Young Hoon Jung

Dilute acid pretreatment of lignocellulose for whole slurry ethanol fermentation.

Dilute sulfuric acid pretreatment of oil palm empty fruit bunches (EFB) followed by the whole slurry fermentation of the pretreated EFB slurry was investigated. The optimized pretreatment conditions were at 1% (w/v) sulfuric acid with 3 min ramping to 190 °C in a microwave digester. Pretreated and washed EFB exhibited enzymatic digestibility of 88.5% of theoretical glucose yield after 48 h of hydrolysis. When the whole slurry of pretreated and neutralized EFB was used in simultaneous saccharification and fermentation (SSF) using cellulase and Saccharomyces cerevisiae, sulfuric acid-pretreated EFB resulted in 52.5% of theoretical ethanol yield based on total glucan in the untreated initial EFB after 72 h of SSF. When pretreated EFB slurry was treated with activated carbon before subjecting to SSF, the SSF furnished 87.5% ethanol yield based on the initial glucan content in untreated EFB (after 48 h of SSF).

Global Metabolite Profiling of Synovial Fluid for the Specific Diagnosis of Rheumatoid Arthritis from Other Inflammatory Arthritis

Currently, reliable biomarkers that can be used to distinguish rheumatoid arthritis (RA) from other inflammatory diseases are unavailable. To find possible distinctive metabolic patterns and biomarker candidates for RA, we performed global metabolite profiling of synovial fluid samples. Synovial fluid samples from 38 patients with RA, ankylosing spondylitis, Behçets disease, and gout were analyzed by gas chromatography/time-of-flight mass spectrometry (GC/TOF MS). Orthogonal partial least-squares discriminant and hierarchical clustering analyses were performed for the discrimination of RA and non-RA groups. Variable importance for projection values were determined, and the Wilcoxon-Mann-Whitney test and the breakdown and one-way analysis of variance were conducted to identify potential biomarkers for RA. A total of 105 metabolites were identified from synovial fluid samples. The score plot of orthogonal partial least squares discriminant analysis showed significant discrimination between the RA and non-RA groups. The 20 metabolites, including citrulline, succinate, glutamine, octadecanol, isopalmitic acid, and glycerol, were identified as potential biomarkers for RA. These metabolites were found to be associated with the urea and TCA cycles as well as fatty acid and amino acid metabolism. The metabolomic analysis results demonstrated that global metabolite profiling by GC/TOF MS might be a useful tool for the effective diagnosis and further understanding of RA.

Aqueous ammonia pretreatment of oil palm empty fruit bunches for ethanol production

Oil palm empty fruit bunches (EFB) were pretreated by aqueous ammonia soaking for ethanol production. Pretreated EFB, which were pretreated at the optimal conditions of 60 °C, 12 h, and 21% (w/w) aqueous ammonia, showed 19.5% and 41.4% glucose yields during an enzymatic digestibility test for 96 h when using 15 and 60 FPU of cellulase, respectively. Using the pretreated EFB, simultaneous saccharification and fermentation for 168 h with 5% (w/v) glucan loading and 60 FPU of cellulase and 30 CBU of β-glucosidase per gram glucan resulted in ethanol production of 18.6 g/L titer, 65.6% of theoretical maximum yield, and 0.11 g/L/h of productivity.

Ethanol production from oil palm trunks treated with aqueous ammonia and cellulase

Oil palm trunks are a possible lignocellulosic source for ethanol production. Low enzymatic digestibility of this type of material (11.9% of the theoretical glucose yield) makes pretreatment necessary. An enzymatic digestibility of 95.4% with insoluble solids recovery of 49.8% was achieved after soaking shredded oil palm trunks in ammonia under optimum conditions (80°C, 1:12 solid-to-liquid ratio, 8h and 7% (w/w) ammonia solution). Treatment with 60 FPU of commercial cellulase (Accellerase 1000) per gram of glucan and fermentation with Saccharomyces cerevisiae D(5)A resulted in an ethanol concentration of 13.3g/L and an ethanol yield of 78.3% (based on the theoretical maximum) after 96 h. These results indicate that oil palm trunks are a biomass feedstock that can be used for bioethanol production.

Mimicking the Fenton reaction-induced wood decay by fungi for pretreatment of lignocellulose.

In this study, the Fenton reaction, which is naturally used by fungi for wood decay, was employed to pretreat rice straw and increase the enzymatic digestibility for the saccharification of lignocellulosic biomass. Using an optimized Fentons reagent (FeCl3 and H2O2) for pretreatment, an enzymatic digestibility that was 93.2% of the theoretical glucose yield was obtained. This is the first report of the application of the Fenton reaction to lignocellulose pretreatment at a moderate temperature (i.e., 25°C) and with a relatively high loading of biomass (i.e., 10% (w/v)). Substantial improvement in the process economics of cellulosic fuel and chemical production can be achieved by replacing the conventional pretreatment with this Fenton-mimicking process.

Evaluation of a transgenic poplar as a potential biomass crop for biofuel production

A transgenic poplar, in which the RabG3bCA gene from Arabidopsis was overexpressed, was analyzed for its biomass composition and enzymatic digestibility after chemical pretreatment. In comparison with a wild-type poplar (WT), the transgenic poplar (OX8) showed 9.8% higher glucan content. The levels of other biomass components did not differ greatly between WT and OX8. When WT and OX8 samples were pretreated by sulfuric acid (1%, w/v at 190 °C), sodium hydroxide (1%, w/v at 190 °C), or ammonia (14%, w/w at 80 °C), the washed pretreated solids of OX8 exhibited a higher enzymatic digestibility than those of WT in each chemical pretreatment. The sodium hydroxide pretreatment was the most effective among the three pretreatment processes, showing 58.7% and 69.4% of theoretical glucose yield from the saccharification of pretreated OX8 and WT, respectively. The transgenic poplar, growing faster and taller, was found to contain more glucan and have a higher enzymatic digestibility than WT.

One-pot pretreatment, saccharification and ethanol fermentation of lignocellulose based on acid–base mixture pretreatment

Currently, for the production of cellulosic ethanol, multi-step unit operations, including pretreatment, solid/liquid (S/L) separation, solids washing, liquid detoxification, neutralization, enzymatic hydrolysis and fermentation, are the commonly required steps responsible for elevating the capital and operating costs. To simplify these steps, consolidated bioprocessing (CBP), focusing on the multi-functional microbial strains, was proposed. However, this process has not been commercialized yet. In this study, using an acid–base mixture as a pretreatment catalyst, pretreatment, saccharification and fermentation were performed in one pot without S/L separation, neutralization and detoxification. From the one-pot process based on the acid–base mixture pretreatment (190 °C, 2 min and 0.15 (w/v) acid–base mixture) and 15 FPU of cellulase per g glucan and Sacchromyces cerevisiae, 70.7% of the theoretical maximum ethanol yield (based on the initial amount of glucan in the untreated rice straw) was obtained. This was comparable to the estimated ethanol yield of 72.9%, assuming a 90% glucan recovery yield after pretreatment × a 90% glucose yield from saccharification × a 90% ethanol yield from ethanol fermentation performed in three separate pots. These results suggest that the entire slurry processing of lignocellulose in one pot could be an attractive way to achieve economic sustainability in the production of fuel from lignocellulose.

Combination of high solids loading pretreatment and ethanol fermentation of whole slurry of pretreated rice straw to obtain high ethanol titers and yields.

In cellulosic ethanol production using lignocellulose, an increase in biomass solids loading during the pretreatment process significantly affects the final ethanol titer and the production cost. In this study, pretreatment using rice straw at high solids loading (20% (w/v)) was evaluated, using maleic acid as a catalyst. After pretreatment at optimal conditions of 190°C, 20 min, and 0.2% or 5% (w/v) maleic acid, the highest enzymatic digestibility obtained was over 80%. Simultaneous saccharification and fermentation (SSF) of the whole slurry of pretreated rice straw in the presence of activated carbon to separate inhibitory compounds generated a high ethanol yield of 62.8%, based on the initial glucan in unpretreated rice straw. These findings suggest that high solids loading pretreatment using maleic acid and SSF of the whole slurry of pretreated rice straw can be combined to improve the process economics of ethanol production.

Customized optimization of cellulase mixtures for differently pretreated rice straw

Lignocellulose contains a large amount of cellulose but is recalcitrant to enzymatic hydrolysis, which yields sugars for fuels or chemicals. Various pretreatment methods are used to improve the enzymatic digestibility of cellulose in lignocellulose. Depending on the lignocellulose types and pretreatment methods, biomass compositions and physical properties significantly vary. Therefore, customized enzyme mixtures have to be employed for the efficient hydrolysis of pretreated lignocellulose. Here, using three recombinant model enzymes consisting of endoglucanase, cellobiohydrolase, and xylanase with a fixed amount of β-glucosidase, the optimal formulation of enzyme mixtures was designed for two differently pretreated rice straws (acid-pretreated or alkali-pretreated rice straw) by the mixture design methodology. As a result, different optimal compositions for the enzyme mixtures were employed depending on the type of pretreatment of rice straw. These results suggest that customized enzyme mixtures for pretreated lignocellulosic biomass are necessary to obtain increased sugar yields and should be considered in the industrial utilization of lignocellulose.

Atmospheric vs. anaerobic processing of metabolome samples for the metabolite profiling of a strict anaerobic bacterium, Clostridium acetobutylicum

Well‐established metabolome sample preparation is a prerequisite for reliable metabolomic data. For metabolome sampling of a Gram‐positive strict anaerobe, Clostridium acetobutylicum, fast filtration and metabolite extraction with acetonitrile/methanol/water (2:2:1, v/v) at −20°C under anaerobic conditions has been commonly used. This anaerobic metabolite processing method is laborious and time‐consuming since it is conducted in an anaerobic chamber. Also, there have not been any systematic method evaluation and development of metabolome sample preparation for strict anaerobes and Gram‐positive bacteria. In this study, metabolome sampling and extraction methods were rigorously evaluated and optimized for C. acetobutylicum by using gas chromatography/time‐of‐flight mass spectrometry‐based metabolomics, in which a total of 116 metabolites were identified. When comparing the atmospheric (i.e., in air) and anaerobic (i.e., in an anaerobic chamber) processing of metabolome sample preparation, there was no significant difference in the quality and quantity of the metabolomic data. For metabolite extraction, pure methanol at −20°C was a better solvent than acetonitrile/methanol/water (2:2:1, v/v/v) at −20°C that is frequently used for C. acetobutylicum, and metabolite profiles were significantly different depending on extraction solvents. This is the first evaluation of metabolite sample preparation under aerobic processing conditions for an anaerobe. This method could be applied conveniently, efficiently, and reliably to metabolome analysis for strict anaerobes in air. Biotechnol. Bioeng. 2014;111: 2528–2536.