• Analytical Science and Technology
• /
• v.30 no.2
• /
• pp.96-101
• /
• 2017

This study investigated the effect of centrifugation on tryptic digestion. This was done by applying different centrifugation speeds (6,000, 8,000, 10,000, 20,000, and $30,000{\times}g$) over various durations (0, 10, 20, 30, 40, 50, and 60 min) to digest two model proteins - cytochrome c and myoglobin. The intact proteins and resulting peptides were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Centrifugation greatly improved the tryptic digestion efficiency of cytochrome c, where either an increase in centrifugation speed or in digestion duration significantly improved the digestion of cytochrome c. However, centrifugation did not noticeably improve the digestion of myoglobin; 16 h of centrifuge-assisted tryptic digestion at $30,000{\times}g$ barely removed the myoglobin protein peak. Similar results were also obtained when using conventional tryptic digestion with gentle mixing. When acetonitrile (ACN) was added to make 10% ACN buffer solutions, the myoglobin protein peak disappeared after 6 h of digestion using both centrifuge-assisted and conventional tryptic digestions.

• Mass Spectrometry Letters
• /
• v.4 no.2
• /
• pp.25-29
• /
• 2013

Proteomics for biomarker validation needs high throughput instrumentation to analyze huge set of clinical samples for quantitative and reproducible analysis at a minimum time without manual experimental errors. Sample preparation, a vital step in proteomics plays a major role in identification and quantification of proteins from biological samples. Tryptic digestion a major check point in sample preparation for mass spectrometry based proteomics needs to be more accurate with rapid processing time. The present study focuses on establishing a high throughput automated online system for proteolytic digestion and desalting of proteins from biological samples quantitatively and qualitatively in a reproducible manner. The present study compares online protein digestion and desalting of BSA with conventional off-line (in-solution) method and validated for real time sample for reproducibility. Proteins were identified using SEQUEST data base search engine and the data were quantified using IDEALQ software. The present study shows that the online system capable of handling high throughput samples in 96 well formats carries out protein digestion and peptide desalting efficiently in a reproducible and quantitative manner. Label free quantification showed clear increase of peptide quantities with increase in concentration with much linearity compared to off line method. Hence we would like to suggest that inclusion of this online system in proteomic pipeline will be effective in quantification of proteins in comparative proteomics were the quantification is really very crucial.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.1
• /
• pp.110-119
• /
• 2016

High-solid anaerobic digestion of sewage sludge achieves highly efficient volatile solid reduction, and production of volatile fatty acid (VFA) and methane compared with conventional low-solid anaerobic digestion. In this study, the potential mechanisms of the better performance in high-solid anaerobic digestion of sewage sludge were investigated by using 454 high-throughput pyrosequencing and real-time PCR to analyze the microbial characteristics in sewage sludge fermentation reactors. The results obtained by 454 highthroughput pyrosequencing revealed that the phyla Chloroflexi, Bacteroidetes, and Firmicutes were the dominant functional microorganisms in high-solid and low-solid anaerobic systems. Meanwhile, the real-time PCR assays showed that high-solid anaerobic digestion significantly increased the number of total bacteria, which enhanced the hydrolysis and acidification of sewage sludge. Further study indicated that the number of total archaea (dominated by Methanosarcina) in a high-solid anaerobic fermentation reactor was also higher than that in a low-solid reactor, resulting in higher VFA consumption and methane production. Hence, the increased key bacteria and methanogenic archaea involved in sewage sludge hydrolysis, acidification, and methanogenesis resulted in the better performance of high-solid anaerobic sewage sludge fermentation.

• Journal of Korean Society on Water Environment
• /
• v.26 no.5
• /
• pp.810-815
• /
• 2010

Ultrasonic sludge pre-treatment has been studied to enhance the performance of anaerobic digestion by increasing sludge hydrolysis which is regarded as the rate-limiting-step of anaerobic digestion. In this study, the effect of ultrasonic pre-treatment on sludge hydrolysis (solubilization) and methane production was investigated. Sludge solubilization efficiency increased with ultrasonic energy input. However, it is uneconomical to apply more than 720 kJ/L as the solubilization efficiency per energy input declines afterwards. Volatile fatty acids concentration increased after the ultrasonic sludge hydrolysis. Anaerobic batch digestion showed that methane volume reached 64.7 and 84.5 mL after 18 days of incubation with the control sludge and ultrasonically hydrolyzed sludge, respectively. Methane production potential, maximum methane production rate, and the lag time of modified Gompertz equation were changed from 70 mL, 6.4 mL/day, and 1.2 days to 89 mL, 9.6 mL/day, and 0.5 day, respectively, after the ultrasonic sludge treatment. The results proved that ultrasonic pre-treatment contributed significantly not only for the methane production but also for the reduction of anaerobic digestion time which is critical for the performance of anaerobic sludge digestion.

• Environmental Engineering Research
• /
• v.25 no.1
• /
• pp.1-17
• /
• 2020

With benefits to the human health, environment, economy, and energy, anaerobic digestion (AD) systems have attracted remarkable attention within the scientific community. Anaerobic digestion system is created from (bio)reactors to perform a series of bi-metabolism steps including hydrolysis/acidogenesis, acetogenesis, and methanogenesis. By considering the physical separation of the digestion steps above, AD systems can be classified into single-stage (all digestion steps in one reactor) and multi-stage (digestion steps in various reactors). Operation of the AD systems does not only depend on the type of digestion system but also relies on the interaction among growth factors (temperature, pH, and nutrients), the type of reactor, and operating parameters (retention time, organic loading rate). However, these interactions were often reviewed inadequately for the single-stage digestion systems. Therefore, this paper aims to provide a comprehensive review of both single-stage and multi-stage systems as well as the influence of the growth factors, operating conditions, and the type of reactor on them. From those points, the advantages, disadvantages, and application range of each system are well understood.

• Analytical Science and Technology
• /
• v.17 no.4
• /
• pp.363-367
• /
• 2004

A simple and convenient digestion method was developed for the determination of heavy metal in polyurethane sample using only nitric acid in open system without heating. This digestion method had prevented loss of volatile elements of Hg and As and reduced the pretreatment time for the sample digestion. The detection limit is lowered over 10 times than that of microwave digestion method. Recoveries in polyurethane material were 108% for As, 96 ~ 99% for Hg, 102 ~ 103% for Cr, 98 ~ 102% for Pb, 86 ~ 90% for Cd, and 90 ~ 93% for Cu.

• Journal of Environmental Science International
• /
• v.10 no.1
• /
• pp.13-19
• /
• 2001

An on-line digestion system based on the flow injection analysis technique was presented for the determination of total phosphorus. The system converts condensed and organic phosphoruses into orthophosphate that can be analyzed spectrophotometically. The reaction between orthophosphate and ammonium molybdate in a strong acid medium forms the yellow complex of phosphomolybdate which is next reduced to a molybdic blue complex by stannous chloride. The quantitation of orthophosphate is based on the absorbance of the molybdic blue. To determine total phosphorus, the digestion system was installed between the sample injector and the reaction coil with the added lines of digestion solution(potassium persulfate+sulfuric acid) and the $H_2O$ carrier. The system has shown that the digestion efficiencies were greater than 95% for the typical condensed and organic phosphoruses. When tripolyphosphate standards were used, the calibration data showed that the linear dynamic range extended to a concentration of 1.5ppm with the detection limit of 25ppb total phosphorus. The typical relative standard deviation was less than 2%. The proposed system was successfully applied to lake water, wastewater, and streamwater. The analytical variables such as digestion efficiency, analysis time, and reproducibility were evaluated and compared with the manual digestion method.

• Journal of Animal Science and Technology
• /
• v.65 no.1
• /
• pp.32-56
• /
• 2023

This review explores the factors that improve meat protein digestibility and applies the findings to the development of home meal replacements with improved protein digestion rates in older adults. Various methods improve the digestion rate of proteins, such as heat, ultrasound, high pressure, or pulse electric field. In addition, probiotics aid in protein digestion by improving the function of digestive organs and secreting enzymes. Plant-derived proteases, such as papain, bromelain, ficin, actinidin, or zingibain, can also improve the protein digestion rate; however, the digestion rate is dependent on the plant enzyme used and protein characteristics. Sous vide processing improves the rate and extent of protein digestibility, but the protein digestion rate decreases with increasing temperature and heating time. Ultrasound, high pressure, or pulsed electric field treatments degrade the protein structure and increase the proteolytic enzyme contact area to improve the protein digestion rate.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.2
• /
• pp.599-604
• /
• 2011

In typical proteomic analysis, trypsin digestion is one of the most time-consuming steps. Conventional proteomic sample preparation methods use an overnight trypsin digestion method. In this study, we compared high-pressure cycling technology (PCT) during enzyme digestion for proteome analysis to the conventional method. We examined the effect of PCT on enzyme activity at temperatures of 25, 37, and $50^{\circ}C$. Although a fast digestion (1 h) was used for the standard protein mixture analysis, the PCT-assisted method with urea showed better results for protein sequence coverage and the number of peptides identified compared with the conventional method. There was no significant difference between temperatures for PCT-assisted digestion; however, we selected PCT-assisted digestion with urea at $25^{\circ}C$ as an optimized method for fast enzyme digestion, based on peptide carbamylation at these conditions. The optimized method was used for stem cell proteome analysis. We identified 233, 264 and 137 proteins using the conventional method with urea at $37^{\circ}C$ for 16h, the PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, and the non-PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, respectively. A comparison of these results suggests that PCT enhanced the enzyme digestion by permitting better access to cleavage sites on the proteins.

• Journal of Korean Society of Water and Wastewater
• /
• v.25 no.4
• /
• pp.611-616
• /
• 2011

The impacts of ultrasonic pretreatment on the biodegradability of domestic sewage sludge were evaluated through a series of anaerobic digestion experiments in batch system. The gas and methane production from the sludge samples pretreated by an ultrasonic tool with different durations were measured with time. Although the biogas production increased with the extent of sludge solubilization and the period of ultrasonic pretreatment, the enhancement of sludge biodegradability was much more sensitive to the pretreatment for the relatively short periods. Most of the enhanced biodegradability by the pretreatment was appeared in the early stage of anaerobic digestion, less than 6 days. The maximum biogas production per day was observed between 4 to 6 days when the sludge was pretreated less than 10 minutes while it was obtained in the beginning for the sludge pretreated longer periods. The results suggest that the repeated alternation of low strength ultrasonic pretreatment and anaerobic digestion may be more effective than the combination of one time pretreatment for a relatively long period and following anaerobic digestion.