The Thermoanaerobacter ethanolicus secondary alcohol dehydrogenase I86A mutant is stereospecific for (R)-alcohols instead of (S)-alcohols. Pyramidal crystals grown in the presence of (R)-phenylethanol via the hanging-drop vapour-diffusion method diffracted to 3.2 A resolution at the Canadian Light Source. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 80.23, b = 124.90, c = 164.80 A. The structure was solved by molecular replacement using the structure of T. brockii SADH (PDB entry 1ykf).
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http://dx.doi.org/10.1107/S1744309110018981 | DOI Listing |
Appl Environ Microbiol
June 2023
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
Genetic engineering of hyperthermophilic organisms for the production of fuels and other useful chemicals is an emerging biotechnological opportunity. In particular, for volatile organic compounds such as ethanol, fermentation at high temperatures could allow for straightforward separation by direct distillation. Currently, the upper growth temperature limit for native ethanol producers is 72°C in the bacterium Thermoanaerobacter ethanolicus JW200, and the highest temperature for heterologously-engineered bioethanol production was recently demonstrated at 85°C in the archaeon Pyrococcus furiosus.
View Article and Find Full Text PDFChem Commun (Camb)
October 2022
Department of Chemistry, University of Oxford, South Parks Road, Oxford, OX1 3QR, UK.
The unique ability of the 'electrochemical leaf' (e-Leaf) to drive and control nanoconfined enzyme cascades bidirectionally, while directly monitoring their rate in real-time as electrical current, is exploited to achieve deracemisation and stereoinversion of secondary alcohols using a single electrode in one pot. Two alcohol dehydrogenase enzymes with opposing enantioselectivities, from (selective for ) and (selective for ) are driven bidirectionally coupling to the fast and quasi-reversible interconversion of NADP/NADPH catalysed by ferredoxin NADP reductase - all enzymes being co-entrapped in a nanoporous indium tin oxide electrode. Activity of the enzyme depends on the binding of a non-catalytic Mg, allowing it to be switched off after an oxidative half-cycle, by adding EDTA - the -selective enzyme, with a tightly-bound Zn, remaining fully active.
View Article and Find Full Text PDFBiotechnol Lett
October 2022
Centro de Estudios Fotosintéticos y Bioquímicos (CEFOBI), CONICET-Universidad Nacional de Rosario, Suipacha 531, 2000, Rosario, Argentina.
Purpose: We identified a new glucoamylase (TeGA) from Thermoanaerobacter ethanolicus, a thermophilic anaerobic bacterium. Structural studies suggest that TeGA belongs to the family 15 of glycosylhydrolases (GH15).
Methods: The expression of this enzyme was optimized in E.
Enzyme Microb Technol
December 2021
The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, PR China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, PR China; Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, PR China.
Appl Environ Microbiol
December 2020
Biofuels Institute, School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang, Jiangsu, People's Republic of China
can produce acetate, lactate, hydrogen, and ethanol from sugars resulting from plant carbohydrate polymer degradation at temperatures above 65°C. is a promising candidate for thermophilic ethanol fermentations due to the utilization of both pentose and hexose. Although an ethanol balance model in has been developed, only a few physiological or biochemical experiments regarding the function of important enzymes in ethanol formation have been carried out.
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