Dataset Information

A hydrolase with esterase activity expressed from a fosmid gene bank prepared from DNA of a North West Himalayan glacier frozen soil sample.

ABSTRACT: Screening of 20,000 clones of a fosmid gene bank, constructed from DNA extracted from North West Himalaya (NWH) glacier soil sample, using functional approach identified 10 esterase/lipase-producing clones. Of these, a clone designated pFG43 with an insert size of 45 kb which produced the highest concentration of enzyme (467.43 U/mg) was sequenced. Clone pFG43 contained 61 open reading frames (ORF) and of these an ORF of 1155 bp designated ME-003, was found to be closely related to a hydrolase from Acidobacteria sps (77% sequence identity and E value?=?1e-164) and subsequently identified as a putative cocaine esterase. ORF ME-003 was amplified and sub-cloned using a TA vector system into E. coli (DH5?). The purified recombinant enzyme with a molecular weight of 43 kDa had optimal activity at 40 °C, pH 6 and the highest activity with shorter chain fatty acids than with higher chain length fatty acids. There is insignificant effect of inhibitors on the enzyme activity of ME-003, except PMSF which completely inhibited its activity. ME-003 activity was also inhibited in the presence of copper oxide but remained stable in presence of other metal ions. The enzyme activity was also inhibited in the presence of organic solvents; however, in the presence of 10% isopropanol, 12% of enzymatic activity was retained. Among various detergents, SDS completely inhibited enzymatic activity. The recombinant enzyme also shows enantio-specific activity against the racemic drug intermediates/precursors and exhibited 90% ee against racemic 1-phenyl ethanol and fluoxetine.

SUBMITTER: Gupta V

PROVIDER: S-EPMC6391513 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

A hydrolase with esterase activity expressed from a fosmid gene bank prepared from DNA of a North West Himalayan glacier frozen soil sample.

Gupta Verruchi V Singh Inderpal I Kumar Paramdeep P Rasool Shafaq S Verma Vijeshwar V

3 Biotech 20190226 3

Screening of 20,000 clones of a fosmid gene bank, constructed from DNA extracted from North West Himalaya (NWH) glacier soil sample, using functional approach identified 10 esterase/lipase-producing clones. Of these, a clone designated pFG43 with an insert size of 45 kb which produced the highest concentration of enzyme (467.43 U/mg) was sequenced. Clone pFG43 contained 61 open reading frames (ORF) and of these an ORF of 1155 bp designated ME-003, was found to be closely related to a hydrolase f ...[more]

PMID: 30863691

Similar Datasets

Project description:BackgroundIn today's companies, time management abilities have grown as a significant predictor of nurses' success. Organizations have simplified their internal operations and flattened their organizational structures in an effort to increase productivity and cut expenses. As a result, successful time management skills are particularly crucial for nurses in recently restructured healthcare organizations. This study aimed at exploring factors influencing time management skills among Palestinian nurses.MethodsCross-sectional quantitative study of all nurses (715) working in private and government hospitals and primary healthcare centers in north Palestine was conducted. Time management skills were measured on a continuous scale using the Nursing Time Management Scale (NTMS), Arabic version. The scale measures various aspects of time management including goal setting, planning, scheduling, and organizing activities. The relationship between time management skills and background variables was assessed using the multivariate linear regression.ResultsThe average total score for NTMS scale was 63.39 out of a total score of 90. This score indicates relatively good time management skills among the respondents. The multivariate linear regression results showed that females obtained slightly lower scores than males, coefficient = -2.36, p = 0.043. Nurses in primary care centers had significantly higher scores than nurses who work at hospitals, coefficient = 4.47, p = 0.004. The type of healthcare organization emerged as a significant factor predicting time management skills. Nurses in private hospitals had worse time management skills than nurses in government hospitals, coefficient = -12.27, p < 0.001. Nurse supervisors had better time management skills than staff nurses, coefficient = 4.01, p = 0.023. Nurses working in non-teaching hospitals had worse time management skills than nurses in teaching hospitals, coefficient = - 3.86, p = 0.001. Nurses who did not attend a time management course had worse time management skills than nurses who attended time management course, coefficient = - 4.05, p = p < 0.001.ConclusionsHealthcare institutions should consider organizational and individual factors to improve the time management skills of their staff. Time management training interventions are proven and effective policies that are recommended to be adopted by all healthcare centers.

Dataset Information

A hydrolase with esterase activity expressed from a fosmid gene bank prepared from DNA of a North West Himalayan glacier frozen soil sample.

Publications

A hydrolase with esterase activity expressed from a fosmid gene bank prepared from DNA of a North West Himalayan glacier frozen soil sample.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets