Molecular and biochemical characteristics of inulosucrase InuBK from <i>Alkalihalobacillus krulwichiae</i> JCM 11691

Yokoi, Kohei; Tsutsui, Shigeyuki; Arakawa, Gen-ya; Takaba, Masakazu; Fujii, Koichi; Kaneko, Satoshi

doi:10.1093/bbb/zbab094

Cited by 8 publications

(1 citation statement)

References 36 publications

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“…8 In nature, inulins are synthesized by various plants (chicory, dahlia, dandelion, Jerusalem artichoke, etc. ), 10 lactic acid bacteria, 7 halophilic bacilli, 11 and halophilic Archaea. 12 Compared to the plantderived inulin, microbial inulin synthesized by ISs from sucrose generally shows higher molecular mass.…”

Section: ■ Introductionmentioning

confidence: 99%

Improving the Thermostability and Catalytic Activity of an Inulosucrase by Rational Engineering for the Biosynthesis of Microbial Inulin

Zhang

Kırtel

et al. 2021

J. Agric. Food Chem.

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Thermostability and enzymatic activity are two vital indexes determining the application of an enzyme on an industrial scale. A truncated inulosucrase, Laga-ISΔ138-702, from Lactobacillus gasseri showed high catalysis activity. To further enhance its thermostability and activity, multiple sequence alignment (MSA) and rational design based on the modeled structure were performed. Variants A446E, S482A, I614M, and A627S were identified with an improved denaturation temperature (T m) of more than 1 °C. A combinational mutation method was further carried out to explore the synergistic promotion effects of single-point mutants. Additionally, 33 residues at the N-terminus were truncated to construct mutant M4N‑33. The half-life of M4N‑33 at 55 °C increased by 120 times compared to that of Laga-ISΔ138-702, and the relative activity of M4N‑33 increased up to 152% at the optimal pH and temperature (pH 5.5 and 60 °C). Molecular dynamics (MD) simulations illustrated the decreased b-factor of the surface loop of M4N‑33.

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