Identification and expression of a hydrophobic carboxylic acid reductase from Trichoderma virens

被引:0
|
作者
Goh, Andrew B. H. [1 ]
Ling, Jonathan G. [1 ]
Kamaruddin, Shazilah [1 ]
Murad, Abdul M. A. [1 ]
Abu Bakar, Farah D. [1 ]
机构
[1] Univ Kebangsaan Malaysia, Fac Sci & Technol, Dept Biol Sci & Biotechnol, Ukm Bangi 43600, Selangor, Malaysia
关键词
oxidoreductase; hydrophobic enzyme; NRPS-like enzyme; protein expression; protein solubility; carboxylic acid reductase; BIOSYNTHESIS; DISCOVERY; NRPS; ENZYMES; CLUSTER; DOMAIN;
D O I
10.1093/femsle/fnaf021
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Carboxylic acid reductases (CARs) have been garnering attention in applications for the sustainable synthesis of aldehydes. Despite numerous discoveries, not all characteristics of CAR enzymes have been extensively studied or understood. Herein, we report the discovery and expression of a new CAR enzyme (TvirCAR2) from the ascomycetous fungus, Trichoderma virens. Tvircar2 is one of the five putative CARs identified from analyses of the T. virens genome. In silico, analyses showed that TvirCAR2 has a high hydrophobicity index and that its corresponding gene is part of a biosynthetic gene cluster predicted to synthesize hybrid polyketide synthases-nonribosomal peptide synthetase secondary metabolites. TvirCAR2 was highly expressed as soluble and insoluble forms in an Escherichia coli expression host. The solubility of the purified TvirCAR2 necessitated the addition of glycerol in the purification and assay buffers. Substrate screening via molecular docking showed that benzoic acid was a suitable substrate candidate. The TvirCAR2 enzyme catalyzed the reduction of benzoic acid with a specific activity of around 1.4 mu mol/h/mg. Homologs, which are predicted to exhibit similar hydrophobicity, are the CARs from Stachybotrys bisbyi (StbB), which is involved in the production of the meroterpenoid, ilicicolin B, and Trichoderma reesei (TrCAR), which is part of a similar but still uncharacterized biosynthetic gene cluster. We have identified, produced and characterized a new novel carboxylic acid reductase (CAR) from the filamentous fungus Trichoderma virens. We have identified features not previously seen or reported in other CARs, which supports the role of this CAR as a natural tailoring enzyme in a hybrid polyketide synthase/ nonribosomal peptide synthetase-linked pathway.
引用
收藏
页数:13
相关论文
共 50 条
  • [31] Isolation and cultivation of xylanolytic and cellulolytic Sarocladium kiliense and Trichoderma virens from the gut of the termite Reticulitermes santonensis
    Tarayre, Cedric
    Bauwens, Julien
    Brasseur, Catherine
    Matteotti, Christel
    Millet, Catherine
    Guiot, Pierre Alexandre
    Destain, Jacqueline
    Vandenbol, Micheline
    Portetelle, Daniel
    De Pauw, Edwin
    Haubruge, Eric
    Francis, Frederic
    Thonart, Philippe
    ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH, 2015, 22 (06) : 4369 - 4382
  • [32] An engineered fatty acid synthase combined with a carboxylic acid reductase enables de novo production of 1-octanol in Saccharomyces cerevisiae
    Henritzi, Sandra
    Fischer, Manuel
    Grininger, Martin
    Oreb, Mislav
    Boles, Eckhard
    BIOTECHNOLOGY FOR BIOFUELS, 2018, 11
  • [33] PURIFICATION AND SOME PROPERTIES OF THE TUNGSTEN-CONTAINING CARBOXYLIC-ACID REDUCTASE FROM CLOSTRIDIUM-FORMICOACETICUM
    WHITE, H
    FEICHT, R
    HUBER, C
    LOTTSPEICH, F
    SIMON, H
    BIOLOGICAL CHEMISTRY HOPPE-SEYLER, 1991, 372 (11): : 999 - 1005
  • [34] Gliotoxin, an Immunosuppressive Fungal Metabolite, Primes Plant Immunity: Evidence from Trichoderma virens-Tomato Interaction
    Zaid, Rinat
    Koren, Roni
    Kligun, Efrat
    Gupta, Rupali
    Leibman-Markus, Meirav
    Mukherjee, Prasun K.
    Kenerley, Charles M.
    Bar, Maya
    Horwitz, Benjamin A.
    MBIO, 2022, 13 (04):
  • [35] In VivoReduction of Medium- to Long-Chain Fatty Acids by Carboxylic Acid Reductase (CAR) Enzymes: Limitations and Solutions
    Horvat, Melissa
    Winkler, Margit
    CHEMCATCHEM, 2020, 12 (20) : 5076 - 5090
  • [36] Engineering Carboxylic Acid Reductase (CAR) through a Whole-Cell Growth-Coupled NADPH Recycling Strategy
    Kramer, Levi
    Le, Xuan
    Rodriguez, Marisa
    Wilson, Mark A.
    Guo, Jiantao
    Niu, Wei
    ACS SYNTHETIC BIOLOGY, 2020, 9 (07): : 1632 - 1637
  • [37] Carboxylic acid reductase is a versatile enzyme for the conversion of fatty acids into fuels and chemical commodities
    Akhtar, M. Kalim
    Turner, Nicholas J.
    Jones, Patrik R.
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2013, 110 (01) : 87 - 92
  • [38] Glutathione Transferase from Trichoderma virens Enhances Cadmium Tolerance without Enhancing Its Accumulation in Transgenic Nicotiana tabacum
    Dixit, Prachy
    Mukherjee, Prasun K.
    Ramachandran, V.
    Eapen, Susan
    PLOS ONE, 2011, 6 (01):
  • [39] Production of α-keto carboxylic acid dimers in yeast by overexpression of NRPS-like genes from Aspergillus terreus
    Huehner, Elisabeth
    Backhaus, Katja
    Kraut, Rixa
    Li, Shu-Ming
    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2018, 102 (04) : 1663 - 1672
  • [40] Control of parallelized bioreactors II: probabilistic quantification of carboxylic acid reductase activity for bioprocess optimization
    Nikolas von den Eichen
    Michael Osthege
    Michaela Dölle
    Lukas Bromig
    Wolfgang Wiechert
    Marco Oldiges
    Dirk Weuster-Botz
    Bioprocess and Biosystems Engineering, 2022, 45 : 1939 - 1954