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Twenty-Seventh Symposium on Biotechnology for Fuels and Chemicals

James D. McMillan ; William S. Adney ; Jonathan R. Mielenz ; K. Thomas Klasson (eds.)

Resumen/Descripción – provisto por la editorial

No disponible.

Palabras clave – provistas por la editorial

Biotechnology

Disponibilidad
Institución detectada Año de publicación Navegá Descargá Solicitá
No detectada 2006 SpringerLink

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Tipo de recurso:

libros

ISBN impreso

978-1-58829-866-9

ISBN electrónico

978-1-59745-268-7

Editor responsable

Springer Nature

País de edición

Reino Unido

Fecha de publicación

Información sobre derechos de publicación

© Humana Press Inc. 2006

Cobertura temática

Tabla de contenidos

Enzymatic Synthesis of Sorbitan Methacrylate According to Acyl Donors

Gwi-Taek Jeong; Hye-Jin Lee; Hae-Sung Kim; Don-Hee Park

Recently, sugar polymers have been considered for use as biomaterials in medical applications. These biomaterials are already used extensively in burn dressings, artificial membranes, and contact lenses. In this study, we investigated the optimum conditions under which the enzymatic synthesis of sorbitan methacrylate can be affected using Novozym 435 in t -butanol from sorbitan and several acyl donors (ethyl methacrylate, methyl methacrylate, and vinyl methacrylate). The enzymatic synthesis of sorbitan methacrylate, catalyzed by Novozym 435 in t -butanol, reached an approx 68% conversion yield at 50 g/L of 1,4-sorbitan, 5% (w/v) of enzyme content, and a 1:5 molar ratio of sorbitan to ethyl methacrylate, with a reaction time of 36 h. Using methyl methacrylate as the acyl donor, we achieved a conversion yield of approx 78% at 50 g/L of 1,4-sorbitan, 7% (w/v) of enzyme content, at a 1:5 molar ratio, with a reaction time of 36 h. Sorbitan methacrylate synthesis using vinyl methacrylate as the acyl donor was expected to result in a superior conversion yield at 3% (w/v) of enzyme content, and at a molar ratio greater than 1:2.5. Higher molar ratios of acyl donor resulted in more rapid conversion rates. Vinyl methacrylate can be applied to obtain higher yields than are realized when using eathyl methacrylate or methyl methacrylate as acyl donors in esterification reactions catalyzed by Novozym 435 in organic solvents. Enzyme recycling resulted in a drastic reduction in conversion yields.

Palabras clave: Immobilized enzyme; bioconversion; optimization; biocatalysis; sorbitan; esterification.

Session 1B - Enzyme Catalysis and Engineering | Pp. 265-277

Effect of Inhibitors Released During Steam-Explosion Pretreatment of Barley Straw on Enzymatic Hydrolysis

MA Prado García-Aparicio; Ignacio Ballesteros; Alberto González; José Miguel Oliva; Mercedes Ballesteros; MA José Negro

The influence of the liquid fraction (prehydrolysate) generated during steam-explosion pretreatment (210°C, 15 min) of barley straw on the enzymatic hydrolysis was determined. Prehydrolysate was analyzed for degradation compounds and sugars’ content and used as a medium for enzymatic hydrolysis tests after pH adjusting to 4.8. Our results show that the presence of the compounds contained in the prehydrolysate strongly affects the hydrolysis step (a 25% decrease in cellulose conversion compared with control). Sugars are shown to be more potent inhibitors of enzymatic hydrolysis than degradation products.

Palabras clave: Enzymatic hydrolysis; barley straw; inhibition; steam explosion.

Session 1B - Enzyme Catalysis and Engineering | Pp. 278-288

Purification and Characterization of Two Xylanases From Alkalophilic and Thermophilic Bacillus licheniformis 77-2

Valquiria B. Damiano; Richard Ward; Eleni Gomes; Heloiza Ferreira Alves-Prado; Roberto Da Silva

The alkalophilic bacteria Bacillus licheniformis 77-2 produces significant quantities of thermostable cellulase-free xylanases. The crude xylanase was purified to apparent homogeneity by gel filtration (G-75) and ionic exchange chromatography (carboxymethyl sephadex, Q sepharose, and Mono Q), resulting in the isolation of two xylanases. The molecular masses of the enzymes were estimated to be 17 kDa (X-I) and 40 kDa (X-II), as determined by SDS-PAGE. The K _m and V _max values were 1.8 mg/mL and 7.05 U/mg protein (X-I), and 1.05 mg/mL and 9.1 U/mg protein (X-II). The xylanases demonstrated optimum activity at pH 7.0 and 8.0–10.0 for xylanase X-I and X-II, respectively, and, retained more than 75% of hydrolytic activity up to pH 11.0. The purified enzymes were most active at 70 and 75°C for X-I and X-II, respectively, and, retained more than 90% of hydrolytic activity after 1 h of heating at 50°C and 60°C for X-I and X-II, respectively. The predominant products of xylan hydrolysates indicated that these enzymes were endoxylanases.

Palabras clave: Xylanase; Bacillus licheniformis; xylanase purification; alkalophilic bacteria; xylanase characterization.

Session 1B - Enzyme Catalysis and Engineering | Pp. 289-302

Oxidation Capacity of Laccases and Peroxidases as Reflected in Experiments With Methoxy-Substituted Benzyl Alcohols

Feng Hong; Leif J. Jönsson; Knut Lundquist; Yijun Wei

A set of methoxy-substituted benzyl alcohol (MBA) congeners were examined regarding susceptibility to oxidation by Trametes versicolor laccase, T. versicolor lignin peroxidase and horseradish peroxidase: 2,4,5-trimethoxybenzyl alcohol (TMBA), 3,4,5-TMBA, 2,3,4-TMBA, 2,5-dimethoxybenzyl alcohol (DMBA), 3,4-DMBA, and 2,3-DMBA. The corresponding methoxy-substituted benzaldehydes were strongly predominant as products on enzymic oxidation. This together with different reaction rates and redox potentials makes the MBAs suitable as substrates in the characterization of ligninolytic enzymes. For fungal laccase, the reaction rate order was: 2,4,5-TMBA ≫ 2,5-DMBA > 3,4-DMBA > 3,4,5-TMBA ∼ 2,3,4-TMBA ∼ 2,3-DMBA. Horseradish peroxidase displayed a similar reactivity order. Oxidation of some of the MBAs with laccase and horseradish peroxidase was only observed when the reactions were carried out at low pH and with relatively high-substrate concentration. 3,4-DMBA (veratryl alcohol) was the best substrate for lignin peroxidase and the reaction rate order was: 3,4-DMBA > 2,4,5-TMBA ∼ 3,4,5-TMBA > 2,5-DMBA > 2,3,4-TMBA ∼ 2,3-DMBA. The oxidation experiments with different MBAs elucidate the potential of the enzymes as oxidants in various applications.

Palabras clave: Lignin peroxidase; horseradish peroxidase; laccase; methoxy-substituted benzyl alcohols.

Session 1B - Enzyme Catalysis and Engineering | Pp. 303-319

Obtainment of Chelating Agents Through the Enzymatic Oxidation of Lignins by Phenol Oxidase

Gabriela M. M. Calabria; Adilson R. Gonçalves

Oxidation of lignin obtained from acetosolv and ethanol/water pulping of sugarcane bagasse was performed by phenol oxidases: tyrosinase (TYR) and laccase (LAC), to increase the number of carbonyl and hydroxyl groups in lignin, and to improve its chelating capacity. The chelating properties of the original and oxidized lignins were compared by monitoring the amount of Cu^2+ bound to lignin by gel permeation chromatography. The Acetosolv lignin oxidized with TYR was 16.8% and with LAC 21% higher than that of the original lignin. For ethanol/water lignin oxidized with TYR was 17.2% and with LAC 18% higher than that of the original lignin.

Palabras clave: Lignin; enzymatic oxidation; polyphenoloxidase; chelating agents.

Session 1B - Enzyme Catalysis and Engineering | Pp. 320-325

Reuse of the Xylanase Enzyme in the Biobleaching Process of the Sugarcane Bagasse Acetosolv Pulp

Luís R. M. Oliveira; Regina Y. Moriya; Adilson R. Gonçalves

In this work, pretreatment-enzymatic series of the bagasse-sugarcane pulp and alkaline extraction of enzyme treated pulp were carried out. In the pretreatment an enzyme dose was utilized and acetosolv pulp suspension of 3% (w/v) with different solvents (distilled water, 0.05 mol/L acetate buffer pH 5.5 and 0.05 mol/L phosphate buffer pH 7.25) stirred at 85 rpm for 2 or 4 h. The enzymes used were pulpzyme and cartazyme, both commercial. The accompaniment of the enzymatic activity was carried out through measurement in initial and finish of each enzymatic pretreatment. The xylanase-treated pulps and xylanase-alkaline-extracted pulps were analyzed regarding kappa number and viscosity. Pulpzyme recovery was better in phosphate buffered medium (84, 46, and 23% for first, second, and third enzymatic treatment, respectively) although in aqueous medium reached only 2% for every treatments. However, the improvement of pulp properties was evidenced only in aqueous medium for pulpzyme. Cartazyme recovery was similar for both solvents (water and acetate buffer), reaching values around 19% for first enzymatic treatment and 9% for second one. Nevertheless, the pulp properties increased only in acetate buffered medium.

Palabras clave: Enzymatic pretreatment; xylanase; acetosolv pulping; sugarcane bagasse.

Session 1B - Enzyme Catalysis and Engineering | Pp. 326-333

Detection of Nisin Expression by Lactococcus lactis Using Two Susceptible Bacteria to Associate the Effects of Nisin With EDTA

Thereza Christina Vessoni Penna; Angela Faustino Jozala; Thomas Rodolfo Gentille; Adalberto Pessoa; Olivia Cholewa

Nisin, a bacteriocin produced during the exponential growth phase of Lactococcus lactis ATCC 11454, inhibits the growth of a broad range of Grampositive bacteria. Gram-negative bacteria can also be inhibited by nisin with EDTA. In this study, nisin production was assayed by the agar diffusion method using Lactobacillus sake ATCC 15521 and a recombinant Escherichia coli DH5-α expressing the recombinant green fluorescent protein as the nisin-susceptible test organisms. The titers of nisin expressed and released in culture media were quantified and expressed in arbitrary units (AU/mL of medium) and converted to standard nisin concentration (Nisaplin®, 25 mg of pure nisin with an activity of 1 × 10^6 AU/mL). The expression and release of nisin by L. lactis in skimmed milk (9.09% total solids) with Man Rugosa Shepeer-Bacto Lactobacilli broth (1:1) was monitored in a 5 L New Brunswick fermentor. Combining EDTA with nisin increased the bactericidal effect of nisin on the bacteria examined. The presence of EDTA was necessary to inhibit E. coli growth with nisin. L. sake was shown to be a good indicator for the evaluation of nisin release in the culture media, including with the addition of EDTA.

Palabras clave: Nisin; Lactococcus lactis; Lactobacillus sake; EDTA; Escherichia coli; recombinant green fluorescent protein.

Session 1B - Enzyme Catalysis and Engineering | Pp. 334-346

Introduction to Session 2

Paris Tsobanakis

Session 2 focused on the transition from the biorefineries of today to the ones of the future. The various presenters provided an international perspective, offering several visions about how future biorefineries may look, what technologies they will require, and what products they will produce. One recurring high-level theme was the need to logically capitalize on the learning being accrued in our existing grain, sugar, pulp, and other plant biomass processing mills, which represent today’s biorefineries. Such facilities are now being used to produce a large number of products, including starches, sugars, oils, fuels, organic acids, polymers, and power. Ethanol and biodiesel production, in particular, are experiencing tremendous growth and are benefiting from increased “critical mass” and significant economies of scale. Biorefineries based on ethanol and biodiesel production provide routes for dramatically increasing the use of renewable resources to supply fuels and chemicals for the global economy.

Palabras clave: Apply Biochemistry; Global Economy; Renewable Resource; Critical Mass; Submerged Cultivation.

Session 2 - Today’s Biorefineries | Pp. 347-348

Existing Biorefinery Operations That Benefit From Fractal-Based Process Intensification

Vadim Kochergin; Mike Kearney

Ion exchange, adsorption, and chromatography are examples of separation processes frequently used in today’s biorefineries. The particular tasks for which these technologies can be successfully applied are highly influenced by capital cost and efficiency. There exists a potential for significantly increasing the efficiency of these processes whereas simultaneously decreasing their size and capital cost. This potential for process intensification can be realized with the use of engineered fractal equipment. The cost savings potential and the possibilities for broadening the use of fractal-based separation technologies in future biorefinery concepts is illustrated by examples of full-scale implementation in the sugar and sweetener industries.

Palabras clave: Biorefinery; fractal; liquid separations; chromatography; ion exchange.

Session 2 - Today’s Biorefineries | Pp. 349-360

The Importance of Utility Systems in Today’s Biorefineries and a Vision for Tomorrow

Tim Eggeman; Dan Verser

Heat and power systems commonly found in today’s corn processing facilities, sugar mills, and pulp and paper mills will be reviewed. We will also examine concepts for biorefineries of the future. We will show that energy ratio, defined as the ratio of renewable energy produced divided by the fossil energy input, can vary widely from near unity to values greater than 12. Renewable-based utility systems combined with low-fossil input agricultural systems lead to high-energy ratios.

Palabras clave: Ethanol; indirect ethanol process; green power.

Session 2 - Today’s Biorefineries | Pp. 361-381