Structural and functional properties of inulinases: A review

Holyavka, M. G.; Artyukhov, V. G.; Kovaleva, T. A.

doi:10.1080/10242422.2016.1196486

Cited by 16 publications

(3 citation statements)

References 143 publications

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“…Unraveling the genome of the type strain ATCC 14580 T showed the presence of several genes encoding glycoside hydrolase enzymes putatively related to inulin hydrolysis: sacA , sacB , sacC , levB , and fruA . Most of these enzymes have been individually characterized, such as sucrose-6-phosphate hydrolase (β-fructofuranosidase, invertase) [ 25 ], levansucrase [ 26 , 27 , 28 , 29 ], levanase SacC [ 24 ], and endolevanase LevB [ 30 ]. A scheme of the glycosidic bonds that these enzymes usually target is shown in Figure 2 .…”

Section: Introductionmentioning

confidence: 99%

Influence of pH on Inulin Conversion to 2,3-Butanediol by Bacillus licheniformis 24: A Gene Expression Assay

Tsigoriyna,

Arsov,

Gergov

et al. 2023

IJMS

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2,3-Butanediol (2,3-BD) is an alcohol highly demanded in the chemical, pharmaceutical, and food industries. Its microbial production, safe non-pathogenic producer strains, and suitable substrates have been avidly sought in recent years. The present study investigated 2,3-BD synthesis by the GRAS Bacillus licheniformis 24 using chicory inulin as a cheap and renewable substrate. The process appears to be pH-dependent. At pH 5.25, the synthesis of 2,3-BD was barely detectable due to the lack of inulin hydrolysis. At pH 6.25, 2,3-BD concentration reached 67.5 g/L with rapid hydrolysis of the substrate but was accompanied by exopolysaccharide (EPS) synthesis. Since inulin conversion by bacteria is a complex process and begins with its hydrolysis, the question of the acting enzymes arose. Genome mining revealed that several glycoside hydrolase (GH) enzymes from different CAZy families are involved. Five genes encoding such enzymes in B. licheniformis 24 were amplified and sequenced: sacA, sacB, sacC, levB, and fruA. Real-time RT-PCR experiments showed that the process of inulin hydrolysis is regulated at the level of gene expression, as four genes were significantly overexpressed at pH 6.25. In contrast, the expression of levB remained at the same level at the different pH values at all-time points. It was concluded that the sacC and sacA/fruA genes are crucial for inulin hydrolysis. They encode exoinulinase (EC 3.2.1.80) and sucrases (EC 3.2.1.26), respectively. The striking overexpression of sacB under these conditions led to increased synthesis of EPS; therefore, the simultaneous production of 2,3-BD and EPS cannot be avoided.

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Section: Introductionmentioning

confidence: 99%

Influence of pH on Inulin Conversion to 2,3-Butanediol by Bacillus licheniformis 24: A Gene Expression Assay

Tsigoriyna,

Arsov,

Gergov

et al. 2023

IJMS

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“…Among them, the pair Gln-228/Asn-254, structurally equivalent to the Asp-239/Lys-242 pair of plant invertases 5 , was demonstrated by targeted mutation analysis to be significantly involved in the configuration of the acceptor-substrate binding site for transfructosylation and in the modulation of the enzymatic transfer specificity. Moreover, the Gln-176 residue linked to Arg-178 from the RDP motif, a specific Arg-Asp-Pro triad sequence evolutionarily preserved in β-fructofuranosidases and fructosyl-transferases for recognizing of the fructopyranosidic ring and stabilizing the transition state 6 , 7 , has proven to have a moderate influence on the substrate specificity and the catalytic efficiency of Ffase 5 . In addition to this, Ffase was found to have high fructosyl-acceptor promiscuity, being able to fructosylate 17 different hydroxylated compounds including small sugars and alditols, to produce new fructo-conjugates.…”

Section: Introductionmentioning

confidence: 99%

New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

Rodrigo-Frutos

Jiménez-Ortega

Piedrabuena

et al. 2021

Sci Rep

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The β-fructofuranosidase from Schwanniomyces occidentalis (Ffase) is a useful biotechnological tool for the fructosylation of different acceptors to produce fructooligosaccharides (FOS) and fructo-conjugates. In this work, the structural determinants of Ffase involved in the transfructosylating reaction of the alditols mannitol and erythritol have been studied in detail. Complexes with fructosyl-erythritol or sucrose were analyzed by crystallography and the effect of mutational changes in positions Gln-176, Gln-228, and Asn-254 studied to explore their role in modulating this biocatalytic process. Interestingly, N254T variant enhanced the wild-type protein production of fructosyl-erythritol and FOS by $$\sim$$ ∼ 30% and 48%, respectively. Moreover, it produced neokestose, which represented $$\sim$$ ∼ 27% of total FOS, and yielded 31.8 g l−1 blastose by using glucose as exclusive fructosyl-acceptor. Noteworthy, N254D and Q176E replacements turned the specificity of Ffase transferase activity towards the synthesis of the fructosylated polyols at the expense of FOS production, but without increasing the total reaction efficiency. The results presented here highlight the relevance of the pair Gln-228/Asn-254 for Ffase donor-sucrose binding and opens new windows of opportunity for optimizing the generation of fructosyl-derivatives by this enzyme enhancing its biotechnological applicability.

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“…Among them, the pair Gln-228/Asn-254 was demonstrated by targeted mutation analysis to be signi cantly involved in the con guration of the acceptor-substrate binding site for transfructosylation and in the modulation of the enzymatic transfer speci city. Moreover, the Gln-176 residue linked to Arg-178 from the RDP motif, a speci c Arg-Asp-Pro triad sequence evolutionarily preserved in β-fructofuranosidases and fructosyl-transferases for recognizing of the fructopyranosidic ring and stabilizing the transition state 6,7 , has proven to have a moderate in uence on the substrate speci city and the catalytic e ciency of Ffase 5 . In addition to this, Ffase was found to have high fructosyl-acceptor promiscuity, being able to fructosylate 17 different hydroxylated compounds including small sugars and alditols, to produce new fructo-conjugates.…”

Section: Introductionmentioning

confidence: 99%

New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

Rodrigo-Frutos

Jiménez-Ortega

Piedrabuena

et al. 2020

Preprint

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The β-fructofuranosidase from Schwanniomyces occidentalis (Ffase) is a useful biotechnological tool for the fructosylation of different acceptors to produce fructooligosaccharides (FOS) and fructo-conjugates. In this work, the structural determinants of Ffase involved in the transfructosylating reaction of the alditols mannitol and erythritol have been studied in detail. Complexes with fructosyl-erythritol or sucrose were analyzed by crystallography and the effect of mutational changes in positions Gln-176, Gln-228, and Asn-254 revised to explore their role in modulating this biocatalytic process. Interestingly, N254T variant enhanced the wild-type protein production of fructosyl-erythritol and FOS by `30% and 48%, respectively. Moreover, it showed a shift towards neokestose, which represented ~27% of total FOS, and yielded 31.8 g l-1 blastose by using glucose as exclusive fructosyl-acceptor. Noteworthy, N254D and Q176E replacements turned the specificity of Ffase transferase activity towards the synthesis of the fructosylated polyols at the expense of FOS production, but without increasing the total reaction efficiency. The results presented here highlight the relevance of the pair Gln-228/Asn-254 for Ffase donor-sucrose binding and opens new windows of opportunity for optimizing the generation of fructosyl-derivatives by this enzyme enhancing its biotechnological applicability.

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Structural and functional properties of inulinases: A review

Cited by 16 publications

References 143 publications

Influence of pH on Inulin Conversion to 2,3-Butanediol by Bacillus licheniformis 24: A Gene Expression Assay

Influence of pH on Inulin Conversion to 2,3-Butanediol by Bacillus licheniformis 24: A Gene Expression Assay

New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

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