A highly thermostable lichenase from Bacillus sp. UEB-S: Biochemical and molecular characterization

Maktouf, Sameh; Moulis, Claire; Miled, Nabil; Châabouni, Semia Ellouz; Remaud‐Siméon, Magali

doi:10.1016/j.molcatb.2015.01.016

Cited by 6 publications

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References 29 publications

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“…3.2.1.73) has received notable attention, thanks to its significant role in exploring the feasibility of biofuel production [ 1 ], though some thermostable lichenases from various origins have been isolated, for example: Bacillus sp. UEB-S [ 2 ], Aspergillus niger US368 [ 3 ] and Clostridium thermocellum [ 4 ]. However, current commercially available lichenases may not be ideally suitable for biomass biocatalytic conversion because of its limited thermal stability.…”

Section: Introductionmentioning

confidence: 99%

Enhanced thermal stability of lichenase from Bacillus subtilis 168 by SpyTag/SpyCatcher-mediated spontaneous cyclization

Wang

et al. 2016

Biotechnol Biofuels

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BackgroundSpyTag is a peptide that can form an irreversible covalent linkage to its 12 kDa partner SpyCatcher via a spontaneous isopeptide bond. Herein, we fused SpyTag at the N-terminal of lichenase and SpyCatcher at C-terminal so that the termini of lichenase were locked together by the covalent interaction between the partners. In addition, an elastin-like polypeptides tag was subsequently attached to the C-terminus of SpyCatcher, thereby facilitating the non-chromatographic purification of cyclized lichenase.ResultsThe study showed that the optimum temperature of the cyclized lichenase was about 5 °C higher in comparison to its linear counterpart. Moreover, nearly 80 % of the cyclized lichenase activities were retained after 100 °C exposure, whereas the linear form lost almost all of its activities. Therefore, the cyclized variant displayed a significantly higher thermal stability as temperature elevated and was resistant to hyperthermal denaturation. Besides, the Km value of the cyclized lichenase (7.58 ± 0.92 mg/mL) was approximately 1.7-fold lower than that of the linear one (12.96 ± 1.93 mg/mL), indicating a higher affinity with substrates.ConclusionsThis new SpyTag/SpyCatcher cyclization strategy is deemed as a generalized reference for enhancing enzyme stability and can be effectively customized to the cyclization of various enzymes, hence a tremendous potential for successful application in the biocatalytic conversion of biomass to produce fuels and chemicals.

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