Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus.

Thermostable alkaline serine protease gene of Geobacillus stearothermophilus B-1172 was cloned and expressed in Escherichia coli BL21 (DE3) using pET-22b(+), as an expression vector. The growth conditions were optimized for maximal production of the protease using variable fermentation parameters, i.e., pH, temperature, and addition of an inducer. Protease, thus produced, was purified by ammonium sulfate precipitation followed by ion exchange chromatography with 13.7-fold purification, with specific activity of 97.5 U mg(-1) , and a recovery of 23.6%. Molecular weight of the purified protease, 39 kDa, was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 90 °C at pH 9. The enzyme activity was steady in the presence of EDTA indicating that the protease was not a metalloprotease. No significant change in the activity of protease after addition of various metal ions further strengthened this fact. However, an addition of 1% Triton X-100 or SDS surfactants constrained the enzyme specific activity to 34 and 19%, respectively. Among organic solvents, an addition of 1-butanol (20%) augmented the enzyme activity by 29% of the original activity. With casein as a substrate, the enzyme activity under optimized conditions was found to be 73.8 U mg(-1) . The effect of protease expression on the host cells growth was also studied and found to negatively affect E. coli cells to certain extent. Catalytic domains of serine proteases from eight important thermostable organisms were analyzed through WebLogo and found to be conserved in all serine protease sequences suggesting that protease of G. stearothermophilus could be beneficially used as a biocontrol agent and in many industries including detergent industry.

Greening for greater good: investigating the critical factors for customer satisfaction with sustainable e-banking.

This study investigated the impact of efficiency, reliability, responsiveness, safety, and security on customer satisfaction with sustainable e-banking. The study used a quantitative methodology, with data collected from customers of commercial banks in South Punjab, Pakistan. A Likert scale questionnaire was used to collect cross-sectional data. A sample of 350 was analyzed using the Statistical Package for the Social Sciences (SPSS) for testing descriptive statistics, correlation analysis, and regression results. The study's results revealed that efficiency, reliability, responsiveness, safety, and security significantly impacted customer satisfaction with e-banking services. Efficiency and reliability were the most crucial factors influencing customer satisfaction, followed by safety, security, and responsiveness. The research findings have important implications for commercial banks, as they highlight the critical role of these factors in the development of sustainable e-banking practices. Overall, the study provides valuable insights for commercial banks seeking to improve their e-banking services and ensure customer satisfaction. Commercial banks can enhance their competitiveness and achieve sustainable growth in the highly competitive e-banking sector by prioritizing efficiency, reliability, responsiveness, and safety and security.

The role of unsustainable HR practices as illegitimate tasks in escalating the sense of workplace ostracism.

Unsustainable HR practices impose illegitimate tasks on employees due to a shortage of resources. These illegitimate tasks bring counterproductive work behavior in terms of workplace incivility that creates a sense of ostracism in employees. To address these issues, the study examined the relationship among unsustainable HR practices in terms of illegitimate tasks and workplace ostracism. Whereas workplace incivility is defined as an underlying reason through which this association exists. Adopting a theoretical framework from earlier research, the study used cross-sectional data and subsequently a method of quantitative research, and the sample comprised permanent faculty members of private universities in Pakistan working in different departments with different role titles. Smart PLS was applied to run multiple statistics analyzed on the obtained data. The results from the study supported the hypothesis by depicting a positive and significant association between illegitimate tasks and workplace ostracism. Further, workplace incivility was playing the mediating role between illegitimate tasks and workplace ostracism. The results from the study carry significant implications for managers and researchers. Recommendations and future research directions are also discussed in the paper.

Exploring the generic skills required for the employability and professional wellbeing of Pakistani Millennials: The employers' perspective.

Introduction: This study aims to elucidate the employers' perspectives on the key generic employability skills which would enable them to seek graduate jobs and will ensure their professional wellbeing once they enter the job market after acquiring a University degree. Methods: In order to pursue this objective, an exploratory qualitative inquiry was deployed which involved two panel-based discussions. The respondents of each panel discussion were invited through the platforms of the Bahawalpur Chamber of Commerce and Industry (BCCI), and the Chamber of Commerce Rahim Yar Khan. These respondents represented Pharmaceuticals, Agrichemical manufacturers (fertilizers, pesticides), Livestock, Cotton, textiles, and apparel industry which are the main industries in Southern Punjab, Pakistan. Results and discussion: For the purpose of analysis, a thematic analysis was done in the context of grounded theory. Resultantly, the findings of the study reveal an employers' perspective on 10 key generic employability skills as must-have for a Pakistani Millennial graduate including various soft and hard skills, such as-Emotional Intelligence, Flexibility and Adaptability, Multitasking, Computer Literacy and Digital Skills, Information Literacy and Data Analytics, Oral and Written Communication in English and Urdu, Critical Thinking, Positive Politics, Work Ethics and Professionalism, and Commercial awareness. Hence, the study produces implications for the employability stakeholders, including government and academia for a much needed shift from a mere subject-based curriculum to a skill-oriented curriculum and training in the Universities, particularly in the region of Southern Punjab, and all across Pakistan as well.

Innovation in Chinese internet companies: A meta-frontier analysis.

The innovation of a particular company benefits the whole industry when innovation technology transfers to others. Similarly, the development and innovation in internet companies influence the development and innovation of the industry. This investigation has applied a unique approach of meta-frontier analysis to estimate and analyze the innovation in internet companies in China. A unique dataset of Chinese internet companies from 2000 to 2017 has been utilized to estimate and compare the innovation over the period of study. The change in technology gap ratio (TGR) and a shift in production function have translated into innovation which was overlooked by previous studies. It is found that the production function of internet companies is moving upward in the presence of external factors such as smartphones invention, mobile internet, mobile payments, and artificial intelligence, etc. Consequently, a sudden increase in TGR is captured due to the innovation of some companies. Hence, the average TE of the industry falls caused by the increased distance of other companies form industry production function. However, the innovation advantage defused when other companies start imitating and the average TE elevates. A steady increase in the TGR index revealed that the continuous innovation-based growth of some companies lifting the production frontier upward. This provides the opportunity for other companies to imitate and provides continuous growth in the industry. This study provides a novel methodological approach to measure innovation and also provide practical implication by empirical estimation of innovation in Chinese internet companies.

Fighting Staphylococcus aureus Biofilms with Monoclonal Antibodies.

Staphylococcus aureus (S. aureus) is a notorious pathogen and one of the most frequent causes of biofilm-related infections. The treatment of S. aureus biofilms is hampered by the ability of the biofilm structure to shield bacteria from antibiotics as well as the host's immune system. Therefore, new preventive and/or therapeutic interventions, including the use of antibody-based approaches, are urgently required. In this review, we describe the mechanisms by which anti-S. aureus antibodies can help in combating biofilms, including an up-to-date overview of monoclonal antibodies currently in clinical trials. Moreover, we highlight ongoing efforts in passive vaccination against S. aureus biofilm infections, with special emphasis on promising targets, and finally indicate the direction into which future research could be heading.

Co-translational assembly of proteasome subunits in NOT1-containing assemblysomes.

The assembly of large multimeric complexes in the crowded cytoplasm is challenging. Here we reveal a mechanism that ensures accurate production of the yeast proteasome, involving ribosome pausing and co-translational assembly of Rpt1 and Rpt2. Interaction of nascent Rpt1 and Rpt2 then lifts ribosome pausing. We show that the N-terminal disordered domain of Rpt1 is required to ensure efficient ribosome pausing and association of nascent Rpt1 protein complexes into heavy particles, wherein the nascent protein complexes escape ribosome quality control. Immunofluorescence and in situ hybridization studies indicate that Rpt1- and Rpt2-encoding messenger RNAs co-localize in these particles that contain, and are dependent on, Not1, the scaffold of the Ccr4-Not complex. We refer to these particles as Not1-containing assemblysomes, as they are smaller than and distinct from other RNA granules such as stress granules and GW- or P-bodies. Synthesis of Rpt1 with ribosome pausing and Not1-containing assemblysome induction is conserved from yeast to human cells.