scholarly journals Proteomic Examination for Gluconeogenesis Pathway-Shift during Polyhydroxyalkanoate Formation in Cupriavidus necator Grown on Glycerol

2020 ◽  
Vol 7 (4) ◽  
pp. 154
Author(s):  
Nuttapol Tanadchangsaeng ◽  
Sittiruk Roytrakul
Keyword(s):  
Mass Spectrometry ◽  
Glycosyl Hydrolase ◽  
Expression Patterns ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cupriavidus Necator ◽  
Production Costs ◽  
Raw Material ◽  
Short Chain ◽  
Bacterial Cells ◽  
Glucose Content

Because of availability and inexpensiveness, glycerol can be considered as a suitable raw material for polyhydroxyalkanoate (PHA) production with bacterial fermentation. Nevertheless, compared to the production of glucose as a raw precursor, PHA produced from glycerol by Cupriavidus necator was found to produce lower PHA with low bacterial growth rates. According to our study, C. necator was able to synthesize glucose-like intermediates from glycerol via gluconeogenesis. This resulted in a decrease of the cell dry weight and the yield of PHA polymers, especially in the active cell growth phase. It was indicated that glycerol used as a carbon source of the PHA synthesis pathway has glucogenesis-shift, which causes a decrease of the PHA content and productivity. In this research, we investigated the proteins that were closely expressed with the increase of the intracellular PHA and glucose content. For solving the above problem, the proteins inside the bacterial cells were analyzed and compared to the database proteins via mass spectrometry. The proteins were isolated by 1-D SDS-polyacrylamide gel electrophoresis (PAGE) technique and identified by the liquid chromatography mass spectrometry (LC-MS) technique. By using bioinformatics validation, a total number of 1361 proteins were examined and found in the culture bacterial cells. Selective protein expression was correlated with the amount of PHA at each cultivation time and generating glucose by studying the 1361 proteins was elucidated in proteomic information. The results of the cluster of proteins were found to contain 93 proteins using the multiple array viewer (MEV) program with the KMS data analysis model. Protein species with the same expression pattern for PHA and six proteins with similar expression patterns were found to be correlated with generating glucose content. The associations of the two protein groups were then determined through a Stitch program. The protein and chemical associations were analyzed both directly and indirectly through different databases. The proteins of interest were found with research data linked between glycerol and glucose. Five protein types are connecting to glucose and glycerol shift pathway, two of which are glycosyl hydrolase (H16_B1563) and short-chain dehydrogenase (H16_B0687), both of which are enzymes used to break the bonds of complex sugars, possibly related to the partial conversion of glycerol to glucose. The two proteins found in the strains used in the Cupriavidus necator H16 experiment give rise to the break down the bonds of α,α-1,1-glucoside of malto-oligosyltrehalose and short-chain sugar molecules such as mannitol (C6H14O6), respectively. In this research, finding the associated expression proteins which is involved in changing the pathway of gluconeogenesis shift to PHA synthesis will be useful information on genetically modifying microorganisms to produce PHA more efficiently, leading to reduction of the production costs.

Altered protein expression at early-stage rat hepatic neoplasia

2007 ◽  
Vol 292 (5) ◽  
pp. G1272-G1282 ◽  
Author(s):  
Qilie Luo ◽  
Linda Siconolfi-Baez ◽  
Pallavi Annamaneni ◽  
Mark T. Bielawski ◽  
Phyllis M. Novikoff ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Expression ◽  
Immunofluorescence Microscopy ◽  
Early Stage ◽  
Expression Patterns ◽  
Polyacrylamide Gel Electrophoresis ◽  
Normal Liver ◽  
Angiogenesis Inhibitors ◽  
Peptide Mass ◽  
Hepatic Neoplasia

Protein expression patterns were analyzed in a rat model of hepatic neoplasia to detect changes reflecting biological mechanism or potential therapeutic targets. The rat resistant hepatocyte model of carcinogenesis was studied, with a focus on the earliest preneoplastic lesion visible in the liver, the preneoplastic hyperplastic nodule. Expression differences were shown by two-dimensional polyacrylamide gel electrophoresis and image analysis. Polypeptide masses were measured by peptide mass fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) and their sequences were obtained by tandem mass spectrometry. Alterations in expression of cytoskeletal and functional proteins were demonstrated, consistent with biological changes known to occur in the preneoplastic cells. Of particular interest was the differential expression of a serine protease inhibitor (serpin) with a role implicated in angiogenesis. Serpin, implicated in the inhibition of angiogenesis, is present in normal liver but has greatly reduced expression at the preneoplastic stage of liver cancer development. Immunofluorescence microscopy with antibodies to this serpin, kallistatin, supports the proteomic identification. Immunofluorescence microscopy with antibodies to the blood vessel marker von Willebrand factor provides evidence for neovascularization in the liver containing multiple preneoplastic nodules. These observations suggest that at an early stage of liver carcinogenesis reduction or loss of angiogenesis inhibitors may contribute to initiation of neoangiogenesis. A number of other identified proteins known to be associated with hepatomas are also present at early-stage neoplasia.

Graubündens Wald im Fokus | Focus on the Grisons forest

2009 ◽  
Vol 160 (7) ◽  
pp. 195-200
Author(s):  
Reto Hefti
Keyword(s):  
Climate Change ◽  
Global Warming ◽  
Natural Hazards ◽  
Positive Attitude ◽  
Production Costs ◽  
Raw Material ◽  
Intensive Use ◽  
Form Part ◽  
New Challenges ◽  
The Government

In the mountainous canton Grisons, much visited by tourists, the forest has always had an important role to play. New challenges are now presenting themselves. The article goes more closely into two themes on the Grisons forestry agenda dominating in the next few years: the increased use of timber and climate change. With the increased demand for logs and the new sawmill in Domat/Ems new opportunities are offered to the canton for more intensive use of the raw material, wood. This depends on a reduction in production costs and a positive attitude of the population towards the greater use of wood. A series of measures from the Grisons Forestry Department should be of help here. The risk of damage to infrastructure is particularly high in a mountainous canton. The cantonal government of the Grisons has commissioned the Forestry Department to define the situation concerning the possible consequences of global warming on natural hazards and to propose measures which may be taken. The setting up of extensive measurement and information systems, the elaboration of intervention maps, the estimation of the danger potential in exposed areas outside the building zone and the maintenance of existing protective constructions through the creation of a protective constructions register, all form part of the government programme for 2009 to 2012. In the Grisons, forest owners and visitors will have to become accustomed to the fact that their forests must again produce more wood and that, on account of global warming, protective forests will become even more important than they already are today.

SELISIH BIAYA PRODUKSI SEBAGAI ALAT PENGENDALIAN MANAJEMEN PT INDO PUSAKA BERAU

2019 ◽  
Vol 3 (02) ◽  
pp. 397
Author(s):  
SAFITRI NURHIDAYATI ◽  
RIZKI AMELYA SYAM
Keyword(s):  
Raw Materials ◽  
Field Research ◽  
Analytical Tool ◽  
Production Costs ◽  
Raw Material ◽  
Labor Costs ◽  
Material Costs ◽  
Library Research ◽  
The Difference ◽  
The Cost

This study aims to analyze whether the difference that occurs in the cost of raw materials, direct labor, and factory overhead costs between the standard costs and the actual costs in PLTU LATI is a difference that is favorable or unfavorable. Data collection techniques with field research and library research. The analytical tool used is the analysis of the difference in raw material costs, the difference in direct labor costs and the difference in factory overhead costs. The hypothesis in this study is that the difference allegedly occurs in the cost of raw materials, direct labor costs, and factory overhead costs at PT Indo Pusaka Berau Tanjung Redeb is a favorable difference. The results showed that the difference in the cost of producing MWh electricity at PT Indo Pusaka Berau Tanjung Redeb in 2018, namely the difference in the price of raw material costs Rp. 548,029.80, - is favorable, the difference in quantity of raw materials is Rp. 957,216,602, - is (favorable) , the difference in direct labor costs Rp 2,602,642,084, - is (unfavorable), and the difference in factory overhead costs Rp 8,807,051,422, - is (favorable) This shows that the difference in the overall production cost budget is favorable or profitable. This beneficial difference shows that the company is really able to reduce production costs optimally in 2018.  

Technical accounting: sugar extraction and losses

2020 ◽  
pp. 712-721
Author(s):  
Jan Maarten de Bruijn de Bruijn
Keyword(s):  
Process Improvement ◽  
Best Practice ◽  
Continuous Process ◽  
Extraction Yield ◽  
Production Costs ◽  
Raw Material ◽  
Sugar Production ◽  
Cost Leadership ◽  
Sugar Extraction ◽  
White Sugar

The bought sugar in the processed raw material (either beet or cane) comprises a high financial value and may contribute to somewhere around 50% of the white sugar production costs. It is therefore of the utmost importance to minimize sugar losses along the process and produce as much white sugar as possible from the raw material. This paper explains the principle of technical accounting as tool to control sugar extraction and losses in beet sugar manufacture. The sugar mass balance used to calculate the overall sugar extraction yield, as well as several simple calculations proposed for estimating the different sugar losses (like e.g. extraction (diffusion) losses, infection losses, sugar losses in molasses, etc.) in the subsequent process steps will be explained in detail. Proper technical accounting is considered indispensable for continuous process control and process improvement in pursuit of best-practice operation and cost-leadership.

Mass Spectrometry-Based Identification of Urinary Antimicrobial Peptides in Dairy Cows

2020 ◽  
Vol 27 (3) ◽  
pp. 225-235
Author(s):  
Ambika Sharma ◽  
Rajesh Nigam ◽  
Ashish Kumar ◽  
Simmi Singh
Keyword(s):  
Mass Spectrometry ◽  
Antimicrobial Peptides ◽  
Polyacrylamide Gel Electrophoresis ◽  
Radial Diffusion ◽  
Protein Database ◽  
Antimicrobial Effects ◽  
Cow Urine ◽  
Radial Diffusion Assay ◽  
Promising Source ◽  
Diffusion Assay

Background:: Urine is considered one of the biological fluids in which antimicrobial peptides are secreted or expressed. Cow urine has not been investigated for the presence of these peptides using MALDI-TOF-MS. Objective:: The aim of this study is to isolate, identify and assess the antimicrobial activity of urinary antimicrobial peptides from healthy normal cycling cows. Method:: We analyzed the urine sample using diafiltration, ion exchange chromatography, Reverse Phase High-Performance Liquid Chromatography (RP-HPLC), acid urea polyacrylamide gel electrophoresis (AU-PAGE) coupled with identification through Peptide Mass Fingerprinting (PMF) by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDITOF- MS). The in vitro antimicrobial effects of purified fractions were assessed using Radial Diffusion Assay (RDA) and microtitre broth dilution assay against Gram-positive and Gramnegative bacteria. Results: : Proteins corresponding to the peaks were identified using SWISSPROT protein database. This study revealed constitutive expression of β-Defensin-1 (DEFB1), β-Defensin-4A (DFB4A), Neutrophil Defensin-1 (DEF1), Neutrophil Defensin-3 (DEF3) in cow urine. The identified peptides are cationic antimicrobial peptides of the defensin family. The purified fractions exhibited antimicrobial effects in radial diffusion assay and MIC values in the range of 2.93-29.3 &*#181;M/L. Conclusion:: This study concludes that cow urine, previously unexplored with regard to antimicrobial peptides, would be a promising source of highly potent AMPs and an effective alternative to the resistant antibiotics.

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