Exploring Levansucrase Operon Regulating Levan-Type Fructooligosaccharides (L-FOSs) Production in HL12.

Levan biopolymer and levan-type fructooligosaccharides (L-FOSs) are β-2,6-linked fructans that have been used as non-digestible dietary fiber and prebiotic oligosaccharides in food and cosmeceutical applications. In this study, we explore the operon responsible for levan and L-FOSs production in HL12. Presented is the first genomic perspective on sucrose utilization and the levan biosynthesis pathway in this bacterium.

Characterization of cold-active trehalose synthase from Pseudarthrobacter sp. for trehalose bioproduction.

Trehalose is a functional sugar that has numerous applications in food, cosmetic, and pharmaceutical products. Production of trehalose from maltose via a single-step enzymatic catalysis using trehalose synthase (TreS) is a promising method compared with the conventional two-step process due to its simplicity with lower formation of byproducts. In this study, a cold-active trehalose synthase (PaTreS) from Pseudarthrobacter sp.

Enhancement of catalytic activity and alkaline stability of cellobiohydrolase by structure-based protein engineering.

Unlabelled: Alkaline cellobiohydrolases have the potential for application in various industries, including pulp processing and laundry where operation under high pH conditions is preferred. In this study, variants of Cel6A cellobiohydrolase from were generated by structural-based protein engineering with the rationale of increasing catalytic activity and alkaline stability. The variants included removal of the carbohydrate-binding module (CBM) and substitution of residues 173 and 200.

Functional Characterization of Recombinant Endo-Levanase (LevBk) from Bacillus koreensis HL12 on Short-Chain Levan-Type Fructooligosaccharides Production.

Levan-type fructooligosaccharides (L-FOSs) are a prominent class of non-digestible oligosaccharides with potential as nutritional prebiotics. Endo-levanase, which randomly hydrolyzes β-(2,6)-linkages in fructans, is a promising enzyme for short-chain FOS production. In this work, a recombinant levanase (LevBk) from Bacillus koreensis strain HL12 was characterized.

Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in .

Concerns over climate change have led to increased interest in renewable fuels in recent years. Microbial production of advanced fuels from renewable and readily available carbon sources has emerged as an attractive alternative to the traditional production of transportation fuels. Here, we engineered the yeast , an industrial powerhouse in heterologous enzyme production, to produce the advanced biofuel isobutanol from sugarcane trash hydrolysates.

A novel low temperature active maltooligosaccharides-forming amylase from HL12 as biocatalyst for maltooligosaccharide production.

Unlabelled: Maltooligosaccharide-forming amylases (MFAses) are promising enzymes for a variety of industrial applications. In this study, a maltooligosaccharide-forming amylase (BkAmy) isolated from HL12 was first heterologous expressed and characterized. According to structural-sequence alignment, BkAmy contained seven conserved regions which are the signature of a novel GH13 subfamily.

Profiling multi-enzyme activities of strains growing on various agro-industrial residues.

Unlabelled: Agro-industrial wastes provide potential sources of carbon for production of fungal enzymes applied for various biotechnological applications. In this study, 23 strains of were systematically investigated for their capability on production of carbohydrate-processing enzymes used in industries. The strains were grown on glucose or selected agricultural wastes comprising varied chemical compositions as the sole carbon source.

Biochemical characterization of xylanase GH11 isolated from Aspergillus niger BCC14405 (XylB) and its application in xylooligosaccharide production.

Objective: To develop an endo-β-1,4-xylanase with high specificity for production of prebiotic xylooligosaccharides that optimally works at moderate temperature desirable to reduce the energy cost in the production process.

Results: The xylB gene, encoding for a glycosyl hydrolase family 11 xylanase from a thermoresistant fungus, Aspergillus niger BCC14405 was expressed in a methylotrophic yeast P. pastoris KM71 in a secreted form.

Screening, Cloning, Expression and Characterization of New Alkaline Trehalose Synthase from and Its Application for Trehalose Production.

Trehalose is a non-reducing disaccharide in increasing demand for applications in food, nutraceutical, and pharmaceutical industries. Single-step trehalose production by trehalose synthase (TreS) using maltose as a starting material is a promising alternative process for industrial application due to its simplicity and cost advantage. TBRC 1196 was identified using the developed screening method as a potent strain for TreS production.

Identification and characterization of a novel AA9-type lytic polysaccharide monooxygenase from a bagasse metagenome.

Lytic polysaccharide monooxygenases (LPMOs) are auxiliary enzymes catalyzing oxidative cleavages of cellulose chains in crystalline regions, resulting in their increasing accessibility to the hydrolytic enzyme counterparts and hence higher released sugars from biomass saccharification. In this study, a novel auxiliary protein family 9 LPMO (BgAA9) was identified from a metagenomic library derived from a thermophilic microbial community in bagasse collection site where diverse AA9 and AA10 putative sequences were annotated. The enzyme showed highest similarity to a glycoside hydrolase family 61 from Chaetomium thermophilum.

Designing cellulolytic enzyme systems for biorefinery: From nature to application.

Cellulolytic enzymes play a key role on conversion of lignocellulosic plant biomass to biofuels and biochemicals in sugar platform biorefineries. In this review, we survey composite carbohydrate-active enzymes (CAZymes) among groups of cellulolytic fungi and bacteria that exist under aerobic and anaerobic conditions. Recent advances in designing effective cellulase mixtures are described, starting from the most complex microbial consortium-based enzyme preparations, to single-origin enzymes derived from intensively studied cellulase producers such as Trichoderma reesei, Talaromyces cellulolyticus, and Penicellium funiculosum, and the simplest minimal enzyme systems comprising selected sets of mono-component enzymes tailor-made for specific lignocellulosic substrates.

Development of tailor-made synergistic cellulolytic enzyme system for saccharification of steam exploded sugarcane bagasse.

Designing a tailor-made synergistic system is a promising strategy for developing an effective enzyme for saccharification of lignocellulosic materials. In this study, a cellulolytic enzyme mixture comprising selected core recombinant enzymes for hydrolysis of sugarcane bagasse pretreated by alkaline-catalyzed steam explosion was optimized using a mixture design approach. The optimized enzyme system comprised a cellobiohydrolase (Cel7A) from Talaromyces cellulolyticus, an endo-glucanase (Cel7B) from Thielavia terrestris, a β-glucosidase (BGL) and an endo-β1,4-xylanase (XYN) from Aspergillus aculeatus at the ratio of 0.

Synergistic action of recombinant accessory hemicellulolytic and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation.

Synergism between core cellulases and accessory hydrolytic/non-hydrolytic enzymes is the basis of efficient hydrolysis of lignocelluloses. In this study, the synergistic action of three recombinant accessory enzymes, namely GH62 α-l-arabinofuranosidase (ARA), CE8 pectin esterase (PET), and GH10 endo-1,4-beta-xylanase (XYL) from Aspergillus aculeatus expressed in Pichia pastoris to a commercial Trichoderma reesei cellulase (Accellerase® 1500; ACR) on hydrolysis of alkaline pretreated rice straw was studied using a mixture design approach. Applying the full cubic model, the optimal ratio of quaternary enzyme mixture was predicted to be ACR:ARA:PET:XYL of 0.

Binding characteristics and synergistic effects of bacterial expansins on cellulosic and hemicellulosic substrates.

Expansins are non-catalytic proteins which loosen plant cell wall structure. In this study, binding kinetics and synergistic action of five bacterial expansins on cellulosic and hemicellulosic polysaccharides were studied. The expansins differed in binding capacity (Bmax) and affinity (Kd) for different substrates.

Identification of novel bacterial expansins and their synergistic actions on cellulose degradation.

Novel expansins, non-catalytic proteins which induce weakening of the rigid cellulose structure, have been identified in this study. A pipeline of bioinformatics was implemented for sequence and structure-based prediction of putative bacterial expansin-like group × family from NR databases. All putative expansins had no detectable activity against cellulosic and hemicellulosic substrates but showed varying degrees of synergy (2.

Optimisation of synergistic biomass-degrading enzyme systems for efficient rice straw hydrolysis using an experimental mixture design.

Synergistic enzyme system for the hydrolysis of alkali-pretreated rice straw was optimised based on the synergy of crude fungal enzyme extracts with a commercial cellulase (Celluclast™). Among 13 enzyme extracts, the enzyme preparation from Aspergillus aculeatus BCC 199 exhibited the highest level of synergy with Celluclast™. This synergy was based on the complementary cellulolytic and hemicellulolytic activities of the BCC 199 enzyme extract.

Insights into the phylogeny and metabolic potential of a primary tropical peat swamp forest microbial community by metagenomic analysis.

A primary tropical peat swamp forest is a unique ecosystem characterized by long-term accumulation of plant biomass under high humidity and acidic water-logged conditions, and is regarded as an important terrestrial carbon sink in the biosphere. In this study, the microbial community in the surface peat layer in Pru Toh Daeng, a primary tropical peat swamp forest, was studied for its phylogenetic diversity and metabolic potential using direct shotgun pyrosequencing of environmental DNA, together with analysis of 16S rRNA gene library and key metabolic genes. The community was dominated by aerobic microbes together with a significant number of facultative and anaerobic microbial taxa.

Identification and characterization of lipolytic enzymes from a peat-swamp forest soil metagenome.

In this work, a metagenomic library was generated from peat-swamp forest soil obtained from Narathiwat Province, Thailand. From a fosmid library of approximately 15,000 clones, six independent clones were found to possess lipolytic activity at acidic pH. Analysis of pyrosequencing data revealed six ORFs, which exhibited 34-71% protein similarity to known lipases/esterases.