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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biotechnology progress 9 (1993), S. 533-538 
    ISSN: 1520-6033
    Source: ACS Legacy Archives
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology reviews 16 (1995), S. 0 
    ISSN: 1574-6976
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract: The plant cell wall can be regarded as a giant bag-like macromolecule in which crystalline bundles of cellulose are embedded in a covalently linked matrix of hemicellulose and lignin. This heterologous polymer represents the dominant form of biomass on earth and a formidable challenge for solubilization and bioconversion. Bioconversion of lignocellulose requires the saccharification of both the hemicellulose and cellulose. Hemicellulose is composed of a mixture of sugars and can be readily hydrolysed by dilute acid at 140°C to produce a syrup containing pentoses and hexoses. However, no organisms in nature rapidly and efficiently convert both pentoses and hexoses into a single product of value. Our laboratory has developed such an organism by genetic engineering. Recombinant strains of Gram-negative bacteria (Escherichia coli or Klebsiella oxytoca or Erwinia sp.) have been constructed in which genes encoding the ethanol pathway from Zymomonas mobilis (pdc and adh) were inserted into the chromosome. These strains now efficiently convert all of the component sugars of hemicellulose and (cellulose) into ethanol. The saccharification of cellulose is more difficult and more complex. An enzymatic approach is preferred but at least three classes of enzymes are needed: endoglucanase, exoglucanase, and β-glucosidase. Klebsiella oxytoca and Erwinia sp. possess the native ability to transport and metabolize cellobiose (also cellotriose, xylobiose, and xylotriose), minimizing the need for added β-glucosidase. K. oxytoca strain P2, an ethanol-producing recombinant, has been evaluated in simultaneous saccharification and fermentation experiments to determine optimal conditions and limits of performance. Temperature was varied between 32 and 40°C over a pH range of 5.0–5.8 with 100 g 1−1 of crystalline cellulose (Sigmacell 50, Sigma Chemical Company, St. Louis, MO) as the substrate and commercial cellulase (Spezyme CE; Genencor, South San Francisco, CA). A broad optimum for fermentation was observed which allowed the production of over 44 g ethanol 1−1 (82–87% of the maximum theoretical yield). Two optimal saccharification and fermentation conditions were identified for fermentation yield, pH 5.2 at 35°C and pH 5.5 at 32°C, which produced 47 g ethanol 1−1 in 144 h (0.48 g ethanol (g cellulose) −1). Although yields were reduced at the lowest cellulase levels tested (2–5 filter paper units (g cellulose)−1), ethanol production per unit enzyme was much higher.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0006-3592
    Keywords: lignocellulose ; ethanol ; Klebisella oxytoca ; fermentation ; cellulase ; cellulose ; cellobiose ; biomass ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pretreatment of sugar cane bagasse is essential for a simultaneous saccharification and fermentation (SSF) process which uses recombinant Klebsiella oxytoca strain P2 and Genencor Spezyme CE. Strain P2 has been genetically engineered to express Zymomonas mobilis genes encoding the ethanol pathway and retains the native ability to transport and metabolize cellobiose (minimizing the need for extracellular cellobiase). In SSF studies with this organism, both the rate of ethanol production and ethanol yield were limited by saccharification at 10 and 20 filter papaer units (FPU) g-1 acid-treated bagasse. Dilute slurries of biomass were converted to ethanol more efficiently (over 72% of theoretical yield) in simple batch fermentations than slurries containing high solids albeit with the production of lower levels of ethanol. With high solids (i.e., 160 g acid-treated bagasse L-1), a combination of 20 FPU cellulase g-1 bagasse, preincubation under saccharification conditions, and additional grinding (to reduce particle size) were required to produce ca. 40 g ethanol L-1. Alternatively, almost 40 g ethanol L-1 was produced with 10 FPU cellulase g-1 bagasse by incorporating a second saccharification step (no further enzyme addition) followed by a second inoculation and short fermentation. In this way, a theoretical ethanol yield of over 70% was achieved with the production of 20 g ethanol 800 FPU-1 of commercial cellulase. © 1994 John Wiley & Sons, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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