Sangyoung Yoon

Characterization of a novel oligomeric SGNH-arylesterase from Sinorhizobium meliloti 1021.

A novel oligomeric SGNH-arylesterase (Sm23) from Sinorhizobium meliloti 1021 was characterized using biochemical and biophysical methods. A sequence comparison of Sm23 with other SGNH members confirmed the presence of catalytic triad (Ser(10), Asp(187), and His(190)) and oxyanion holes (Ser(10)-Gly(50)-Asn(90)). The wild type enzyme was able to hydrolyze p-nitrophenyl acetate, alpha- and beta-naphthyl acetate, while S10A mutant completely lost its activity. Structural properties of Sm23 were investigated using circular dichroism (CD), fluorescence, dynamic light scattering (DLS), chemical cross-linking, electron microscopy (EM), and time of flight (TOF) mass spectrometry. Furthermore, spherical or globular aggregates were observed with 1-butyl-3-methylimidazolium tetrafluoroborate, while amorphous aggregates were formed with 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide.

Improved enantioselectivity of thermostable esterase from Archaeoglobus fulgidus toward (S)-ketoprofen ethyl ester by directed evolution and characterization of mutant esterases

Thermostable esterases have potential applications in various biotechnology industries because of their resistance to high temperature and organic solvents. In a previous study, we isolated an esterase from Archaeoglobus fulgidus DSM 4304 (Est-AF), which showed high thermostability but low enantioselectivity toward (S)-ketoprofen ethyl ester. (R)-ketoprofenor (S)-ketoprofenis produced by esterase hydrolysis of the ester bond of (R,S)-ketoprofen ethyl ester and (S)-ketoprofen has better pharmaceutical activity and lower side effects than (R)-ketoprofen. Therefore, we have generated mutants of Est-AF that retained high thermostability whilst improving enantioselectivity. A library of Est-AF mutants was created by error-prone polymerase chain reaction, and mutants with improved enantioselectivity were isolated by site-saturation mutagenesis. The regions of Est-AF containing amino acid mutations were analyzed by homology modeling of its three-dimensional structure, and structure-based explanations for the changes in enantioselectivity are proposed. Finally, we isolated two mutants showing improved enantioselectivity over Est-AF (ee% = −16.2 ± 0.2 and E = 0.7 ± 0.0): V138G (ee% = 35.9 ± 1.0 and E = 3.0 ± 0.1) and V138G/L200R (ee% = 89.2 ± 0.2 and E = 19.5 ± 0.5). We also investigated various characteristics of these mutants and found that the mutants showed similar thermostability and resistance to additives or organic solvents to Est-AF, without a significant trade-off between activity and stability.

The Survey of Job Rotation Implementation at Medium- and Small-Industries

Objective: The aim of this study is to investigate the job rotation implementation at medium- or small-industries and to identify the viewpoint on job rotation as preventive activity of work-related musculoskeletal disorders(WMSDs). Background: Job rotation has been implemented in many industrial areas in order to prevent the WMSDs as one part of ergonomic program. Generally, the cases of implementation of ergonomic program or successful cases of job rotation were reported on the side of major or large company. Therefore, this study tried to inspect the current state of job rotation implementation at medium- or small-industries. Method: Survey was carried out for randomly contacted forty seven mangers responsible for safety. Survey contained the questionnaires on the general state of company, shift-work and job rotation. Results: The ratio of work-shift in medium- and small-industry was 34.0% and the ratio of job rotation was 19.1%. For manufacturing industry, the ratio was 37.9% and 17.2%, respectively. Conclusion: The implementation ratio of job rotation was relatively low considering the results of previous studies. Many managers appealed the quality decreasing of goods and the injuries of workers due to job rotation, though agreed to train the multi-functional worker and to prevent the WMSDs. Application: The results can be used for the fundamental data how the job rotation will be properly implemented in medium- and small-industry as an administrative control for MSDs.

Purification, crystallization and preliminary crystallographic analysis of Est-Y29: a novel oligomeric β-lactamase

beta-Lactam antibiotics such as penicillins and cephalosporins have a four-atom ring as a common element in their structure. The beta-lactamases, which catalyze the inactivation of these antibiotics, are of great interest because of their high incidence in pathogenic bacteria. A novel oligomeric class C beta-lactamase (Est-Y29) from a metagenomic library was expressed, purified and crystallized. The recombinant protein was expressed in Escherichia coli with an N-terminal 6xHis tag and purified to homogeneity. EstY-29 was crystallized and X-ray intensity data were collected to 1.49 A resolution using synchrotron radiation.

Hands-on Tools to Prevent Human Errors in Highway Construction

Objective: The aim of this study is to reclassify human errors and to develop hands-on tools to apply the new classification for preventing human error accidents in highway construction site. Background: The main cause of accidents in highway construction was reported as the carelessness of workers. However, such diagnosis could not help us operationally prevent accidents in real workplace. Method: The accidents in highway construction were reanalyzed and the causes of human error were reclassified in order to educate and improve the awareness of human error in highway construction. Field survey and interview with safety managers and workers were conducted to find the causal relationship between the actual accidents and the human errors. Results: The most frequently observed human errors in highway construction were classified into six categories such as mis-perception, distraction, memory fail, slip, cognition error and mis-judgment. In order to provide hands-on tools to increase the awareness of human error in construction field, the human error checklist and card sorting diary were developed. Especially, the card sorting diary was designed to increase the ability in human error inspection of safety manager at construction site. Moreover, posters were developed based on actual accident cases. Conclusion: We suggested that the improved awareness and analytical report on checklist, card sorting diary and posters for construction field could collectively prevent the accident. Application: The classification of human error, hands-on tools and posters can be directly applicable on highway construction site. This analytical and collective approach preventing human error-related accident could be extended to other construction workplaces.

Identification and Characterization of a Novel Oligomeric Decaprenyl Diphosphate Synthase

A novel oligomeric decaprenyl pyrophosphate synthase was identified. Circular dichroism measurements indicated that it is alpha-helical, and stable against pH changes and denaturants. Three peptides corresponding to the conserved regions were synthesized and metal affinities were investigated. Crystallographic analyses of this oligomeric DDS are currently in progress.

Trends of Intellectual Property on Musculoskeletal Disorder, Motion Capture Technology and Ergonomics

Objective:The aims of this study are to investigate the trends of intellectual property in order to identify the ergonomic approaches on musculoskeletal disorders, harmful factors of musculoskeletal disorders, and to find the potential applicability of motion capture technology. Background: Ergonomic posture assessment tools often showed interrater variance, though the usage is easy and practical in industrial fields. Moreover new technologies such as motion capture showed the potential applicability in posture assessment. So ergonomists and practitioners became interested in the intellectual properties on musculoskeletal disorder and motion capture technology. Method: Intellectual properties were collected with the combination of keywords such as ergonomic, musculoskeletal disorder, and motion capture using the KIPRIS(Korea Intellectual Property Rights Information Service). Collected intellectual properties were classified into ergonomic area and non-ergonomic area, except unexamined intellectual properties. This study investigated the trend of application of intellectual properties and the probability of using motion capture technology. Results: Few intellectual properties with ergonomic approach on musculoskeletal disorders were founded, despite many products for rehabilitation and sports. One hundred twenty five patents in 1105 patents on musculoskeletal disorders and 138 patents in 1908 patents on motion capture technology were classified into the patents that ergonomic approach can be applied. The patents related to ergonomics area are rapidly increasing after 2010, and there are good opportunities for ergonomists to apply the patents. Conclusion: This study found opportunities on novel methodology in detecting the harmful factors of musculoskeletal disorders, and that the motion capture technology is applicable in ergonomic posture assessment. Application: The results of this study can help ergonomists prepare the ergonomic patents, and can show the potential use of motion capture technology in detecting the harmful posture of musculoskeletal disorders.

Literature Review on Job Rotation

Objective: The aim of this study is to review the studies on job rotation with various points of view and to classify the studies considering the approach methods. Background: Various work interventions were carried in order to prevent the work-related musculoskeletal disorders(WMSDs) and job rotation was considered as one of the simple and effective methods. Method: Previous studies were systematically collected with the combination of keywords such as job, task, rotation, allocation, line balancing, etc. Collected studies were classified into three categories: field study, laboratory study and optimization approach study. Results: Field studies showed that most studies focused on the evaluation of the factors causing the musculoskeletal disorder. Moreover, the effect of job rotation could be hard to identify because it required a long-period close observation. Laboratory studies covered only simple aspects of tasks or movements. Optimization approach studies showed the systematic approach to establish the job rotation scheme though many limitation and assumptions were adopted. Conclusion: This study collected and classified the previous studies on job rotation and presented the different approaches across various research areas. Application: The results of this study could help the practitioner and ergonomist how to establish the job rotation scheme.