Abstract Effectively releasing the locked polysaccharides from recalcitrant lignocellulose to fermentable sugars is among greatest technical and economic barriers realization of biorefineries because leading pre‐treatment technologies suffer low sugar yields, and/or severe reaction conditions, high cellulase use, narrow substrate applicability, capital investment, etc. A new featuring modest conditions (50°C atmospheric pressure) was demonstrated fractionate amorphous cellulose,...

ADVERTISEMENT RETURN TO ISSUEPREVNoteA Transition from Cellulose Swelling to Dissolution by o-Phosphoric Acid: Evidence Enzymatic Hydrolysis and Supramolecular StructureY.-H. Percival Zhang, Jingbiao Cui, Lee R. Lynd, Lana KuangView Author Information Biological Systems Engineering Department, Virginia Polytechnic Institute State University, Blacksburg, 24060, Center for Nanomaterials Research, Dartmouth College, Hanover, New Hampshire 03755, Thayer School of Engineering, 03755 Cite this:...

A new functionally based kinetic model for enzymatic hydrolysis of pure cellulose by the Trichoderma cellulase system is presented. The represents actions cellobiohydrolases I, cellobiohydrolase II, and endoglucanase I; incorporates two measurable physically interpretable substrate parameters: degree polymerization (DP) fraction β-glucosidic bonds accessible to cellulase, Fa (Zhang Lynd, 2004). Initial enzyme-limited reaction rates simulated are consistent with several important behaviors...

The bioenergetics of cellulose utilization by Clostridium thermocellum was investigated. Cell yield and maintenance parameters, \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}Y_{X/{\mathrm{ATP}}}^{{\mathrm{True}}}=16.44\end{equation*}\end{document} g cell/mol ATP m = 3.27 mmol ATP/g cell per hour, were obtained from...

Specific cellulose hydrolysis rates (g of cellulose/g cellulase per h) were shown to be substantially higher (2.7- 4.7-fold) for growing cultures Clostridium thermocellum as compared with purified preparations from this organism in controlled experiments involving both batch and continuous cultures. This “enzyme–microbe synergy” requires the presence metabolically active cellulolytic microbes, is not explained by removal products bulk fermentation broth, appears due surface phenomena...

Abstract Cell‐free synthetic (enzymatic) pathway biotransformation (SyPaB) is the assembly of a number purified enzymes (usually more than 10) and coenzymes for production desired products through complicated biochemical reaction networks that single enzyme cannot do. SyPaB, as compared to microbial fermentation, has several distinctive advantages, such high product yield, great engineering flexibility, titer, fast rate. Biocommodities (e.g., ethanol, hydrogen, butanol) are low‐value where...

Cocktail reception: Biohydrogen is produced in high yield from cellulosic materials and water a one-pot process catalyzed by up to 14 enzymes one coenzyme. This assembly of results non-natural catabolic pathways. These spontaneous reactions are conducted under modest reaction conditions (32 degrees C atmospheric pressure).

Cost-efficient production of sustainably-derived biochemicals and biofuels is a critical goal modern biotechnology. The current predominant biotransformation method mainly based on microbial fermentation, but this system suffers from mismatch engineering cellular objectives, appropriates significant fraction substrate/energy sources for self-replication, poses scaling challenges. Cell-free biosystems biomanufacturing (CFB2)—complex, cell-free systems that catalyze the conversion renewable...

We developed a general restriction enzyme-free and ligase-free method for subcloning up to three DNA fragments into any location of plasmid. The multimer generated by prolonged overlap extension PCR was directly transformed in Escherichia coli [e.g., TOP10, DH5α, JM109, BL21(DE3)] Bacillus subtilis obtaining chimeric plasmids.

Summary Bacillus subtilis can serve as a powerful platform for directed evolution, especially secretory enzymes. However, cloning and transformation of DNA mutant library in B. are not easy they Escherichia coli . For direct , here we developed new protocol based on supercompetent cells prepared from the recombinant strain SCK6 multimeric plasmids. This is simple (restriction enzyme‐, phosphatase‐ ligase‐free), fast (i.e. 1 day) high efficiency ∼10 7 or 4 transformants per µg plasmid ligated...

Myo-Inositol (vitamin B8) is widely used in the drug, cosmetic, and food & feed industries. Here, we present an vitro non-fermentative enzymatic pathway that converts starch to inositol one vessel. This comprised of four enzymes operate without ATP or NAD+ supplementation. All enzyme BioBricks are carefully selected from hyperthermophilic microorganisms, is, alpha-glucan phosphorylase Thermotoga maritima, phosphoglucomutase Thermococcus kodakarensis, 1-phosphate synthase Archaeoglobus...

Abstract The switchgrass (SG) samples pretreated by cellulose solvent‐ and organic solvent‐based lignocellulose fractionation were characterized enzymatic hydrolysis, substrate accessibility assay, scanning electron microscopy, X‐ray diffraction (XRD), cross polarization/magic angle spinning (CP/MAS) 13 C nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR). Glucan digestibility of the SG was 89% at hour 36 one filter paper unit cellulase per gram glucan....

One-step purification of a multi-enzyme complex was developed based on mixture cell extracts containing three dockerin-containing enzymes and one family 3 cellulose-binding module (CBM3)-containing scaffoldin through high-affinity adsorption low-cost solid regenerated amorphous cellulose (RAC). The three-enzyme complex, called synthetic metabolon, self-assembled the interaction between dockerin in each enzyme cohesins scaffoldin. metabolons were either immobilized external surface RAC or...

Growing populations and climate change pose great challenges to food security. Humankind is confronting a serious question: how will we feed the world in near future? This study presents an out-of-the-box solution involving highly efficient biosynthesis of artificial starch microbial proteins from available abundant agricultural residue as new sources. A one-pot biotransformation using vitro coenzyme-free synthetic enzymatic pathway baker's yeast can simultaneously convert dilute sulfuric...

Abstract Enzymatic electrosynthesis has gained more and interest as an emerging green synthesis platform, particularly for the fixation of CO 2 . However, simultaneous utilization a nitrogenous molecule enzymatic value‐added products never been reported. In this study, we constructed in vitro multienzymatic cascade based on reductive glycine pathway demonstrated electrocatalytic system that allowed conversion NH 3 sole carbon nitrogen sources to synthesize glycine. Through effective coupling...

Biofilm formation enhances bacterial survival and antibiotic tolerance, but the underlying mechanisms are incompletely understood. Here, we show that biofilm growth is accompanied by a reduction in energy metabolism membrane potential, together with metabolic exchanges between inner outer regions biofilms. More specifically, nutrient-starved cells interior supply amino acids to periphery, while peripheral experience decrease potential provide fatty cells. Fatty facilitate repair of...

Efficient liberation of fermentable soluble sugars from lignocellulosic biomass waste not only decreases solid handling but also produces value-added biofuels and biobased products. Industrial hemp, a special economic crop, is cultivated for its high-quality fibers high-value seed oil, hollow stalk cords (hurds) are cellulosic waste. The cellulose-solvent-based lignocellulose fractionation (CSLF) technology has been developed to separate components under modest reaction conditions (Zhang,...

Production of biofuels and biobased products from lignocellulosic materials is an emerging industry. Quantitative saccharification (QS) a widely used method for determining carbohydrate composition in materials. The NREL methods (versions 1996 2006) involve primary hydrolysis 72% w/w sulfuric acid at 30 °C, converting polysaccharides into oligosaccharides, secondary 4% 121 all oligosaccharides to monomeric sugars. Because some are degraded during the harsh processes, sugar control set sugars...

Effective hydrolysis of pretreated lignocellulose mediated by cellulase requires an in-depth understanding adsorption and desorption. Here we developed a simple method for determining the adsorbed on cellulosic materials or lignocellulose, which involves (i) in presence 10 M NaOH at 121 °C 20 min, (ii) ninhydrin assay amino acids released from hydrolyzed cellulase. The major lignocellulosic components (i.e., cellulose, hemicellulose, lignin) did not interfere with assay. A number desorption...