Traditional screening methods of enzyme engineering often require building large mutant libraries to screen for potentially beneficial sites, which are often time-consuming and labor-intensive with low mining efficiency. In this study, a novel enzyme engineering strategy was established to modify carbonyl reductase LsCR for the synthesis of (1S)-2-chloro-1-(3,4-difluorophenyl) ethanol ((S)-CFPL), which is a key intermediate of anticoagulant drug ticagrelor. The strategy was developed by combining HotSpot, FireProt and multiple sequence alignment, resulting in the construction of a "small and smart" mutant library including 10 mutations. Among them, 5 mutations were positive, resulting in a 50% mining accuracy of beneficial sites. Finally, a highly active mutant LsCR (N101D/A117G/F147L) was obtained by further screening through saturation mutation and iterative mutation. Compared with wild type (WT) LsCR, the catalytic activity of LsCR was increased by 4.7 times, the catalytic efficiency k/K value was increased by 2.9 times, and the half-life t at 40 °C was increased by 1.3 times. Due to the low aqueous solubility of the substrate 2-chloro-1-(3,4-difluorophenyl) ethanone (CFPO), isopropanol was used as not only the co-substrate but also co-solvent. In the presence of 40% (v/v) isopropanol, LsCR completely reduced 400 g/L CFPO to enantiomerically pure CFPL (99.9%, e.e.) in 11 h with a space-time yield (STY) as high as 809 g/L∙d.
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http://dx.doi.org/10.1016/j.bioorg.2022.105991 | DOI Listing |
Scand J Gastroenterol
January 2025
Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan University, Xiamen Branch, Xiamen, China.
Background: Evaluate the clinical significance of esophagogastric junction (EGJ) morphology and esophagogastric junction contractile integral (EGJ-CI) in refractory gastroesophageal reflux disease (RGERD) patients.
Methods: From June 2021 to June 2023, 144 RGERD patients underwent comprehensive evaluation, recording symptom scores, demographic data. GERD classification (NERD or RE, A-D) was based on endoscopic findings.
Pharmacotherapy
January 2025
Department of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, Minnesota, USA.
Background: Lamotrigine clearance can change drastically in pregnant women with epilepsy (PWWE) making it difficult to assess the need for dosing adjustments. Our objective was to characterize lamotrigine pharmacokinetics in PWWE during pregnancy and postpartum along with a control group of nonpregnant women with epilepsy (NPWWE).
Methods: The Maternal Outcomes and Neurodevelopmental Effects of Antiepileptic Drugs (MONEAD) study was a prospective, observational, 20 site, cohort study conducted in the United States (December 2012 and February 2016).
Public Health Nutr
January 2025
Medical Research Council Research Unit for Hypertension and Cardiovascular Disease, North-West University, Potchefstroom, South Africa.
Objective: This study assessed the association between baseline sociodemographic variables, body composition and 4-year changes in the intake of food groups, including sugar sweetened beverages (SSBs) among children, after implementation of the health promotion levy.
Design: Prospective cohort study.
Setting: 10 schools in North West Province, South Africa.
Singapore Med J
January 2025
Department of Respiratory and Critical Care Medicine, Singapore General Hospital, Singapore.
Introduction: Rapid response teams (RRTs) are prevalent in healthcare institutions worldwide. Repeated activations are associated with increased morbidity and higher resource utilisation, and represent a heterogeneous population that may benefit from early identification. To date, there are no published data on repeat RRT activations in Singapore.
View Article and Find Full Text PDFBiosci Microbiota Food Health
September 2024
Core Technology Laboratories, Asahi Quality & Innovations, Ltd., 1-1-21 Midori, Moriya-shi, Ibaraki 302-0106, Japan.
α-Cyclodextrin (αCD), a cyclic hexasaccharide composed of six glucose units, is not digested in the small intestine but is completely fermented by gut microbes. Recently, we have reported that αCD supplementation for nonathlete men improved their 10 km biking times. However, the beneficial effects of αCD on exercise are not yet fully understood.
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