Thermostable Acetaldehyde-Producing Enzymes: Characterization and Applications

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Microbiology".

Deadline for manuscript submissions: closed (15 June 2022) | Viewed by 5095

Special Issue Editor

Department of Biology, University of Waterloo, Waterloo, ON, Canada
Interests: enzymes; microbiology; biochemistry; enzymology; protein purification; protein analysis; hyperthermophiles
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Hyperthermophiles are a group of microorganisms growing optimally at ≥80 °C. They belong to the Bacteria or Archaea Domain and possess diverse metabolic capabilities and particularly novel thermostable enzymes. It has been found that many of them are involved in the production of aldehydes that are then reduced to alcohols at high temperatures. New types of acetaldehyde-producing enzymes are particularly of interest because of their important roles in biocatalysis and the production of bio-alcohols, such as ethanol, a most widely used biofuel. Pyruvate decarboxylase catalyzes the formation of acetaldehyde from pyruvate that is a central metabolic intermediate. Coenzyme A-dependent acetaldehyde dehydrogenase catalyzes the reduction of acetyl-CoA to acetaldehyde. Aldehyde ferredoxin oxidoreductase is involved in the formation of aldehydes from their corresponding carboxylic acids. Alcohol dehydrogenase catalyzes the reversible oxidation of alcohols to their corresponding aldehydes. These new thermostable enzymes show various biophysical and biochemical properties, such as the use of different cofactors and catalytic bifunctionalities. Further enhancement of their catalytic activities using bioengineering approaches will greatly promote their biotechnological applications in various industries. This Special Issue is dedicated to covering the topic of “Thermostable Acetaldehyde-Producing Enzymes: Characterization and Applications”.

Dr. Kesen Ma
Guest Editor

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Keywords

  • aldehyde
  • alcohol
  • pyruvate
  • decarboxylase
  • oxidoreductase
  • dehydrogenase
  • acetohydroxyacid synthase
  • hyperthermophile

Published Papers (2 papers)

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Research

9 pages, 1034 KiB  
Communication
Characterization of Thermotoga neapolitana Alcohol Dehydrogenases in the Ethanol Fermentation Pathway
by Chong Sha, Qiang Wang, Hongcheng Wang, Yilan Duan, Chongmao Xu, Lian Wu, Kesen Ma, Weilan Shao and Yu Jiang
Biology 2022, 11(9), 1318; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11091318 - 05 Sep 2022
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Abstract
Hyperthermophilic Thermotoga spp. are candidates for cellulosic ethanol fermentation. A bifunctional iron-acetaldehyde/alcohol dehydrogenase (Fe-AAdh) has been revealed to catalyze the acetyl-CoA (Ac-CoA) reduction to form ethanol via an acetaldehyde intermediate in Thermotoga neapolitana (T. neapolitana). In this organism, there are three [...] Read more.
Hyperthermophilic Thermotoga spp. are candidates for cellulosic ethanol fermentation. A bifunctional iron-acetaldehyde/alcohol dehydrogenase (Fe-AAdh) has been revealed to catalyze the acetyl-CoA (Ac-CoA) reduction to form ethanol via an acetaldehyde intermediate in Thermotoga neapolitana (T. neapolitana). In this organism, there are three additional alcohol dehydrogenases, Zn-Adh, Fe-Adh1, and Fe-Adh2, encoded by genes CTN_0257, CTN_1655, and CTN_1756, respectively. This paper reports the properties and functions of these enzymes in the fermentation pathway from Ac-CoA to ethanol. It was determined that Zn-Adh only exhibited activity when oxidizing ethanol to acetaldehyde, and no detectable activity for the reaction from acetaldehyde to ethanol. Fe-Adh1 had specific activities of approximately 0.7 and 0.4 U/mg for the forward and reverse reactions between acetaldehyde and ethanol at a pHopt of 8.5 and Topt of 95 °C. Catalyzing the reduction of acetaldehyde to produce ethanol, Fe-Adh2 exhibited the highest activity of approximately 3 U/mg at a pHopt of 7.0 and Topt of 85 °C, which were close to the optimal growth conditions. These results indicate that Fe-Adh2 and Zn-Adh are the main enzymes that catalyze ethanol formation and consumption in the hyperthermophilic bacterium, respectively. Full article
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14 pages, 2389 KiB  
Article
Thermostable and O2-Insensitive Pyruvate Decarboxylases from Thermoacidophilic Archaea Catalyzing the Production of Acetaldehyde
by Faisal Alharbi, Thomas Knura, Bettina Siebers and Kesen Ma
Biology 2022, 11(8), 1247; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11081247 - 22 Aug 2022
Cited by 1 | Viewed by 3254
Abstract
Pyruvate decarboxylase (PDC) is a key enzyme involved in ethanol fermentation, and it catalyzes the decarboxylation of pyruvate to acetaldehyde and CO2. Bifunctional PORs/PDCs that also have additional pyruvate:ferredoxin oxidoreductase (POR) activity are found in hyperthermophiles, and they are mostly oxygen-sensitive [...] Read more.
Pyruvate decarboxylase (PDC) is a key enzyme involved in ethanol fermentation, and it catalyzes the decarboxylation of pyruvate to acetaldehyde and CO2. Bifunctional PORs/PDCs that also have additional pyruvate:ferredoxin oxidoreductase (POR) activity are found in hyperthermophiles, and they are mostly oxygen-sensitive and CoA-dependent. Thermostable and oxygen-stable PDC activity is highly desirable for biotechnological applications. The enzymes from the thermoacidophiles Saccharolobus (formerly Sulfolobus) solfataricus (Ss, Topt = 80 °C) and Sulfolobus acidocaldarius (Sa, Topt = 80 °C) were purified and characterized, and their biophysical and biochemical properties were determined comparatively. Both enzymes were shown to be heterodimeric, and their two subunits were determined by SDS-PAGE to be 37 ± 3 kDa and 65 ± 2 kDa, respectively. The purified enzymes from S. solfataricus and S. acidocaldarius showed both PDC and POR activities which were CoA-dependent, and they were thermostable with half-life times of 2.9 ± 1 and 1.1 ± 1 h at 80 °C, respectively. There was no loss of activity in the presence of oxygen. Optimal pH values for their PDC and POR activity were determined to be 7.9 and 8.6, respectively. In conclusion, both thermostable SsPOR/PDC and SaPOR/PDC catalyze the CoA-dependent production of acetaldehyde from pyruvate in the presence of oxygen. Full article
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