Amsonia spp. as potential fuel crops for arid lands

The biomass of three desert plants, Amsonia kearneyana, A. grandiflora and A. palmeri, was used for the production of glucose and ethanol by simultaneous saccharification and fermentation techniques. Ethanol yields were 0.46 g g^-1 for A. keurneyana, 0.51 g g^-1 for A. grandiflora and 0.51 g g^-1 for A. palmeri. When the plant materials were saccharified into glucose only, the yields obtained were 0.35 g g^-1 for A. kearneyana, 0.39 g g^-1 for A. grandiflora and 0.22 g g^-1 for A. palmeri.H. Punnapayak is with the Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand; J.J. Hoffmann is with the Bioresources Research Facility, University of Arizona, Tuscon, AZ 85706, USA.     

Energy Metabolism in Rhizomes of Acorus calamus (L.) and in Tubers of Solanum tuberosum (L.) With Regard to their Anoxia Tolerance

Rhizomes of the marsh plant Acorus calamus (L.) and tubers of the flooding-intolerant Solanum tuberosum (L.) var. Bintje, both kept under strict anoxia, differ markedly in their fermentation properties. The fermentation capacities as measured by ADH and LDH activities and their respective product concentrations were estimated. While rhizomes of Acorus calamus, having high ADH and low LDH activities, accumulate mainly ethanol, tubers of Solanum tuberosum tend towards lactic acid fermentation. The total amount of adenine nucleotides is quite stable in Acorus calamus, whereas they show a sharp decline in S. tuberosum during the first 6h of anoxia. The adenylate energy charge of A. calamus recovers after a short initial drop (AEC > 0.8). AEC values of S. tuberosum decrease rapidly and remain at very low values (AEC ～ 0.3). Tuber tissues became soft and lost viability after about 48–72 h of anoxia at 25 °C. This might be due to tissue acidification and impaired energy metabolism, but not to the lack of energy reserves. Energy metabolism of A. calamus is well adapted to anoxia.     

Development and evaluation of Trichoderma asperellum preparation for control of sheath blight of rice (Oryza sativa L.)

Sheath blight, which is caused by Rhizoctonia solani, is a disease that majorly impacts rice production. A biocontrol agent used for control rice sheath blight must be sprayed on the stem at specific times during rice growth, a process that is labour-intensive and renders the antagonist vulnerable to environmental factors. In this study, Trichoderma asperellum T12 was used to produce preparation by solid-state fermentation using a surface-response method. Rice hull was selected as a carrier based on its ability to sustain the T12 floating in the water and protect T12 from ultraviolet irradiation. The production of a T12-based preparation required 32% wheat bran, 7% inoculum, 2.3 g kg^?1 (NH₄)₂SO₄ and 65% water content, with fermentation at 27.5°C for 30 days and agitation every six days. The preparation demonstrated 90% biocontrol efficacy and significantly (P > 0.05) increased the seed-set rate and 1000-grain weight as compared with the pathogen treatment. The population of Trichoderma on the surface of rice leaf sheath in the treatment applied with T12 preparation increased from 232 cfu (colony forming units) g^?1 fw (fresh weight) to 436 cfu g^?1 fw during rice growth stage, which was significantly (P > 0.05) higher than pathogen treatment. The population of R. solani on the leaf sheath increased from 41 cfu g^?1 fw to 271 cfu g^?1 fw in the pathogen treatment, while remained stable (P > 0.05) at level of 10–23 cfu g^?1 fw in T12 preparation applied treatment. Biocontrol of sheath blight by the addition of the preparation to the soil is effective and decreases the costs of agro-industrial waste disposal.     

Frequency of the Insertion Sequence IS4Bsu1 among Bacillus subtilis Strains Isolated from Fermented Soybean Foods in Southeast Asia

Among 45 Bacillus subtilis strains isolated from non-salted types of fermented soybeans produced in several Southeast Asian countries, 20 had the insertion sequence IS4Bsu1 in the chromosome. In contrast, none of 49 B. subtilis strains of non-food origin contained IS4Bsu1. Frequent occurrence of this mobile DNA element in the soybean-fermenting B. subtilis would reflect the fact that few strains flourish on soybeans and thereby contribute to soybean fermentation.     

Flooding Induced Increase in Alcohol Dehydrogenase Activity in Timothy and Ryegrass Seedlings

Two forage grasses, timothy (Phleum pratense L.) and ryegrass (Lolium multiflorum Lam.) were exposed to flooding, and activities of alcohol dehydrogenase (ADH) and their isozyme profiles were determined. The flooding stress increased ADH activities in both species. This increase was 2-times greater in timothy than in ryegrass. Only one ADH isozyme was found in non-flooded seedlings of both species, whereas two and four bands were identified in ryegrass and timothy seedlings, respectively, under flooding stress. This revised version was published online in June 2006 with corrections to the Cover Date.     

Application of nuclear magnetic resonance spectroscopy to analysis of ethanol fermentation kinetics in yeasts and bacteria

Nuclear Magnetic Resonance (NMR) spectroscopy is proving to be a very valuable technique for characterizing the metabolic status of a range of microbial fermentations. This non-invasive method allows us not only to determine the presence of particular metabolites, but also to monitor reaction rates, enzyme activities and transport mechanisms in vivo. Despite the low levels of the carbon-13 isotope (1.1%), natural-abundance ¹³C-NMR studies have proven useful in monitoring the progress of various fermentation processes. Furthermore, ³¹P-NMR can provide noninvasive information relating to cellular metabolism, and on the energy status of the cells. This results from the facility with NMR to identify various nucleotide phosphates and other energy-rich compounds in the cell, as well as to characterize changes in the intracellular pH from the chemical shifts of internal phosphate and other phosphorylated intermediates. In this review, we will summarize the use of NMR as an analytical tool in biotechnology and also discuss examples that illustrate how NMR can be used to obtain significant information on the characteristics of ethanol fermentations in both yeasts and bacteria.     

A chemically-defined medium for production of compactin by Penicillium citrinum

A mutant strain of Penicillium citrinum grown in a chemically-defined production medium, yielded 145 mg compactin l^–1. The medium also facilitated spectrophotometric analysis of compactin. Addition of KH₂PO₄in the production medium did not increase the compactin production, while addition of a surfactant, Tween 80, increased compactin to 175 mg l^–1. Inoculation with 10⁷ spores ml^–1 and initial pH of 6.5–7 were the most suitable for compactin production.     

Aerobic submerged fermentation by acetic acid bacteria for vinegar production: Process and biotechnological aspects

Strictly aerobic acetic acid bacteria (AAB) have a long history of use in fermentation processes, and the conversion of ethanol to acetic acid for the production of vinegar is the most well-known application.At the industrial scale, vinegar is mainly produced by submerged fermentation, which refers to an aerobic process in which the ethanol in beverages such as spirits, wine or cider is oxidized to acetic acid by AAB. Submerged fermentation requires robust AAB strains that are able to oxidize ethanol under selective conditions to produce high-titer acetic acid. Currently submerged fermentation is conducted by unselected AAB cultures, which are derived from previous acetification stocks and maintained by repeated cultivation cycles.In this work, submerged fermentation for vinegar production is discussed with regard to advances in process optimization and parameters (oxygen availability, acetic acid content and temperature) that influence AAB activity. Furthermore, the potential impact arising from the use of selected AAB is described.Overcoming the acetification constraints is a main goal in order to facilitate innovation in submerged fermentation and to create new industry-challenging perspectives.     

Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum

Declining fossil fuel reserves, coupled with environmental concerns over their continued extraction and exploitation have led to strenuous efforts to identify renewable routes to energy and fuels. One attractive option is to convert glycerol, a by-product of the biodiesel industry, into n-butanol, an industrially important chemical and potential liquid transportation fuel, using Clostridium pasteurianum. Under certain growth conditions this Clostridium species has been shown to predominantly produce n-butanol, together with ethanol and 1,3-propanediol, when grown on glycerol. Further increases in the yields of n-butanol produced by C. pasteurianum could be accomplished through rational metabolic engineering of the strain. Accordingly, in the current report we have developed and exemplified a robust tool kit for the metabolic engineering of C. pasteurianum and used the system to make the first reported in-frame deletion mutants of pivotal genes involved in solvent production, namely hydA (hydrogenase), rex (Redox response regulator) and dhaBCE (glycerol dehydratase). We were, for the first time in C. pasteurianum, able to eliminate 1,3-propanediol synthesis and demonstrate its production was essential for growth on glycerol as a carbon source. Inactivation of both rex and hydA resulted in increased n-butanol titres, representing the first steps towards improving the utilisation of C. pasteurianum as a chassis for the industrial production of this important chemical.     

Cellulose depolymerization to glucose and other water soluble polysaccharides by shear deformation and high pressure treatment

The simultaneous action of shear deformation and high pressure (SDHP) creates changes in the structure of wood and its main components (cellulose, hemicelluloses, lignin). The formation of water and alkali soluble polysaccharides under SDHP action, proceeds in seconds in the solid state, without the use of any reagents and solvents. Therefore, SDHP seems to be a technologically safe method and friendly to the environment. The amorphization of cellulose crystallites and depolymerization of cellulose chains were observed under a wide range of pressures (1–6 GPa), both for cellulose samples and the cellulose part of wood. Similar depolymerization occurs in the hemicellulose part of wood. The decomposition of polysaccharides under SDHP causes the formation of the water soluble part, whose content increases with pressure and the applied shear deformation. A maximum solubility of 40% and 55% was registered at 6 GPa following treatment of cellulose and birch wood samples. A higher output in the case of wood can be explained by a specific role of lignin under SDHP, which acts as a ‘grinding stone’ during cellulose and hemicelluloses destruction. As shown by high-performance size exclusion chromatography, the water soluble fraction obtained from cellulose contained glucose (2.6%), cellobiose (9.6%), cellotriose (16.6%) and other higher water soluble oligomers (71%). Almost complete dissolution (98%) of the treated cellulose sample can be achieved by extraction with 10% NaOH solution. The SDHP treated birch wood was subjected to submerged fermentation (with Trichoderma viride), and a 13% output of proteins was obtained. In this case, the water soluble part played the role of the so called ‘start sugars’. Abbreviations: ASF, alkali soluble fraction; DP, degree of polymerization; EC, energy consumption; HP, high pressure; LMWS, low molecular weight sugars; MC, moisture content; MCC, microcrystalline cellulose; SD, shear deformation, SDHP, shear deformation under high pressure; SS, shear strength; WSF, water soluble fraction This revised version was published online in November 2006 with corrections to the Cover Date.