Evaluation of structure and hydrolysis activity of Candida rugosa Lip7 in presence of sub-/super-critical CO2

This work aimed to assess the effect of sub-/super-critical CO₂ on the structure and activity of Candida rugosa Lip7 (CRL7) in its solution form. The structure was examined by SDS-PAGE gel electrophoresis, circular dichroism (CD) and fluorescence spectra photometry. Results revealed that the primary structure remained intact after sub-/super-critical CO₂ treatment, and the secondary structure altered at the pressure of 10 MPa and temperature 40 °C for 30 min incubation, but it was reflex to its native form with increasing incubation time up to 150 min under 10 MPa and 40 °C. Meanwhile, the tertiary structure via fluorescence spectra analysis showed that the intensity of the maximal emission wavelength at 338 nm decreased under the conditions of 10 MPa and 40 °C for 150 min. Furthermore, the residue hydrolysis activity and kinetics constants (V_max and K_m) of CRL7 treated with sub-/super-critical CO₂ were also investigated. In cases of 6 MPa and 35 °C, or 10 MPa and 40 °C for 30 min, activity variance of CRL7 was maybe caused by its secondary structure alteration. But in case of 10 MPa and 40 °C for 150 min, the tertiary structure change was perhaps responsibility for CRL7 activity enhancement.     

Behavioral and mortality responses of the bivalves Scrobicularia plana and Cerastoderma edule to temperature,as indicator of climate change's potential impacts

Temperature is one of the most important abiotic factors affected by climate change. It determines physiological processes, ecological patterns and establishes the limits of geographic distribution of species. The induced thermal stress frequently results in physiological and behavioral responses and, in extreme cases, may lead to mortality episodes. Scrobicularia plana and Cerastoderma edule behavioral and mortality responses to temperature were evaluated. Specimens were sampled in the Mondego estuary (Portugal), acclimated and exposed to different temperature treatments (5–35 °C). Individual activity and mortality were registered during 120 h laboratory assays. Both species showed a thermal optimum for their activity (S. plana: 15–23 °C; C. edule: 20–23 °C), and survival was mainly affected by high temperature (S. plana: LC50_120 h = 28.86 °C; C. edule: LC50_120 h = 28.01 °C), with 100% mortality above critical values (≥32 °C). Results further indicated that both species are more affected the higher the temperature and the longer the exposure time. This study indicates that the occurrence of extreme climatic events, especially heat waves, may be particularly impairing for these species.     

Stability and activity of Dictyoglomus thermophilum GH11 xylanase and its disulphide mutant at high pressure and temperature

The functional properties of extremophilic Dictyoglomus thermophilum xylanase (XYNB) and the N-terminal disulphide-bridge mutant (XYNB-DS) were studied at high pressure and temperature. The enzymes were quite stable even at the pressure of 500 MPa at 80 °C. The half-life of inactivation in these conditions was over 30 h. The inactivation at 80 °C in atmospheric pressure was only 3-times slower. The increase of pressure up to 500 MPa at 80 °C decreased only slightly the enzyme's stability, whereas in 500 MPa the increase of temperature from 22 to 80 °C decreased significantly more the enzyme's stability. While the high temperature (80–100 °C) decreased the enzyme reaction with short xylooligosaccharides (xylotetraose and xylotriose), the high pressure (100–300 MPa) had an opposite effect. The temperature of 100 °C strongly increased the K_m but did not affect the k_cat to the same extent, thus indicating that the interaction of the substrate with the active site suffers before the catalytic reaction begins to decrease as the temperature rises. Circular dichroism spectroscopy showed the high structural stability of XYNB and XYNB-DS at 93 °C.     

Engineering a high-performance,metagenomic-derived novel xylanase with improved soluble protein yield and thermostability

The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340 U/mg) and broad pH active range of 5.5–10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7-TS) showed a 250-fold increase in half-life at 55 °C, with a 10 °C increase in optimal temperature compared to that of wild-type XYL7. A truncated enzyme (XYL7-Tr3) acquired by protein engineering showed similar catalytic properties as the wild-type, with a tenfold increase in soluble protein yield by the mutant. The reducing sugar produced by XYL7-TC was about fourfold greater than that produced by their parents when incubated with xylan at 60 °C for 4 h. The engineered novel xylanase exhibited superior enzymatic performance and showed promise as an excellent candidate for industrial application due to its high specific activity, stability and soluble protein yield.     

Biocontrol of major postharvest pathogens on apple using Rhodotorula glutinis and its effects on postharvest quality parameters

The biocontrol activity of Rhodotorula glutinis on gray mold decay and blue mold decay of apple caused by Botrytis cinerea and Penicillium expansum, respectively, was investigated, as well as its effects on postharvest quality of apple fruits. The results show there was a significant negative correlation between concentrations of the yeast cells and the disease incidence of the pathogens. The higher concentration of the R. glutinis, the better effect of the biocontrol capacity. At concentrations of R. glutinis 1 × 10⁸ CFU ml^?1, the amount of gray mold decay was completely inhibited after 5 days incubation at 20 °C, after challenge with B. cinerea spores suspension of 1 × 10⁵ spores ml^?1; While the blue mold decay was completely inhibited at concentrations of 5 × 10⁸ CFU ml^?1, at challenged with P. expansum spores suspension of 5 × 10⁴ spores ml^?1_. These results demonstrated that the efficacy of R. glutinis in controlling of gray mold decay of apples was better than the efficacy of controlling blue mold. R. glutinis within inoculated wounds on apples increased in numbers at 20 °C from an initial level of 9.5 × 10⁵ CFU per wound to 2.24 × 10⁷ CFU at 20 °C after 1 day. The highest population of the yeast was recovered 4 days after inoculation, the yeast population in wounds increased by 56.9 times. After that, the population of the yeast began to decline very slowly. R. glutinis significantly reduced the incidence of natural infections on intact fruit from 75% in the control fruit to 28.3% after 5 days at 20 °C, and from 58.3 to 6.7% after 30 days at 4 °C followed by 4 days at 20 °C. R. glutinis treatment had no deleterious effect on quality parameters after 5 days at 20 °C or after 30 days at 4 °C followed by 4 days at 20 °C.     

Ecophysiological requirements on seed germination of a Mediterranean perennial grass (Stipa tenacissima L.) under controlled temperatures and water stress

Stipa tenacissima L. (alpha grass) steppes are one of the most representative ecosystems in arid Mediterranean ecosystems. On the one hand these steppes, which are perpetually exposed to climate and strong anthropogenic pressure, have undergone severe degradation. On the other hand, the ability of S. tenacissima to regenerate naturally is significantly reduced. In this study the germination response and seedling emergence of S. tenacissima are examined in relation to the main environmental factors (water stress and temperature) under laboratory-controlled conditions. The main aim of this paper was to investigate the influence of temperature over a temperature range (10 °C–30 °C) and water stress induced by the solutions of polyethylene glycol (PEG)-6000 (0 to − 1.6 MPa) for a period of 30 days, on the germination behavior of S. tenacissima seeds. The results showed that temperatures between 10° and 20 °C seem to be favorable for the germination of this species, with optimum temperatures among accessions found in 20 °C. When seeds were water-stressed, germination severely decreased at − 0.8 MPa, indicating that the accession resistance limits to the water stress, and was completely inhibited at − 1.6 MPa. Consequently, the final germination percentage (FGP) decreased and the mean time germination (MTG) increased. Based on the empirical data of the germination rate, we estimated that the parameters of the thermal time and hydrotime models showed different values in all accessions which proves the difference between accession adaptive capacities.     

High hydrostatic pressure increased stability and activity of immobilized lipase in hexane

Lipases are important to high value product synthesis, modification, and enhancement. However, they are often unstable above 40 °C. While most current applications of high hydrostatic pressure (HHP) are for inactivating deleterious enzymes, there is evidence that HHP can stabilize and increase activity of some enzymes. This study examines the apparent kinetics of immobilized lipase-catalyzed synthesis of isoamyl acetate at HHP in hexane. HHP reduced thermal inactivation of lipase by up to 152% after 4 h at 80 °C and 400 MPa when compared to incubations at low pressure. No significant differences were found in activation energy (E_a) at different pressures, irrespectively of the pressurization and heating sequence, and were between 35.7 ± 3.5 and 47.8 ± 8.2 kJ mol^?1, depending on the method. In all methods utilized, activity at 63.5 and 80 °C at 400 MPa was greater (from about 20 to 96% increase) than at low pressure. Activity increased by 110% at low pressure versus a 239% increase at 350 MPa when the temperature was increased from 40 to 80 °C. Increasing pressure up to 350 MPa increased lipase activity while pressures greater than 350 MPa maintained or decreased lipase activity. Activation volume (ΔV^≠) appeared negative between ambient pressure and 200 MPa in contrast to a positive ΔV^≠ between 300 and 600 MPa. Apparent ΔV^≠ was 14.3 ± 1.7 or 15.2 ± 2.2 cm³ mol^?1 at 40 or 80 °C, respectively, between 300 and 500 MPa.     

Cold storage of the egg parasitoids Trissolcus basalis (Wollaston) and Telenomus podisi Ashmead (Hymenoptera: Scelionidae)

Adults of Trissolcus basalis and Telenomus podisi were stored either at 15 or 18 °C after their immature development had been completed at 18 or 25 °C. Longevity of the parasitoids in the storage temperatures was evaluated, as well as fecundity and longevity following their return to 25 °C after different periods in reproductive diapause. Temperature during immature development influenced female longevity and highest mean longevity was obtained for females that developed to the adult stage at 25 °C and then were stored at 15 °C (ca. 13 months for T. basalis and 10 months for Te. podisi). For adults of T. basalis that developed at 25 °C, storage periods of 120 or 180 days at 15 or 18 °C did not affect fecundity. The fecundity of T. basalis females that developed at 18 °C and were stored for 120 days at 15 or 18 °C was not affected; however, after remaining for 180 days, fecundity was reduced in ca. 30 and 50%, respectively. Storage of Te. podisi adults at 15 or 18 °C significantly reduced fecundity. It is concluded that adults of T. basalis can be stored in the adult stage at 15 or 18 °C between two soybean crop seasons for mass production purposes, aiming the biological control of stink bugs.     

Effects of constant temperature,exposure period,and age on diapause induction in Trichogramma dendrolimi

Trichogramma dendrolimi can be successfully reproduced in fresh eggs dissected from ovaries of the Chinese tussah silkworm (Antheraea pernyi) and is widely used in biological control of lepidopteran agricultural and forest pests in China. Diapause induction of T. dendrolimi in A. pernyi eggs was investigated through exposing the parasitoid to six constant temperatures (16, 13, 10, 7, 4, and 1 °C) for 19 exposure periods between 10 and 46 days. The sensitive age of T. dendrolimi for diapause induction was explored through a separate experiment to examine the parasitoids that had developed for 2, 3, 4, 5, 6, 7, and 8 days at 26 °C after parasitization, under the six constant temperatures, respectively. Diapause was induced at 10 or 7 °C, and the induction period was 4–6 weeks. The sensitive age of T. dendrolimi to react at the induction temperature was 2–3 days (at 26 °C). At 7 and 10 °C, the diapause rate increased with increasing exposure period and decreased with increased T. dendrolimi age at exposure. The optimum method to induce diapause in T. dendrolimi consisted of exposing hosts for parasitization at 26 °C for 8 h, and then keeping them at 26 °C for 40 h, finally, moving them into 10 °C for 4 weeks.     

Temperature-dependent development of Lista haraldusalis (Walker) (Lepidoptera: Pyralidae) on Platycarya strobilacea

The effect of constant temperatures on development and survival of Lista haraldusalis (Walker) (Lepidoptera: Pyralidae), a newly reported insect species used to produce insect tea in Guizhou province (China), was studied in laboratory conditions at seven temperatures (19 °C, 22 °C, 25 °C, 28 °C, 31 °C, 34 °C, and 37 °C) on Platycarya strobilacea. Increasing the temperature from 19 °C to 31 °C led to a significant decrease in the developmental time from egg to adult emergence, and then the total developmental time increased at 34 °C. Egg incubation was the stage where L. haraldusalis experienced the highest mortality at all temperatures. The survival of L. haraldusalis was significantly higher at 25 °C and 28 °C, whereas none of the eggs hatched at 37 °C. Common and Ikemoto linear models were used to describe the relationship between the temperature and the developmental rate for each immature stage of L. haraldusalis. The estimated values of the lower temperature threshold and thermal constant of the total immature stages using Common and Ikemoto linear models were 11.34 °C and 11.20 °C, and 939.85 and 950.41 degree-days, respectively. Seven nonlinear models were used to fit the experimental data to estimate the developmental rate of L. haraldusalis. Based on the biological significance for model evaluation, Ikemoto linear, Logan-6, and SSI were the best models that fitted each immature stage of L. haraldusalis and they were used to estimate the temperature thresholds. These thermal requirements and temperature thresholds are crucial for facilitating the development of factory-based mass rearing of L. haraldusalis.     

Thermal resistance time of estuarine fishes Etroplus suratensis and Therapon jarbua

The resistance time of Etroplus suratensis (Bloch) and Therapon jarbua (Forsskal) were determined for fish acclimated to a series of temperatures 20°C, 25°C, 28°C, 30°C, and 35°C. The results indicated that these two species of fishes showed increased thermal resistance when acclimation temperatures were increased or decreased, respectively. The biokinetic range of temperature for E. suratensis is from 12°C to 46°C and that for T. jarbua is from 8°C to 44°C.     

Smoke-water effect on the germination of Amazonian tree species

Smoke stimulates seed germination of a range of species from ecosystems that may or may not be fire prone. We evaluated the effects of smoke-water on germination of ten tree species of economic value in the Amazon region. Two materials were burnt to produce smoke-water: germination paper and the wood of Cecropia palmata Willd. Seven dilutions of the solutions were tested. Seeds of nine forest trees were germinated under controlled laboratory conditions (25 °C ± 2 °C) in the laboratory. Bertholletia excelsa Humb. & Bonpl., was tested in the nursery (approximately 25–36 °C) because of its large seeds. Irrespective of the material burned, smoke-water significantly increased seed germination of three species: Cordia goeldiana Hub., Ochroma pyramidale (Cav. ex Lam.) Urb. and Jacaranda copaia (Aubl.) D. Don. and there was a significant inhibitory effect on Swietenia macrophylla King. Germination was accelerated by smoke in J. copaia, B. excelsa and Bellucia grossularioides (L.) Triana. The most pronounced effect was observed in B. excelsa, as the mean germination time of 108 d (control) was reduced to 76 d with smoke-water made from germination paper (dilution of 1:25) and to 61 d with the one from Cecropia wood (dilution of 1:250). For five of the ten species studied, smoke-water either increased or accelerated seed germination, irrespective of the materials used for its production. Seeds with low vigour and prolonged germination time seemed to be more receptive to smoke.     

Effect of preharvest spraying Cryptococcus laurentii on postharvest decay and quality of strawberry

Application of a suspension (log 8.0/ml) of Cryptococcus laurentii prior to harvest led to a reduction in Botrytis cinerea decay of strawberries stored at 4 or 20 °C, for 12 or 4 days, respectively. The frequency of spraying antagonist significantly influenced disease incidence in strawberry fruit. The best inhibition of disease was achieved when fruit sprayed C. laurentii with three applications that began 6 days prior to harvest, and the incidence of gray mold and natural decay treated with this method was 21% and 11%, compared with 88% and 62% in the control after storage at 20 °C for 4 days. A similar result occurred in the treated fruit after storage at 4 °C for 12 days. Dilation plate counts on Rose Bengal agar and scanning electron microscopy results showed that three applications with C. laurentii at 6, 3 and 0 days before harvest improved its ability to colonize the epidermis of strawberry fruit in the greenhouse and during storage compared to single application. In addition, antagonistic yeast spraying before harvest could reduce the weight loss, delay the decrease of the firmness and ascorbic acid, but had no significant effect on the contents of soluble solids, titratable acidity and fruit surface color. These results suggested that preharvest spraying with C. laurentii might be a promising alternative to fungicide application for decay control of strawberry.     

Effects of different temperatures on the life history of Evania appendigaster L. (Hymenoptera: Evaniidae), a solitary oothecal parasitoid of Periplaneta americana L. (Dictyoptera: Blattidae)

The influence of temperatures on the life parameters of the solitary oothecal parasitoid Evania appendigaster, was investigated in the laboratory. Parasitized oothecae of Periplaneta americana were left to develop under seven constant temperatures: 15, 17, 20, 25, 30, 35, and 40 °C. At the end, we found that: (i) E. appendigaster was able to complete development within the temperature range of 17–34 °C; (ii) mean adult longevity decreased as temperature increased, with the temperature of 40 °C being fatal in a matter of hours; (iii) males lived longer than females between 15 and 30 °C; (iv) adult emergence rate was the highest at 25 °C, and (v) no wasps emerged at 15 or 40 °C. Non-emerged oothecae contained either unhatched eggs or dead larvae. We determined the theoretical lower developmental threshold and thermal constant for the complete development as 12.9 °C and 584.8 day-degrees for males, and 13.1 °C and 588.2 day-degrees for females, respectively. A good balance between faster development, maximum adult longevity and good egg viability was obtained between 25–30 °C, and that would be the best temperature range for rearing E. appendigaster.     

Tetracycline-resistant bacteria as indicators of antimicrobial resistance in protected waters—The example of the Drwęca River Nature Reserve (Poland)

The objective of this study was to determine the suitability of Tet^R tetracycline-resistant bacteria as potential indicators of drug resistance, a parameter of the microbiological quality of river waters in natural reserves which are threatened by man-made pollution. The microbiological assays covered a 15-km long section of the upper reach of the Drw?ca River (Poland), a part of the European Ecological “Natura 2000” Network of nature protected areas. The quality of the investigated ecosystem was affected by surface runoffs from the river's agricultural catchment as well as outflows from three fish farms. The counts of Tet^R bacteria, incubated at 14 °C and 28 °C on TSA medium with sheep blood and tetracycline, were determined in river water samples. The highest counts of both bacterial groups were determined in samples collected from sites behind fish farms. A statistical analysis of the abundance of Tet^R14 °C and Tet^R28 °C bacteria revealed significant differences in the size of Tet^R28 °C populations at the studied sampling sites (p = 0.0011), which is why hemolytic bacteria of this group (HemTet^R28 °C) were selected for further investigations. The predominant strains in the group of 86 HemTet^R28 °C isolates obtained by 16S rRNA gene sequencing were Pseudomonas fluorescens (42 isolates) and Aeromonas hydrophila (28 isolates). Analyses of the identified HemTet^R28 °C strains demonstrated MIC ≥256 μg/ml in more than 50% isolates. The MAR index of HemTet^R28 °C was in the range of 0.67 at the control site to 1 at sites behind fish farms. The results suggest that tetracycline-resistant bacteria, in particular HemTet^R28 °C, are a reliable indicator of antimicrobial resistance and the microbial quality of surface waters polluted due to human activity. The above can be attributed to several factors: (I) the highest percentage share of HemTet^R28 °C among HPC28 °C was noted at sites behind fish farms, (II) tetracycline-resistant bacteria quickly respond to environmental changes, as demonstrated by the high level of resistance to tetracycline and a very high MAR index, and (III) genera/species that are easy to culture under laboratory conditions dominate in the qualitative and quantitative composition of the studied bacteria.     

Mathematical modeling of Microcystis aeruginosa growth and [D-Leu1] microcystin-LR production in culture media at different temperatures

The effect of temperature (26 °C, 28 °C, 30 °C and 35 °C) on the growth of native CAAT-3-2005 Microcystis aeruginosa and the production of Chlorophyll-a (Chl-a) and Microcystin-LR (MC-LR) were examined through laboratory studies. Kinetic parameters such as specific growth rate (μ), lag phase duration (LPD) and maximum population density (MPD) were determined by fitting the modified Gompertz equation to the M. aeruginosa strain cell count (cells mL⁻¹). A 4.8-fold increase in μ values and a 10.8-fold decrease in the LPD values were found for M. aeruginosa growth when the temperature changed from 15 °C to 35 °C. The activation energy of the specific growth rate (Eμ) and of the adaptation rate (E₁/LPD) were significantly correlated (R² = 0.86). The cardinal temperatures estimated by the modified Ratkowsky model were minimum temperature = 8.58 ± 2.34 °C, maximum temperature = 45.04 ± 1.35 °C and optimum temperature = 33.39 ± 0.55 °C.Maximum MC-LR production decreased 9.5-fold when the temperature was increased from 26 °C to 35 °C. The maximum production values were obtained at 26° C and the maximum depletion rate of intracellular MC-LR was observed at 30–35 °C. The MC-LR cell quota was higher at 26 and 28 °C (83 and 80 fg cell⁻¹, respectively) and the MC-LR Chl-a quota was similar at all the different temperatures (0.5–1.5 fg ng⁻¹).The Gompertz equation and dynamic model were found to be the most appropriate approaches to calculate M. aeruginosa growth and production of MC-LR, respectively. Given that toxin production decreased with increasing temperatures but growth increased, this study demonstrates that growth and toxin production processes are uncoupled in M. aeruginosa. These data and models may be useful to predict M. aeruginosa bloom formation in the environment.     

Development response of Spodoptera exigua to eight constant temperatures: Linear and nonlinear modeling

Temperature-dependent development of Spodoptera exigua (Hübner) were evaluated at eight constant temperatures of 12, 15, 20, 25, 30, 33, 34 and 36 °C with a variation of 0.5 °C on sugar beet leaves. No development occurred at 12 °C and 36 °C. Total developmental time varied from 120.50 days at 15 °C to 14.50 days at 33 °C. As temperature increased from 15 °C to 33 °C, developmental rate (1/developmental time) of S. exigua increased but declined at 34 °C. The lower temperature threshold (T_min) was estimated to be 12.98 °C and 12.45 °C, and the thermal constant (K) was 294.99 DD and 311.76 DD, using the traditional and Ikemoto–Takai linear models, respectively. The slopes of the Ikemoto–Takai linear model for different immature stages were different, violating the assumption of rate isomorphy. Data were fitted to three nonlinear models to predict the developmental rate and estimate the critical temperatures. The T_min values estimated by Lactin-2 (12.90 °C) and SSI (13.35 °C) were higher than the value estimated by Briere-2 (8.67 °C). The estimated fastest development temperatures (T_fast) by the Briere-2, Lactin-2 and SSI models for overall immature stages development of S. exigua were 33.4 °C, 33.9 °C and 32.4 °C, respectively. The intrinsic optimum temperature (T_Φ) estimated from the SSI model was 28.5 °C, in which the probability of enzyme being in its native state is maximal. The upper temperature threshold (T_max) values estimated by these three nonlinear models varied from 34.00 °C to 34.69 °C. These findings on thermal requirements can be used to predict the occurrence, number of generations and population dynamics of S. exigua.     

Knee joint angle affects EMG–force relationship in the vastus intermedius muscle

It is not understood how the knee joint angle affects the relationship between electromyography (EMG) and force of four individual quadriceps femoris (QF) muscles. The purpose of this study was to examine the effect of the knee joint angle on the EMG–force relationship of the four individual QF muscles, particularly the vastus intermedius (VI), during isometric knee extensions. Eleven healthy men performed 20–100% of maximal voluntary contraction (MVC) at knee joint angles of 90°, 120° and 150°. Surface EMG of the four QF synergists was recorded and normalized by the root mean square during MVC. The normalized EMG of the four QF synergists at a knee joint angle of 150° was significantly lower than that at 90° and 120° (P < 0.05). Comparing the normalized EMG among the four QF synergists, a significantly lower normalized EMG was observed in the VI at 150° as compared with the other three QF muscles (P < 0.05). These results suggest that the EMG–force relationship of the four QF synergists shifted downward at an extended knee joint angle of 150°. Furthermore, the neuromuscular activation of the VI was the most sensitive to change in muscle length among the four QF synergistic muscles.     

Effect of hydrothermolysis process conditions on pretreated switchgrass composition and ethanol yield by SSF with Kluyveromyces marxianus IMB4

Hot compressed liquid water was used to treat switchgrass in a method called hydrothermolysis to disrupt lignin, dissolve hemicellulose, and increase accessibility of cellulose to cellulase. Three temperatures (190, 200, and 210 °C) and hold times (10, 15, and 20 min) were tested. Switchgrass treated at 190 °C for 10 min had the greatest xylan recovery in the prehydrolyzate. Less than 0.65 g/L glucose were released into the prehydrolyzate for all pretreatment conditions, indicating most glucose was retained as cellulose in the solid substrate. 5-Hydroxymethylfurfural (HMF) and furfural formation in the prehydrolyzate were found to be less than 1 g/L for all treatments. The highest concentration of ethanol, 16.8 g/L (72% of theoretical), was produced from switchgrass pretreated at 210 °C and 15 min using simultaneous saccharification and fermentation (SSF) at 45 °C with the thermotolerant yeast Kluyveromyces marxianus IMB4 and 15 FPU cellulase/g glucan.