Cloning, expression, and biochemical characterisation of a novel endomannanase from Thermobifida alba
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Published:2023-09-11
Issue:3
Volume:52
Page:502-519
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ISSN:0139-3006
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Container-title:Acta Alimentaria
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language:
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Short-container-title:AAlim
Author:
Luzics Sz.1, Tóth Á.2, Barna T.3, Szabó E.3, Nagy I.1, Horváth B.4, Nagy I.4, Varecza Z.5, Bata-Vidács I.1ORCID, Kukolya J.1
Affiliation:
1. Eszterházy Károly Catholic University, Food and Wine Research Institute, Eger, Hungary 2. Heart and Vascular Center, Semmelweis University, Budapest, Hungary 3. Department of Genetics and Applied Microbiology, University of Debrecen, Hungary 4. Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary 5. eXom Ltd., Pécs, Hungary
Abstract
AbstractThermobifida alba is the mesophilic member of the Thermobifida genus, the genome and enzyme sets of which have not been described and published yet. Thermobifida strains are thermotolerant actinomycete, which possess wide sets of cellulose and hemicellulose hydrolysing enzymes. Previously, three endomannanases (Man5ATh, Man5ATc, and Man5AThf) of thermobifidas were cloned and investigated, and hereby the endomannanase of T. alba DSM 43795 is described. All four endomannanases belong to the glycoside hydrolase family 5, their sizes are around 50–55 kDa. Their structure consists of a catalytic domain and a carbohydrate binding module, while there is an interdomain linker region in-between consisting repetitive tetrapeptide motifs (eg.: PPTEPTD-Ta, PTDP-Tc, TEEP-Tf, DPGT-Th). The pH optima of Man5A enzymes from T. alba, Thermobifida halotolerans, Thermobifida cellulosilytica, and Thermobifida fusca are slightly different (6.5, 7.0, 7.5, and 8.0, respectively), however, the temperature optima of the enzymes were detected within a wider range of 65–75 °C. In this research, Man5ATa exhibited the lowest Michaelis-Menten constant (KM) (0.13 mM) on LBG-mannan substrate, while others shared similar kinetic parameters: 0.9–1.7 mM of KM. Despite the high sequence similarity of the investigated mannanases, they exhibit different temperature stability parameters. These different functional characteristics can be advantageous for industrial applications producing biologically active, oligomannan prebiotics under different conditions.
Funder
Hungarian Academy of Sciences
Publisher
Akademiai Kiado Zrt.
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