PSI-6 Characterization of in vitro stability for two multi-functional processive endoglucanases as exogenous biocatalysts in pig nutrition

Poor efficiency of dietary fibre utilization limits global pork production profit margins and mitigation of footprint on environment. The objective of this study was to characterize in vitro stability for two unique processive endoglucanases of tCel5A1 and p4818Cel5_2A that are reported to hydrolyze natural cellulose and have multi-functionality towards hemicelluloses (Basit and Akhtar, Biotechnology and Bioengineering, 2018, 115:1675; Wang et al., Scientific Reports 2019, 9:13630). Both tCel5A1 and p4818Cel5_2A were modelled with the SWISS-MODEL online server and analyzed and visualized by PyMOL2.4.1. These two cellulases were overexpressed in ClearColi®BL21(DE3). Their endoglucanase activities were determined using 0.70 % sodium carboxymethyl cellulose and 5 mM dithiothreitol (DTT) with or without N2 gas purging; and incubated at pH 6.5–7.0 and 37 °C for 15 min. The three-dimensional models showed 1 and 4 Cys residues on the surfaces of tCel5A1 and p4818Cel5_2A, respectively, suggesting their susceptibility to auto-oxidation by air-borne O2. This prediction was tested by comparing the activities of N2-purged and non-purged enzyme preparations as well as their processing and incubation handling. Both tCel5A1 and p4818Cel5_2A activities were enhanced (P < 0.01) by 50% when 5-mM DTT and N2-purging were adopted. Furthermore, after incubating both tCel5A1 and p4818Cel5_2A enzyme preparations under the porcine gastric pH (3.5) and pepsin (274 U/mL) as well as intestinal trypsin (78 U/mL) and chymotrypsin (20 U/mL) activities at pH 6.5 during 0–5 h, Eadie-Hofstee inhibition kinetic analyses showed that tCel5A1 and p4818Cel5_2A respectively lost 18 and 68% (P < 0.01) of their initial activities after 2 h under the gastric conditions and more than 90% (P < 0.01) of their initial activities after 2–3 h under the intestinal conditions. Therefore, further enzyme protein engineering and/or post-fermentation treatments, such as coating for by-passing the gastric-intestinal environment, will be required to enable these two processive endoglucanases as efficacious exogenous fibre enzymes.

Gas Purging System to Extend Thrust Bearing Life in ESPs

Electric submersible pump (ESP) systems use thrust bearings in the seal section to handle the thrust generated by the pump stages. Thrust bearings are subjected to harsh operating conditions, including high loads, poor oil circulation, and motor oil viscosity degradation. A less-recognized issue is gas becoming centrifugally trapped under the thrust runner. The gas may be present because of incomplete purging of air during filling, permeation of well gas into the motor oil, or gradual gasification of motor oil at high temperatures. Because thrust bearings are such critical components, it is of interest to increase their reliability, which in turn will increase ESP life. A novel gas purging system (GPS) was designed to alleviate stressors on thrust bearings, including gas accumulation, viscosity deterioration and gasification at high temperature, and low working oil volume. GPS circulates oil along with any gas that accumulates under the thrust runner up to a quiet separation chamber. Degassed oil circulates back to the thrust bearing, while accumulated gas eventually purges to the wellbore through relief valves on subsequent on/off cycles. GPS also improves viscosity and reduces gasification by cooling the oil, and it provides a greater working volume of thrust bearing oil to reduce the effects of oil deterioration. This paper details the GPS design principles as well as the optimization of the different design parameters that affect its performance conducted via computational fluid dynamics (CFD). Observations captured on a test fixture built using the final configuration are also presented, validating the intended functionality.

Effect of Nitrogen Content on the Formation of Inclusions in Fe-5Mn-3Al Steels

The effect of N content on the characteristics and formation of inclusions in the Fe-5Mn-3Al steels was investigated in this study. Two synthetic steel melts were produced by two different methods—N2 gas purging and injecting—to introduce nitrogen into the melt. The N content of steel melt varied from 2 to 54 ppm. An increase in the N content to 47 ppm (for 533N-P) and 58 ppm (for 533N-I) increased the total amount of inclusions from 13 to 64 mm−2 and from 21 to 101 mm−2, respectively. The observed inclusions were Al2O3(pure), Al2O3-MnS, AlN(pure), AlN-MnS, AlON, AlON-MnS, and MnS. When the N content was less than 10 ppm, AlN-MnS inclusions were the primary type of inclusions and they formed as solidification products. With an increase in the N content, AlN(pure) inclusions became the dominant type of inclusions as AlN was stable in the liquid steel. These findings were confirmed by thermodynamic calculations. The influence of cooling rate on the types of inclusions was studied and a higher number of AlN-MnS inclusions were observed in samples with slow cooling rate.

Benefits from improved bath agitation with the RADEX DPP gas purging system during EAF highalloyed and stainless steel production

Modern production processes of high alloyed and stainless steel grades are subject to the cost-optimized production of raw steel or molten combined with high flexibility. Excellent mixing of the steel melt helps to improve mass and heat transfer in the EAF, in order to accelerate the melting of heavy scrap and alloys, decarburization, homogeneous superheating, alloy distribution, and to avoid skull formation and solid remainders that may obstruct tapping. The RADEX DPP system generates customer benefits by improving EAF processes during high-alloyed stainless steel making. The most important effect of the enhanced steel bath mixing besides improved heat transfer and decarburization is a decreased chromium oxidation. Firstly, chromium oxidation is decreased by improving the homogeneous distribution of carbon and chromium in the melt. Secondary, build-up of chromium-containing solids in the EAF hearth and skull formation is decreased by stronger melt movement. The cost savings of both effects economically justify the application of gas purging in EAF stainless steelmaking. Case studies are presented demonstrating the advantages. Ill.7. Ref. 7. Tab. 3.

Semi-Solid Slurry Casting Using Gas Induced Semi-Solid Technique to Enhance the Microstructural Characteristics of Al-4.3Cu Alloy

Semi-solid processing of Al-4.3%Cu (A206) alloy was performed by Gas Induced Semi-Solid (GISS) process in different condition. The flow rate of argon gas, starting temperature for gas purging (the temperature of superheated-melt) and the duration of gas purging were three key process variables which were changed during this investigation. It was found that inert gas purging near liquidus, significantly, led to the microstructural modification from fully dendritic to globular structure. Thermal analysis was successfully implemented through CA-CCTA technique to understand the cause of the microstructure change during GISS process. The results showed that gas purging into the melt leads to temperature drop of the melt to its liquidus just after a few seconds from start of gas purging. In fact, copious nucleation was induced by cooling effect of inert gas bubbles. Microstructural features were characterized in semi-solid as well as on conventionally cast samples. The optimum gas purging temperature, injection time, and inert gas flow rate was determined in semi-solid processing to obtain the best globularity in the microstructure of a long freezing range alloy. However, the microstructure of the conventionally cast sample was fully dendritic with shrinkage which affects the soundness of casting products.