The Weight Molecular Weight Of Pectin

In this experiment, 4 grams of peeled turnip was used to prepare the enzyme extract opposed to the 1 gram of turnip suggested by Fundamentals of Life Science. Along with the change to the amount of turnip used, the amount of 0.1M phosphate buffer used to prepare the enzyme extract was changed from 50mL to 30mL. The affect of temperature on enzyme activity was not

The results for Benedict’s test for reducing sugars before hydrolysis, the control dH2O had no color change, as well as sucrose and raffinose. Gelatin became a dark blue and egg albumin a light greyish blue. Glucose yielded a dark orange, milk albumin turned orange, and starch had a yellow precipitate. In the Benedict’s test for reducing sugars by hot acid hydrolysis, the control dH2O was blue or no reaction. Glucose turned a brownish-orange; sucrose, a reddish-brown, raffinose, light pink; starch, a dark yellowish-orange; gelatin, violet; milk powder, a light yellow; and egg albumin, a greyish-violet. In the Lugol’s test for polysaccharides, the control dH2O turned yellow or no reaction, as well as glucose, sucrose, raffinose, milk powder and albumin. Starch was clear with blue-violet on the bottom (precipitate) and gelatin turned a slightly opaque white. In the test for polysaccharides remaining after hydrolysis with Lugol’s solution the control dH2O was yellow or no reaction. Glucose, sucrose, raffinose, starch and gelatin also had no reaction. Milk powder had a white precipitate and particle suspension. Egg albumin had an even greater white precipitate and particle suspension. In Biuret’s test for proteins there was no reaction for dH2O, glucose, sucrose, raffinose, and starch. Gelatin, milk powder and egg albumin all turned slightly violet with bubbles.

In Conclusion pectinase would be the better enzyme to use, I found that the more pectinase used in the applesauce the more apple juice was produced. The hypothesis is supported by the data. The trial for pectinase produced 13ml of apple juice. The trial for cellulase produced 8ml. The trial for water produced 5ml .A reason why this lab could have not worked is because there may have been too much enzymes dropped or too much water dropped in the applesauce. A suggestion I would give to make this experiment more efficient would to be able to filter the applesauce and pay more attention to the time and be careful when the enzymes are dropped into the apple juice also by making the time longer and by doing more than 1 trial. Testing the enzymes

The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the

Protein Assay: The Pierce BCA Protein Assay (Thermo Scientific) is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein concentration. A series of standard solution of Bovine Serum Albumin (BSA) ranging from 0-2000 µg/ml was prepared from a stock solution of 2 mg/ml BSA. 25ul of diluted crude (1:500, 1:250), desalted (1:100, 1:50), and 6 peak fractions from cibarcon blue column (1:10, 1:5) were loaded in microplate along with 175ul of BCA working reagent. Microplate was incubated for 30min at 370C and then the absorbance was measured at 562nm.

vulgaris plants, via the formation of a standard curve prepared using varying concentrations of bovine serum albumin (BSA) solution. Following absorbance readings of the various BSA solutions, they were plotted against their concentrations providing an indirect measure for determining protein concentrations of the plant samples within the assay tubes, and through further calculations the sample protein concentration. The mean protein concentration for the control group was calculated to be 3.34 ± 1.30 mg/mL, while the mean treated group concentration was 2.01 ± 1.26 mg/mL. These results similarly like the chlorophyll results correlate with the literature articles, as a reduced protein content within the Paraquat treated plants can be expected to some extent (Chia et al., 1981). This reduction in protein concentration is the result of those superoxide anions produced by Paraquat, disrupting the chloroplast membranes and allowing for intracellular components including some proteins to leak out, hence the decrease in protein concentration in comparison to the non-treated plants (Qian et al., 2009). A slight outlier may exist within the treated groups protein concentrations as one of the groups provided a negative value for protein concentration which is not valid, but even after exclusion of that data value, results are still supportive of the expected outcome. Though these results support the claim of Paraquat toxicity causing membrane deterioration and leakiness, protein concentration values are rather more purposeful when used to analyze malondialdehyde (MDA) values on per mg of protein

Rubisco is the most abundant protein on earth that is essential for carbon fixation in plants. For the protein to function at its optimal level, it needs to be isolated from the mixture of proteins and in its purest form. The three isolation techniques carried out in this lab are salting out, ion exchange chromatography, and SDS-PAGE. Rubisco will be purer as each technique is conducted and will be in its purest form after the last isolation technique is carried out.

1H Nuclear magnetic resonance (NMR) of hydrolysed products obtained by enzyme CtXynGH30 treatment of substrate beechwood xylan was performed using a spectrometer (Bruker, 600 MHz). The enzyme reaction was performed in 200 μl reaction mixture by taking 3.4 μg of CtXynGH30 and (1%, w/v) beechwood xylan dissolved in 20 mM sodium phosphate buffer, pH 6.0 and incubated for 1 h at 50°C. The protein and undigested polysaccharides were precipitated by adding equal volume of acetone (200 μl) with gentle mixing and subsequently centrifuged at 13000g for 5 min. The supernatant was lyophilized. The 10-15 mg of this lyophilized sample was dissolved in the 0.75 ml of D2O. The sample was placed in the NMR tube (0.5 mm, Sigma-Aldrich) and the spectra were

From the stock substrate solution of 2.5 mM, each group serially diluted at least one different substrate concentration for a total of four different substrate concentrations to be investigated: 1.25 mM, 1.0 mM, 0.75 mM, 0.25 mM. The enzyme concentration was kept constant at 2.0 mM while experimenting on the affect of varying enzyme concentration on the rate and product formation of ONP. Enough 2.0 mM enzyme solution was prepared in the previous part of the project to supply this assay. Using similar procedure to collect absorbance data as the first part, 0.5 mL of 2.0 mM enzyme concentration was placed into the cuvette and used to calibrate the spectrometer at 420 nm. Data was then started, with the immediate addition of 0.5 mL of varying substrate concentrations. Each varying substrate concentration was split between the team and run for a total of 10 minutes, with the exception of the 1.25 mM run. Upon completion, data from each varying substrate concentration was copied to a single Excel sheet and used to produce an absorbance vs. time graph, product formation vs. time graph, Michaelis Menten plot, and Lineweaver-Birk plot. This analysis was used to calculate the V0,Vmax, and Km for β-Galactosidase

, on the other hand, polymer concentration had negative correlation with sonolysis. Sweet potato pectin posseed good antioxidant activity which was enhanced by sonication process. Moreover, this research showed the potential application of ultrasonic in pectin depolymerization, which could also be applied in other polymers. Further study should be carried out to elucidate the degraded pectin materials composition and structure, its potential applications in plant protection, antimicrobial, anticancer, prebiotic

3.8.6. The influence of some metal ions and some chemical compounds on the purified pectinase activity:

Probiotic or drug viability can be optimized by providing a physical barrier separating unfavourable conditions from the delicate cargo. Furthermore, calcium alginate demonstrates non-toxic properties deeming it worthy as a promising delivery agent for probiotics or drugs. On the other hand, limitations associated with the experiment performed include; lack of a secondary or multi- additional coating, texture, and flavour of the polymer. Additionally, limitations associated with simulated stomach acid involve the absence of peristalsis; a primary component of smooth-muscle movement through-out the digestive system. Future applications in regards to calcium alginate may one day implement the use of the biopolymer for; probiotic and drug delivery, wound-sealing, probiotic dressings, and perhaps tissue engineering. In conclusion, calcium alginate exhibits exceptional protective properties from chemical or mechanical factors and holds promise as an effective encapsulation technology for the delivery of probiotics and perhaps drug

It is mainly used for the detection of fatty acids in the biosurfactant. Fatty acid like hexadecanoic acid, octanoic acid and decanoic acid are identified by GC-MS analysis (Sadouk et al., 2008). Some studies revealed the presence of fatty acids like palmitic acid and oleic acid (Bayoumi, 2010). For GC-MS analysis carbohydrates are subjected to trimethylsilation (TMS) (Dejongh et al., 1969). Sugars like trehalose, sophorose and glucopyranose are also identified during these analyses. Colorimetric method was carried out to preliminary analyze chemical compositions of crude lipopeptides. Total lipid, proteinand sugar contents were determined by sulfo–phospho–vanillintest (Izard et al,. 2003), Bradford assay (Satpute et al,. 2010) and phenol–sulfuric acid test (DuBois et al,. 1956) respectively. The fatty acid contents of the lipid in crude lipopep-tides were analyzed by gas chromatography (GC) after fatty acidmethyl ester (FAME) derivatization (Silva et al,.

increasing power led to decreasing molecular weight (Fig. 2a). Polydispersity did not show consistent trend with the increasing power, which corresponded to random scission of pectin (Fig. 2b). Pectin molecular weight decreased rapidly between 100W to 200W, however,the molecular weight reduction reduced between 200W to 400W, which is consistent with increased number of molecules and presence of small chain and stiff molecules in the system which tend to be less prone to sonolysis. The 400W power treatment resulted in more than four fold decrease in molecular weight compared to native pectin, a similar observation was made by Zhang et al., [46] who reported progressive decrease in citrus pectin molecular weight with increasing ultrasonic intensity while. Sonolysis of chitosan under different sonic intensities lead to decrease in molecular weight with increasing ultrasonic power, suggesting that decrease in sonolysis rate with increasing power was likely caused by increase in total number of molecules and presence of low molecular weight pectin fragments which are stiff, hence much higher energy is required to further reduce their molecular size [24]. Increased sonication intensity led to enhanced molecular weight reduction in polylactic acid (PLA) and Poly-lactic/glycolic acid (PLGA) [47]. Moreover, sonolysis of chitosan produced low molecular weight product with increased bioactivity.

Different techniques and principles for protein extraction and characterization were demonstrated in this experiment. Various proteins were extracted from different sources: 1.67 g yeast invertase, 1.03 g egg white albumin, and 5.15 g of milk casein. Activity assay for invertase was performed using Benedict’s test and the enzymes inverting action on sucrose was confirmed. Warburg-Christian Method and Bradford Assay were also employed to determine the protein concentration in the albumin and the casein samples. The concentrations for the albumin and casein samples were found to be 0.519 and 0.327 mg/mL, respectively based on Warburg-Christian Assay; and 6.5x10-3¬ and 1.9x10-2 mg/mL