The Experts below are selected from a list of 1986 Experts worldwide ranked by ideXlab platform
Montarop Yamabhai - One of the best experts on this subject based on the ideXlab platform.
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Mannan biotechnology: from biofuels to health
Critical Reviews in Biotechnology, 2015Co-Authors: Montarop Yamabhai, Suttipong Sak-ubol, Witsanu Srila, Dietmar HaltrichAbstract:Mannans of different structure and composition are renewable bioresources that can be widely found as components of lignocellulosic biomass in softwood and agricultural wastes, as non-starch reserve polysaccharides in endosperms and vacuoles of a wide variety of plants, as well as a major component of yeast cell walls. Enzymatic hydrolysis of Mannans using mannanases is essential in the pre-treatment step during the production of second-generation biofuels and for the production of potentially health-promoting manno-oligosaccharides (MOS). In addition, mannan-degrading enzymes can be employed in various biotechnological applications, such as cleansing and food industries. In this review, fundamental knowledge of mannan structures, sources and functions will be summarized. An update on various aspects of mannan-degrading enzymes as well as the current status of their production, and a critical analysis of the potential application of MOS in food and feed industries will be given. Finally, emerging areas of research on mannan biotechnology will be highlighted.
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efficient recombinant expression and secretion of a thermostable gh26 mannan endo 1 4 β mannosidase from bacillus licheniformis in escherichia coli
Microbial Cell Factories, 2010Co-Authors: Chomphunuch Songsiriritthigul, Dietmar Haltrich, Bancha Buranabanyat, Montarop YamabhaiAbstract:Mannans are one of the key polymers in hemicellulose, a major component of lignocellulose. The Mannan endo-1,4-β-mannosidase or 1,4-β-D-mannanase (EC 3.2.1.78), commonly named β-mannanase, is an enzyme that can catalyze random hydrolysis of β-1,4-mannosidic linkages in the main chain of Mannans, glucoMannans and galactoMannans. The enzyme has found a number of applications in different industries, including food, feed, pharmaceutical, pulp/paper industries, as well as gas well stimulation and pretreatment of lignocellulosic biomass for the production of second generation biofuel. Bacillus licheniformis is a Gram-positive endospore-forming microorganism that is generally non-pathogenic and has been used extensively for large-scale industrial production of various enzymes; however, there has been no previous report on the cloning and expression of mannan endo-1,4-β-mannosidase gene (manB) from B. licheniformis. The mannan endo-1,4-β-mannosidase gene (manB), commonly known as β-mannanase, from Bacillus licheniformis strain DSM13 was cloned and overexpressed in Escherichia coli. The enzyme can be harvested from the cell lysate, periplasmic extract, or culture supernatant when using the pFLAG expression system. A total activity of approximately 50,000 units could be obtained from 1-l shake flask cultures. The recombinant enzyme was 6 × His-tagged at its C-terminus, and could be purified by one-step immobilized metal affinity chromatography (IMAC) to apparent homogeneity. The specific activity of the purified enzyme when using locust bean gum as substrate was 1672 ± 96 units/mg. The optimal pH of the enzyme was between pH 6.0 - 7.0; whereas the optimal temperature was at 50 - 60°C. The recombinant β-mannanase was stable within pH 5 - 12 after incubation for 30 min at 50°C, and within pH 6 - 9 after incubation at 50°C for 24 h. The enzyme was stable at temperatures up to 50°C with a half-life time of activity (τ1/2) of approximately 80 h at 50°C and pH 6.0. Analysis of hydrolytic products by thin layer chromatography revealed that the main products from the bioconversion of locus bean gum and mannan were various manno-oligosaccharide products (M2 - M6) and mannose. Our study demonstrates an efficient expression and secretion system for the production of a relatively thermo- and alkali-stable recombinant β-mannanase from B. licheniformis strain DSM13, suitable for various biotechnological applications.
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cloning expression in pichia pastoris and characterization of a thermostable gh5 mannan endo 1 4 β mannosidase from aspergillus niger bk01
Microbial Cell Factories, 2009Co-Authors: Do Biencuong, Dietmar Haltrich, Dang Thithu, Jeanguy Berrin, To Kimanh, Jeanclaude Sigoillot, Montarop YamabhaiAbstract:Background Mannans are key components of lignocellulose present in the hemicellulosic fraction of plant primary cell walls. Mannan endo-1,4-β-mannosidases (1,4-β-D-mannanases) catalyze the random hydrolysis of β-1,4-mannosidic linkages in the main chain of β-Mannans. Biodegradation of β-Mannans by the action of thermostable mannan endo-1,4-β-mannosidase offers significant technical advantages in biotechnological industrial applications, i.e. delignification of kraft pulps or the pretreatment of lignocellulosic biomass rich in mannan for the production of second generation biofuels, as well as for applications in oil and gas well stimulation, extraction of vegetable oils and coffee beans, and the production of value-added products such as prebiotic manno-oligosaccharides (MOS).
Shigeo Suzuki - One of the best experts on this subject based on the ideXlab platform.
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distribution of antigenic oligomannosyl side chains in the cell wall Mannans of several strains of candida tropicalis
Archives of Microbiology, 2003Co-Authors: Hidemitsu Kobayashi, Hiroko Oyamada, Nobuyuki Shibata, Kyoko Matsuda, Shigeo SuzukiAbstract:In order to clarify the distribution of antigenic oligomannosyl side chains in the cell wall Mannans of the pathogenic yeast Candida tropicalis, the chemical structure of Mannans isolated from four C. tropicalis strains was investigated using nuclear magnetic resonance, two-dimensional homonuclear Hartmann-Hahn (2D-HOHAHA) spectroscopy. Two-dimensional maps of the 2D-HOHAHA clearly showed the distribution of oligomannosyl side chains in the Mannans. The linear side chain Manα1–3Manα1–(2Manα1–)n2Man [n≥2] is present in the Mannans from C. tropicalis IFO 0589 and IFO 1400, but not in the Mannans from IFO 0199 and IFO 1647. The mannan of IFO 0589 is the only mannan with the branched side chains, Manα1–3[Manα1–6]Manα1–(2Manα1-)n2Man and Manα1–2Manα1–3[Manα1–6]Manα1–(2Manα1-)n2Man [n≥2]. However, this mannan lacked the phosphate group and the β-1,2-linked oligomannosyl side chain which are features of this group. The Mannans of the C. tropicalis strains IFO 0589 and IFO 1400 possessed the side chains containing an α-1,3-linked mannose residue previously observed in Candida albicans.
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existence of novel branched side chains containing 1 2 and 1 6 linkages corresponding to antigenic factor 9 in the mannan of candida guilliermondii
Journal of Biological Chemistry, 1996Co-Authors: Nobuyuki Shibata, Yoshio Okawa, Hidemitsu Kobayashi, Akifumi Suzuki, Kyoko Ikuta, Kanehiko Hisamichi, Rieko Akagi, Tomoko Hosoya, Kumi Kawahara, Shigeo SuzukiAbstract:Abstract Isolation of β-linkage-containing side chain oligosaccharides from the mannan of Candida gilliermondii IFO 10279 strain has been conducted by acetolysis under mild conditions. A structural study of these oligosaccharides by one- and two-dimensional NMR and methylation analyses indicated the presence of extended oligosaccharide side chains with two consecutive β-1,2-linked mannose units at the nonreducing terminal of α-linked oligosaccharides. The linkage sequence present in this mannan, Manβ12Manα13Manα, has also been found in the mannan of Saccharomyces kluyveri but not in the mannan of Candida species. Furthermore, these oligosaccharides are branched at position 6 of the 3-O-substituted mannose units as follows. and The H-1 signals of the mannose units substituted by a 3,6-di-O-substituted unit showed a significant upfield shift (Δ = 0.04-0.08 ppm) due to a steric effect. The inhibition of an enzyme-linked immunosorbent assay between the mannan of C. guilliermondii and factor 9 serum with oligosaccharides obtained from several Mannans indicated that only the oligosaccharides with the above structure were active, suggesting that these correspond to the epitope of antigenic factor 9.
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existence of branched side chains in the cell wall mannan of pathogenic yeast candida albicans structure antigenicity relationship between the cell wall Mannans of candida albicans and candida parapsilosis
Journal of Biological Chemistry, 1995Co-Authors: Nobuyuki Shibata, Hidemitsu Kobayashi, Kyoko Ikuta, Tomonori Imai, Yohko Satoh, Richi Satoh, Atsuko Suzuki, Chizuko Kojima, Kanehiko Hisamichi, Shigeo SuzukiAbstract:Abstract Isolation of side chain oligosaccharides from Mannans of Candida albicans NIH B-792 (serotype B) and Candida parapsilosis IFO 1396 strains has been conducted by acetolysis under mild conditions. Structural study of these oligosaccharides by 1H and C NMR and methylation analyses indicated the presence of novel branched side chains with the following structures in C. albicans mannan.
Nobuyuki Shibata - One of the best experts on this subject based on the ideXlab platform.
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distribution of antigenic oligomannosyl side chains in the cell wall Mannans of several strains of candida tropicalis
Archives of Microbiology, 2003Co-Authors: Hidemitsu Kobayashi, Hiroko Oyamada, Nobuyuki Shibata, Kyoko Matsuda, Shigeo SuzukiAbstract:In order to clarify the distribution of antigenic oligomannosyl side chains in the cell wall Mannans of the pathogenic yeast Candida tropicalis, the chemical structure of Mannans isolated from four C. tropicalis strains was investigated using nuclear magnetic resonance, two-dimensional homonuclear Hartmann-Hahn (2D-HOHAHA) spectroscopy. Two-dimensional maps of the 2D-HOHAHA clearly showed the distribution of oligomannosyl side chains in the Mannans. The linear side chain Manα1–3Manα1–(2Manα1–)n2Man [n≥2] is present in the Mannans from C. tropicalis IFO 0589 and IFO 1400, but not in the Mannans from IFO 0199 and IFO 1647. The mannan of IFO 0589 is the only mannan with the branched side chains, Manα1–3[Manα1–6]Manα1–(2Manα1-)n2Man and Manα1–2Manα1–3[Manα1–6]Manα1–(2Manα1-)n2Man [n≥2]. However, this mannan lacked the phosphate group and the β-1,2-linked oligomannosyl side chain which are features of this group. The Mannans of the C. tropicalis strains IFO 0589 and IFO 1400 possessed the side chains containing an α-1,3-linked mannose residue previously observed in Candida albicans.
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existence of novel branched side chains containing 1 2 and 1 6 linkages corresponding to antigenic factor 9 in the mannan of candida guilliermondii
Journal of Biological Chemistry, 1996Co-Authors: Nobuyuki Shibata, Yoshio Okawa, Hidemitsu Kobayashi, Akifumi Suzuki, Kyoko Ikuta, Kanehiko Hisamichi, Rieko Akagi, Tomoko Hosoya, Kumi Kawahara, Shigeo SuzukiAbstract:Abstract Isolation of β-linkage-containing side chain oligosaccharides from the mannan of Candida gilliermondii IFO 10279 strain has been conducted by acetolysis under mild conditions. A structural study of these oligosaccharides by one- and two-dimensional NMR and methylation analyses indicated the presence of extended oligosaccharide side chains with two consecutive β-1,2-linked mannose units at the nonreducing terminal of α-linked oligosaccharides. The linkage sequence present in this mannan, Manβ12Manα13Manα, has also been found in the mannan of Saccharomyces kluyveri but not in the mannan of Candida species. Furthermore, these oligosaccharides are branched at position 6 of the 3-O-substituted mannose units as follows. and The H-1 signals of the mannose units substituted by a 3,6-di-O-substituted unit showed a significant upfield shift (Δ = 0.04-0.08 ppm) due to a steric effect. The inhibition of an enzyme-linked immunosorbent assay between the mannan of C. guilliermondii and factor 9 serum with oligosaccharides obtained from several Mannans indicated that only the oligosaccharides with the above structure were active, suggesting that these correspond to the epitope of antigenic factor 9.
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existence of branched side chains in the cell wall mannan of pathogenic yeast candida albicans structure antigenicity relationship between the cell wall Mannans of candida albicans and candida parapsilosis
Journal of Biological Chemistry, 1995Co-Authors: Nobuyuki Shibata, Hidemitsu Kobayashi, Kyoko Ikuta, Tomonori Imai, Yohko Satoh, Richi Satoh, Atsuko Suzuki, Chizuko Kojima, Kanehiko Hisamichi, Shigeo SuzukiAbstract:Abstract Isolation of side chain oligosaccharides from Mannans of Candida albicans NIH B-792 (serotype B) and Candida parapsilosis IFO 1396 strains has been conducted by acetolysis under mild conditions. Structural study of these oligosaccharides by 1H and C NMR and methylation analyses indicated the presence of novel branched side chains with the following structures in C. albicans mannan.
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structures of cell wall Mannans of pathogenic candida tropicalis ifo 0199 and ifo 1647 yeast strains
Infection and Immunity, 1994Co-Authors: H Kobayashi, Nobuyuki Shibata, Kyoko Matsuda, Atsuko Suzuki, M Suzuki, T Ikeda, Satoru Takahashi, S SuzukiAbstract:We conducted a structural analysis of the cell wall Mannans isolated from two Candida tropicalis strains, IFO 0199 and IFO 1647, exhibiting strong agglutinabilities against anti-Candida factor sera 5 and 6. The products released from these Mannans by acid treatment were identified as the oligosaccharides, from biose to pentaose, consisting solely of beta-1,2-linked mannopyranose units corresponding to common epitopes of Candida albicans serotypes A and B (factor 5). Mild acetolysis of acid- and alkali-treated Mannans produced large amounts of hexaose and heptaose, Man rho beta 1-2Man rho beta 1-2Man rho alpha 1-2Man rho alpha 1-2Man rho alpha 1-2Man and Man rho beta 1-2Man rho beta 1-2Man rho beta 1-2Man rho alpha 1-2 Man rho alpha 1-2Man, corresponding to the C. albicans serotype A-specific epitopes (factor 6). However, the homologous pentaose, Man rho beta 1-2Man rho alpha 1-2 Man, was not generated by this procedure. The oligosaccharides (biose to hexaose) obtained from the Mannans by conventional acetolysis were composed exclusively of alpha-1,2-linked mannopyranose units. Therefore, the Mannans of C. tropicalis IFO 0199 and IFO 1647 do not have the alpha-1,3-linked mannopyranose units previously observed in the Mannans of C. albicans and Candida stellatoidea. The results of this study and previous findings indicate that the similarity of the antigenicities of three Candida species, C. albicans serotype A, C. stellatoidea type II, and C. tropicalis, reside in the beta-1,2 and alpha-1,2 linkages containing oligomannosyl side chain (factor 6) in the cell wall mannan.
Phillip B. Pope - One of the best experts on this subject based on the ideXlab platform.
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human gut faecalibacterium prausnitzii deploy a highly efficient conserved system to cross feed on β mannan derived oligosaccharides
bioRxiv, 2020Co-Authors: Lars Jordhoy Lindstad, Leszek Michalak, Shaun Leivers, Gabriel V Pereira, Asmund K Rohr, Eric C Martens, Lauren S Mckee, Sylvia H Duncan, Bjorge Westereng, Phillip B. PopeAbstract:{beta}-Mannans are hemicelluloses that are abundant in modern diets as components in seed endosperms and common additives in processed food. Currently, the collective understanding of {beta}-mannan saccharification in the human colon is limited to a few keystone species, which presumably liberate low-molecular-weight mannooligosaccharide fragments that become directly available to the surrounding microbial community. Here we show that a dominant butyrate-producer in the human gut, Faecalibacterium prausnitzii, is able to acquire and degrade various {beta}-mannooligosaccharides ({beta}-MOS), which are derived by the primary mannanolytic activity of neighboring gut microbiota. Detailed biochemical analyses of selected protein components from their two {beta}-mannooligosaccharides ({beta}-MOS) utilization loci (FpMULs) supported a concerted model whereby the imported {beta}-MOS are stepwise disassembled intracellularly by highly adapted enzymes. Coculturing experiments of F. prausnitzii with the primary degrader Bacteroides ovatus on polymeric {beta}-mannan resulted in syntrophic growth and production of butyrate, thus confirming the high efficiency of the FpMULs uptake system. Genomic comparison with human F. prausnitzii strains and analyses of 2441 public human metagenomes revealed that FpMULs are highly conserved and distributed worldwide. Together, our results provide a significant advance in the knowledge of {beta}-Mannans metabolism and the degree to which its degradation is mediated by cross-feeding interactions between prominent beneficial microbes in the human gut. ImportanceCommensal butyrate-producing bacteria belonging to the Firmicutes phylum are abundant in the human gut and are crucial for maintaining health. Currently, insight is lacking into how they target otherwise indigestible dietary fibers and into the trophic interactions they establish with other glycan degraders in the competitive gut environment. By combining cultivation, genomic and detailed biochemical analyses this work reveals the mechanism enabling F. prausnitzii, as a model clostridial cluster IV Firmicute, to cross-feed and access {beta}-mannan-derived oligosaccharides released in the gut ecosystem by the action of primary degraders. A comprehensive survey of human gut metagenomes shows that FpMULs are ubiquitous in human populations globally, highlighting the importance of microbial metabolism of {beta}-Mannans/{beta}-MOS as a common dietary component. Our findings provide a mechanistic understanding of the {beta}-MOS utilization capability by F. prausnitzii that may be exploited to select dietary formulations specifically boosting this beneficial symbiont, thus butyrate production, in the gut.
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the human gut firmicute roseburia intestinalis is a primary degrader of dietary β Mannans
Nature Communications, 2019Co-Authors: Sabina Leanti La Rosa, Leszek Michalak, Maria Louise Leth, Morten Ejby Hansen, Nicholas A. Pudlo, Robert Glowacki, Gabriel Pereira, Christopher T. Workman, Magnus Ø. Arntzen, Phillip B. PopeAbstract:β-Mannans are plant cell wall polysaccharides that are commonly found in human diets. However, a mechanistic understanding into the key populations that degrade this glycan is absent, especially for the dominant Firmicutes phylum. Here, we show that the prominent butyrate-producing Firmicute Roseburia intestinalis expresses two loci conferring metabolism of β-Mannans. We combine multi-"omic" analyses and detailed biochemical studies to comprehensively characterize loci-encoded proteins that are involved in β-mannan capturing, importation, de-branching and degradation into monosaccharides. In mixed cultures, R. intestinalis shares the available β-mannan with Bacteroides ovatus, demonstrating that the apparatus allows coexistence in a competitive environment. In murine experiments, β-mannan selectively promotes beneficial gut bacteria, exemplified by increased R. intestinalis, and reduction of mucus-degraders. Our findings highlight that R. intestinalis is a primary degrader of this dietary fiber and that this metabolic capacity could be exploited to selectively promote key members of the healthy microbiota using β-mannan-based therapeutic interventions.
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The human gut Firmicute Roseburia intestinalis is a primary degrader of dietary β-Mannans
Nature Publishing Group, 2019Co-Authors: Sabina Leanti La Rosa, Leszek Michalak, Maria Louise Leth, Morten Ejby Hansen, Nicholas A. Pudlo, Robert Glowacki, Gabriel Pereira, Christopher T. Workman, Magnus Ø. Arntzen, Phillip B. PopeAbstract:How dietary β-Mannans are utilized by gut Gram-positive bacteria is unclear. Here, the authors uncover the enzymatic pathway for β-mannan metabolism in Roseburia intestinalis and show that these polysaccharides promote beneficial gut bacteria, highlighting a potential for β-mannan-based therapeutic interventions
Dietmar Haltrich - One of the best experts on this subject based on the ideXlab platform.
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Mannan biotechnology: from biofuels to health
Critical Reviews in Biotechnology, 2015Co-Authors: Montarop Yamabhai, Suttipong Sak-ubol, Witsanu Srila, Dietmar HaltrichAbstract:Mannans of different structure and composition are renewable bioresources that can be widely found as components of lignocellulosic biomass in softwood and agricultural wastes, as non-starch reserve polysaccharides in endosperms and vacuoles of a wide variety of plants, as well as a major component of yeast cell walls. Enzymatic hydrolysis of Mannans using mannanases is essential in the pre-treatment step during the production of second-generation biofuels and for the production of potentially health-promoting manno-oligosaccharides (MOS). In addition, mannan-degrading enzymes can be employed in various biotechnological applications, such as cleansing and food industries. In this review, fundamental knowledge of mannan structures, sources and functions will be summarized. An update on various aspects of mannan-degrading enzymes as well as the current status of their production, and a critical analysis of the potential application of MOS in food and feed industries will be given. Finally, emerging areas of research on mannan biotechnology will be highlighted.
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efficient recombinant expression and secretion of a thermostable gh26 mannan endo 1 4 β mannosidase from bacillus licheniformis in escherichia coli
Microbial Cell Factories, 2010Co-Authors: Chomphunuch Songsiriritthigul, Dietmar Haltrich, Bancha Buranabanyat, Montarop YamabhaiAbstract:Mannans are one of the key polymers in hemicellulose, a major component of lignocellulose. The Mannan endo-1,4-β-mannosidase or 1,4-β-D-mannanase (EC 3.2.1.78), commonly named β-mannanase, is an enzyme that can catalyze random hydrolysis of β-1,4-mannosidic linkages in the main chain of Mannans, glucoMannans and galactoMannans. The enzyme has found a number of applications in different industries, including food, feed, pharmaceutical, pulp/paper industries, as well as gas well stimulation and pretreatment of lignocellulosic biomass for the production of second generation biofuel. Bacillus licheniformis is a Gram-positive endospore-forming microorganism that is generally non-pathogenic and has been used extensively for large-scale industrial production of various enzymes; however, there has been no previous report on the cloning and expression of mannan endo-1,4-β-mannosidase gene (manB) from B. licheniformis. The mannan endo-1,4-β-mannosidase gene (manB), commonly known as β-mannanase, from Bacillus licheniformis strain DSM13 was cloned and overexpressed in Escherichia coli. The enzyme can be harvested from the cell lysate, periplasmic extract, or culture supernatant when using the pFLAG expression system. A total activity of approximately 50,000 units could be obtained from 1-l shake flask cultures. The recombinant enzyme was 6 × His-tagged at its C-terminus, and could be purified by one-step immobilized metal affinity chromatography (IMAC) to apparent homogeneity. The specific activity of the purified enzyme when using locust bean gum as substrate was 1672 ± 96 units/mg. The optimal pH of the enzyme was between pH 6.0 - 7.0; whereas the optimal temperature was at 50 - 60°C. The recombinant β-mannanase was stable within pH 5 - 12 after incubation for 30 min at 50°C, and within pH 6 - 9 after incubation at 50°C for 24 h. The enzyme was stable at temperatures up to 50°C with a half-life time of activity (τ1/2) of approximately 80 h at 50°C and pH 6.0. Analysis of hydrolytic products by thin layer chromatography revealed that the main products from the bioconversion of locus bean gum and mannan were various manno-oligosaccharide products (M2 - M6) and mannose. Our study demonstrates an efficient expression and secretion system for the production of a relatively thermo- and alkali-stable recombinant β-mannanase from B. licheniformis strain DSM13, suitable for various biotechnological applications.
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cloning expression in pichia pastoris and characterization of a thermostable gh5 mannan endo 1 4 β mannosidase from aspergillus niger bk01
Microbial Cell Factories, 2009Co-Authors: Do Biencuong, Dietmar Haltrich, Dang Thithu, Jeanguy Berrin, To Kimanh, Jeanclaude Sigoillot, Montarop YamabhaiAbstract:Background Mannans are key components of lignocellulose present in the hemicellulosic fraction of plant primary cell walls. Mannan endo-1,4-β-mannosidases (1,4-β-D-mannanases) catalyze the random hydrolysis of β-1,4-mannosidic linkages in the main chain of β-Mannans. Biodegradation of β-Mannans by the action of thermostable mannan endo-1,4-β-mannosidase offers significant technical advantages in biotechnological industrial applications, i.e. delignification of kraft pulps or the pretreatment of lignocellulosic biomass rich in mannan for the production of second generation biofuels, as well as for applications in oil and gas well stimulation, extraction of vegetable oils and coffee beans, and the production of value-added products such as prebiotic manno-oligosaccharides (MOS).
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Hydrolysis of isolated coffee mannan and coffee extract by mannanases of Sclerotium rolfsii.
Journal of biotechnology, 2000Co-Authors: Alois Sachslehner, Gabriele Foidl, Nikolaus Foidl, Georg Gübitz, Dietmar HaltrichAbstract:Different mannanase preparations obtained from the filamentous fungus Sclerotium rolfsii were used for the hydrolysis of coffee mannan, thus reducing significantly the viscosity of coffee extracts. Mannan is the main polysaccharide component of these extracts and is responsible for their high viscosity, which negatively affects the technological processing of instant coffee. Coffee mannan was isolated from green defatted Arabica beans by delignification, acid wash and subsequent alkali extraction with a yield of 12.8%. Additionally, coffee extract polysaccharides were separated by alcohol precipitation and were found to form nearly half of the coffee extract dry weight. These isolated Mannans as well as the mannan in the coffee extract were efficiently hydrolysed by the S. rolfsii mannanase, which resulted in significant viscosity reductions. Concurrently, the reducing sugar content increased continuously due to the release of various mannooligosaccharides including mannotetraose, mannotriose, and mannobiose. Both a partially purified, immobilised and a soluble, crude mannanase preparation were successfully employed for the degradation of coffee mannan.