggest a wider application of this technology to other antibodies and targeting molecules.


1966 ◽  
Vol 66 (3) ◽  
pp. 351-357 ◽  
Author(s):  
W. Ellis Davies ◽  
G. ap Griffith ◽  
A. Ellington

The primary growth of eight varieties of three species–white clover (3), red clover (4) and lucerne (1)–was sampled at fortnightly intervals and the percentage dry matter, in vitro digestibility, crude protein, water soluble carbohydrates, P, Ca, K, Na and Mg were determined.Differences between species were nearly always significant and the general order of merit was white clover, red clover and lucerne. The exceptions were for dry-matter percentage where this order was reversed, and red clover had the lowest Na and highest Mg content.


2017 ◽  
Vol 39 (2) ◽  
pp. 137
Author(s):  
Marielly Maria Almeida Moura ◽  
Daniel Ananias de Assis Pires ◽  
Renê Ferreira Costa ◽  
Daniella Cangussú Tolentino ◽  
João Paulo Sampaio Rigueira ◽  
...  

 This study aimed to evaluate the nutritional characteristics and quality of silages of five sorghum genotypes, namely: Volumax, AG2005E, Qualimax, BRS610 and AG2501. The study was conducted at the Experimental Farm Unimontes, in the municipality of Janaúba, state of Minas Gerais. The treatments were distributed in a randomized block design with four replicates. Data were tested by analysis of variance andmeans were compared by Scott-Knott test at a significance level of 5%. The pH values ranged from 3.93 and 4.10. Genotypes differed in all studied nutritional characteristics (p < 0.05). For the acid detergent fiber, differences were detected between the genotypes, AG2501 (35.66%), Volumax (34.89%), AG2005E (34.53%), Qualimax (32.39%) and BRS610 (33.60%). The high participation of lignin was verifiedin all genotypes except for Volumax (4.14%). All silages were within the recommended range 50-65% for in vitro dry matter digestibility and classified as having good quality, however only the AG2005E genotype met nitrogen requirements for microbial fermentation, with 7.06% crude protein. 


1982 ◽  
Vol 242 (6) ◽  
pp. F681-F689 ◽  
Author(s):  
G. D. Stokes ◽  
W. A. Dunson

A study of the permeability of shed epidermis from some terrestrial and freshwater snakes was conducted. Permeability to Br, Na, and K ions was very low and showed a higher influx than efflux in most cases. Permeability to the smaller water molecule was much greater, and in contrast efflux was higher than influx. Skins from aquatic snakes with larger water permeabilities also showed greater permeabilities to Na and K. The highly aquatic Regina septemvittata has the most permeable skin of any snake. Ethanol fluxes were higher than expected for a tracer of its size, perhaps due to its solubility in lipid. Na fluxes through whole live skins of Nerodia cyclopion floridana in vitro were not significantly different from those of the shed skin alone. Isolated hinge regions showed complete water impermeability, suggesting that channels through the skin are located only in the scale region. Dermal water efflux into dry air was considerably less than water-to-water efflux. Lipid extraction increased permeability markedly and eliminated the asymmetry of water and ion fluxes. Lipid replacement with linoleic acid restored half of the water impermeability lost during extraction. Protein extraction did not significantly increase membrane permeability but did eliminate the permeability difference between dry and hydrated skins. Two sizes of lipid-lined channels extending through a protein matrix are suggested as a possible model for snake skin. The diameter of the channels apparently varies in relation to the differing water, Na, and Cl concentrations on opposite sides of the skin, but the mechanism of this adjustment is unknown.


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