Globulin expression in grain of Australian hard wheat cultivars is affected by growth environment

Our aim was to study changes in wheat proteomes across different growth locations as the first step in linking protein composition with functional changes in grains produced using commercial production systems. Soluble and insoluble proteins were extracted sequentially from grain of three commercial wheat cultivars grown at four locations in New South Wales, Australia during a single season. Bands were separated using SDS-PAGE and identified by peptide mass fingerprinting. Quantitative changes in the electrophoretic patterns were observed mainly in the insoluble polypeptides of molecular mass between 40 – 70 kDa for all three cultivars grown at two of the four locations. These proteins were identified as mainly globulin and serpin isoforms, and triticin. Other proteins with changed expression included disease-resistance proteins, class III peroxidase, starch branching enzyme I, β-amylase, and storage proteins. Two-dimensional electrophoretic analysis was performed on two of the same wheat cultivars grown at one of the locations during two consecutive seasons. Protein spots that varied between seasons consisted of globulin and serpin isoforms, triticin, HMW-glutenin, gamma-gliadin, starch branching enzyme IIb and alpha amylase. The implications of the upregulation of globulin and triticin on whole meal flour quality, through their participation in polymerization of the gluten network, are considered

ALPHA-AMYLASE INHIBITORY ACTIVITY OF INHIBITOR PROTEINS IN DIFFERENT TYPES OF COMMERCIAL RICE

The inhibition of α-amylase enzyme by the inhibitor proteins extracts of all the three types of commercial rice, white rice (WR), brown rice (BR) and glutinous rice (GR) were investigated. Among the three types of rice, the BR has significantly the highest concentration of the inhibitor protein (0.030 ± 0.002 mg/ml) compared to glutinous rice (0.006 ± 0.001 mg/ml) and white rice (0.005 ± 0.001 mg/ml). In term of the percentage of inhibition of the α-amylase enzyme of the Aspergillus oryzae, the brown rice shows the highest inhibition (61.22 %) among the three types of rice with the lowest maltose liberated. The acarbose (0.1 mg/ml) which was used as a positive control represent the highest inhibition of the α-amylase enzyme inhibitor among all of the tested samples. Even though inhibitor protein extract of BR is possessed lower ability to inhibit the α-amylase, yet it still can be one of the best option and alternative for the Diabetes Mellitus patients for their daily consumption compared to the WR and GR

ANTI-DIABETIC POTENTIAL OF PEPTIDE FROM P. NIRURI REVEALS THROUGH CARBOHYDRATE HYDROLYZING ENZYME INHIBITION ASSAY

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anti-diabetic properties. In this research, protein extracts from different plant parts (leave, fruit and stem) of P. niruri were investigated for their anti-diabetic potential through α-amylase and α-glucosidase enzyme inhibition assays. For the enzyme inhibition assay, fruit was found to have the highest inhibition percentage (90.0%) against α-glucosidase followed by leave (62.6%) and stem (38.4%). The similar patterns were also recorded for the α-amylase enzyme inhibition assay, in which, fruit showed the highest inhibition percentage (64.1%) followed by leave (33.3%) and stem (18.2%). The findings of this research suggest that fruit of P. niruri is a potential plant part with regards to anti-diabetic properties since it exhibits the highest inhibition activity against both α-amylase and α-glucosidase enzyme compared to leave and stem

Effect of high inlet temperature of spray dryer on viability of microencapsulated Trichoderma asperellum conidia

In recent years, the use of spray drying for microencapsulating beneficial microbes has gained the interest of researchers, mainly due to dried powder formulation could prevent contamination and prolong self-life of the microbes. The major constraint of spray drying is conidia could lose viability during the drying process due to heat. In this present study, the effect of spray drying inlet temperature on viability of microencapsulated Trichoderma asperellum conidia was assessed. A blend biopolymer of gum Arabic and maltodextrin in ratio of 1:1 was used to microencapsulate the conidia at 150, 160 and 170oC inlet temperatures of spray drying process. Assessment of conidia viability was performed based on conidia percent survival of spray dried (PSsd) and survival increase (SI) unit values. Viability of the microencapsulated conidia was also evaluated their shelf life stored at two different temperatures which were at 28 ±2˚C and 4±2˚C for 40 weeks. The finding showed that viability of microencapsulated conidia was optimum obtained at inlet temperature of 170˚C with 68.2% of PSsd and SI and 17.7 units of SI compared to 15.9% and 1.5 unit respectively obtained for 160oC and 0.2% and 0.7 unit for 150oC. The highest inlet temperature has showed the highest viability compared to lower temperatures. Conidia stored at low temperature of 4 ±2˚C has survived longer up to 40 weeks that were confirmed via the viability test. High inlet temperature of 170oC was desirable to enhance survivability and viability of the conidia to be used as biocontrol agent up to 40 weeks at low temperature storage. These microencapsulated conidia could be further tested on their capability to inhibit the pathogen of pineapple diseas

Functional annotation of selected streptococcus pneumoniae hypothetical proteins

The ability of Streptococcus pneumoniae to induce infection relies on its virulence factor machinery. A previous study has identified essential proteins that might be responsible towards the pathogenicity of S. pneumoniae serotype 2 strain D39. How ever, 39 of them were yet functionally and stru cturally uncharacterized. Thus, by using in silico approach, this study aims to annotate the function and the structure of these unannotated proteins. Initially, all 39 targeted proteins went through primary screening for template availability and pathogen icity. From there, 11 of them were selected and were further analyzed on the basis of their physicochemical, functional and structural categorization using an integrated bioinformatics approach by means of amino acid sequence and stru cture - based analysi s. The obtained data suggested that all targeted proteins showed high possibility to be involved in either cell viability or cell pathogenicity mechanism of the bacterium, with SPD_1333 and SPD_1743 being the two most promising proteins to be further studied. Findings from this study could provide a better understanding of the pathogenic ability of this microorganism and thus, enhance drug development and target identification processes in the aim of improving pneumococcal disease control

Proteomic analysis of rice seed storage proteins in relation to nutrient quality of three different commercial rice types

Seed storage proteins (SSPs) are the most important component in rice which provides nutrient to consumers. The SSPs content and composition are among the important determinant for rice quality determination specifically for nutritional value. Multiple factors have been identified to give effect to the nutritional value of rice grain including different types of rice. The purpose of this study is to investigate the expression level of SSPs of rice and also relates the nutrient quality of rice with the variability in SSPs expression from three different types of rice commercially available in markets. The SSPs were extracted from three different types of rice; white rice, brown rice and glutinous rice of the local rice brands in markets. Bradford Assay was carried out to determine the total SSPs content and brown rice was found to have significantly higher total SSPs content (9.157± 0.4 mg/100mg seeds) compared to white rice (6.933± 0.8 mg/ 100mg seeds) and glutinous rice (5.388± 0.2 mg/ 100mg seeds). Based on the banding patterns of the SDS-PAGE, there were different level of expressions were observed between the three rice types. The different expression can be observed obviously at the glutelin precursor region and glutelin acidic subunit region and prolamin polypeptides region for each type of rice. Therefore, contribute to the different nutritional value for dietary intake

Development of a streamlined recombinant protein production and purification protocols from selected bacterial species applicable for biochemical, biophysical and structural analyses

Researches using recombinant protein from pathogenic bacterial samples are widely used to understand the biochemistry and processes used by the specific microbes for its viability or ability to evade human host immune system. The specific protein must first be isolated from the bacteria. Cloning the gene encoding the protein, followed by expression and purification in a suitable bacterial host will allow us to obtain a protein purified to suitable level for subsequent biochemical and biophysical experiments. IIUM Kuantan Campus, to the best of our knowledge, does not have a laboratory completely set up for recombinant protein production and purification from bacterial samples. This project aims to develop a suitable laboratory facility and protocol that can be utilized routinely for this purpose. We will clone genes from selected bacterial species with specific known annotated function in a recombinant plasmid; and a fusion tag will be added to help in isolating the protein of interest. Escherichia coli will be used as the bacterial host for expression and purification trials. This project is expected to provide proof of concept for the feasibility of routine cloning, expression and purification of recombinant bacterial protein in our Protein Laboratory suitable for further biochemical, biophysical or structural studies

Anti-diabetic potential of peptide from P. niruri reveals through carbohydrate hydrolyzing enzyme inhibition assay

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anti-diabetic properties. In this research, protein extracts from different plant parts (leave, fruit and stem) of P. niruri were investigated for their anti-diabetic potential through α-amylase and α-glucosidase enzyme inhibition assays. For the enzyme inhibition assay, fruit was found to have the highest inhibition percentage (90.0%) against α-glucosidase followed by leave (62.6%) and stem (38.4%). The similar patterns were also recorded for the α-amylase enzyme inhibition assay, in which, fruit showed the highest inhibition percentage (64.1%) followed by leave (33.3%) and stem (18.2%). The findings of this research suggest that fruit of P. niruri is a potential plant part with regards to antidiabetic properties since it exhibits the highest inhibition activity against both α-amylase and α-glucosidase enzyme compared to leave and stem

Proteomic analysis of bioactive peptide from different plant parts of Phyllanthus Niruri (Dukung Anak) and its anti-diabetic propertes

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties. In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa

Protein profiling of different plant tissues from herb Phyllanthus Niruri

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties. In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa