Molecular cloning and anti-fungal effect of endo-β-1,3-glucanase from Thermotoga maritima

Woo, Chang Bin; Kang, Hae Na; Lee, Soo Bok

doi:10.1007/s10068-014-0170-9

Cited by 11 publications

(7 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…β-1,3-Glucanase displayed high tolerance towards DMSO, with >70% activity retained, while acetonitrile and acetone significantly decreased enzyme activity up to 50% (Figure 5). These results are concurrent with those of previous studies reporting that methanol, ethanol, and DMSO (all 10%) decreased the relative activity of Thermotoga maritima β-1,3-glucanase by 52%, 34% and 16%, respectively [35].…”

Section: Characterization Of β-13-glucanase From Y Lipolytica Transformant (Pylsc1-13g)supporting

confidence: 91%

Expression of the Thermobifida fusca β-1,3-Glucanase in Yarrowia lipolytica and Its Application in Hydrolysis of β-1,3-Glucan from Four Kinds of Polyporaceae

et al. 2020

View full text Add to dashboard Cite

The gene encoding a thermostable β-1,3-glucanase was cloned from Thermobifida fusca and expressed constitutively by Yarrowia lipolytica using plasmid pYLSC1. The expression level of the recombinant β-1,3-glucanase reached up to 270 U/mL in the culture medium. After a treatment with endo-β-N-acetyl-glucosaminidase H, the recombinant protein appeared as a single protein band, with a molecular size of approximately 66 kDa on the SDS-polyacrylamide gel. The molecular weight was consistent with the size predicted from the nucleotide sequence. The optimum temperature and pH of the transformant β-1,3-glucanase were 60 °C and pH 8.0, respectively. This β-1,3-glucanase was tolerant to 10% methanol, ethanol, and DMSO, retaining 70% activity. The enzyme markedly hydrolyzed Wolfiporia cocos and Pycnoporus sanguineus glucans. The DPPH and ABTS scavenging potential, reducing power and total phenolic contents of these two Polyporaceae hydrolysates, were significantly increased after 18 h of the enzymatic reaction. The present results indicate that T. fusca β-1,3-glucanase from Y. lipolytica transformant (pYLSC1-13g) hydrolyzes W. cocos and P. sanguineus glucans and improves the antioxidant potential of the hydrolysates.

show abstract

Section: Characterization Of β-13-glucanase From Y Lipolytica Transformant (Pylsc1-13g)supporting

confidence: 91%

Expression of the Thermobifida fusca β-1,3-Glucanase in Yarrowia lipolytica and Its Application in Hydrolysis of β-1,3-Glucan from Four Kinds of Polyporaceae

et al. 2020

View full text Add to dashboard Cite

show abstract

“…In particular, as a result of barley β-glucan hydrolises, Jermuk-LamM produces minimal glucose, while FLamA and FLamB generate glucose as one of the primary end products [89]. Thus, based on the biochemical characteristics, substrate specificity and product pattern we classify Jermuk-LamM as a typical endo-β-1,3-glucanase of the GH16_3 subfamily (EC 3.2.1.39) [90] together with TmβG from the bacterium Thermotoga maritima MSB8 [43] and LamA from the archaeon Pyrococcus furiosus , respectively [91].…”

Section: Discussionmentioning

confidence: 99%

“…Laminarinases have been characterised from plants [28], fungi [29,30,31,32], algae [33], Actinobacteria [34,35], Archaea [36], soil bacteria [37,38,39], thermophilic [40,36,41,42,43,44] and marine bacteria [45,46,47,21,48,49]. Most of the characterised laminarinases belong to glycosyl hydrolase family 16 (GHF16) [50].…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning and Biochemical Characterisation of a Novel Acidic Laminarinase Derived from Jermuk Hot Spring Metagenome

Paloyan,

Karapetyan,

Grigoryan

et al. 2024

Preprint

View full text Add to dashboard Cite

Laminarinase, an enzyme with a specific affinity for laminarin, a complex polysaccharide found in the cell walls of brown algae and select marine organisms, was investigated in this study. We cloned and characterised a gene encoding a putative glycoside hydrolase family 16 (GH16) laminarinase from the Jermuk hot spring metagenome by heterologous expression in Escherichia coli. The resulting product, named Jermuk-LamM, represents a novel endo-1,3-beta-D-glucanase (EC 3.2.1.39) with only 48.1 % amino acid sequence similarity to previously characterised GH16 family members catalogued in the NCBI database. To date, this stands as the sole described endo-1,3-beta-D-glucanase within the Marinimicrobia phylum. Jermuk-LamM, identified as an acidic laminarinase, exhibits robust enzymatic activity at pH 5.0 and a temperature of 55 0C, maintaining its function for a duration of at least 7 hours. Notably, this enzyme effectively catalyses the hydrolysis of both soluble and insoluble (1,3)-beta-D-glucans, as well as (1,3;1,4)-beta-D-glucans, displaying a pronounced preference for laminarin. The specificity of Jermuk-LamM lies in its cleavage of 1,3-beta-D-glucosidic linkages, yielding monosaccharides, disaccharides, and oligosaccharides. These breakdown products hold the potential for conversion into energy carriers, including alcohols, methane, and hydrogen. The enzyme's exceptional specific activities, coupled with its resistance to various additives, render Jermuk-LamM a promising candidate for various industrial applications, encompassing the realms of biofuel and pharmaceutical production.

show abstract

“…These results indicated that the anti-fungal activity of rGluY against C. albicans was relatively stronger than those of other known functional homologs against the same fungal pathogen. Previously, S. matensis ATCC 23,935 GH64 endo-β-1,3-glucanase (SmβG) [8] and Thermotoga marimata endo-β-1,3-glucanase (TmβG) [34] were also reported to inhibit over…”

Section: Anti-fungal Activities Of Rgluy and Rgluyδricinmentioning

confidence: 99%

“…These results indi-cated that the anti-fungal activity of rGluY against C. albicans was relatively stronger than those of other known functional homologs against the same fungal pathogen. Previously, S. matensis ATCC 23,935 GH64 endo-β-1,3-glucanase (SmβG) [8] and Thermotoga marimata endo-β-1,3-glucanase (TmβG) [34] were also reported to inhibit over 60% of the growth of C. albicans at concentrations exceeding 0.1 mg/mL (approximately 2 µM) and 0.2 mg/mL (approximately 2.7 µM), respectively. Compared to rGluY, it was found that rGluY∆RICIN even at a concentration of 0.5 mg/mL (approximately 11.6 µM) could only inhibit 60% of the growth of C. albicans.…”

Section: Anti-fungal Activities Of Rgluy and Rgluy∆ricinmentioning

confidence: 99%

Novel Anti-Fungal d-Laminaripentaose-Releasing Endo-β-1,3-glucanase with a RICIN-like Domain from Cellulosimicrobium funkei HY-13

et al. 2021

View full text Add to dashboard Cite

Endo-β-1,3-glucanase plays an essential role in the deconstruction of β-1,3-d-glucan polysaccharides through hydrolysis. The gene (1650-bp) encoding a novel, bi-modular glycoside hydrolase family 64 (GH64) endo-β-1,3-glucanase (GluY) with a ricin-type β-trefoil lectin domain (RICIN)-like domain from Cellulosimicrobium funkei HY-13 was identified and biocatalytically characterized. The recombinant enzyme (rGluY: 57.5 kDa) displayed the highest degradation activity for laminarin at pH 4.5 and 40 °C, while the polysaccharide was maximally decomposed by its C-terminal truncated mutant enzyme (rGluYΔRICIN: 42.0 kDa) at pH 5.5 and 45 °C. The specific activity (26.0 U/mg) of rGluY for laminarin was 2.6-fold higher than that (9.8 U/mg) of rGluYΔRICIN for the same polysaccharide. Moreover, deleting the C-terminal RICIN domain in the intact enzyme caused a significant decrease (>60%) of its ability to degrade β-1,3-d-glucans such as pachyman and curdlan. Biocatalytic degradation of β-1,3-d-glucans by inverting rGluY yielded predominantly d-laminaripentaose. rGluY exhibited stronger growth inhibition against Candida albicans in a dose-dependent manner than rGluYΔRICIN. The degree of growth inhibition of C. albicans by rGluY (approximately 1.8 μM) was approximately 80% of the fungal growth. The superior anti-fungal activity of rGluY suggests that it can potentially be exploited as a supplementary agent in the food and pharmaceutical industries.

show abstract

Molecular cloning and anti-fungal effect of endo-β-1,3-glucanase from Thermotoga maritima

Cited by 11 publications

References 19 publications

Expression of the Thermobifida fusca β-1,3-Glucanase in Yarrowia lipolytica and Its Application in Hydrolysis of β-1,3-Glucan from Four Kinds of Polyporaceae

Expression of the Thermobifida fusca β-1,3-Glucanase in Yarrowia lipolytica and Its Application in Hydrolysis of β-1,3-Glucan from Four Kinds of Polyporaceae

Molecular Cloning and Biochemical Characterisation of a Novel Acidic Laminarinase Derived from Jermuk Hot Spring Metagenome

Novel Anti-Fungal d-Laminaripentaose-Releasing Endo-β-1,3-glucanase with a RICIN-like Domain from Cellulosimicrobium funkei HY-13

Contact Info

Product

Resources

About