The ease of genetic manipulation, low cost, rapid growth and number of previous studies have made Escherichia coli one of the most widely used microorganism species for producing recombinant proteins. In this post-genomic era, challenges remain to rapidly express and purify large numbers of proteins for academic and commercial purposes in a high-throughput manner. In this review, we describe several state-of-the-art approaches that are suitable for the cloning, expression and purification, conducted in parallel, of numerous molecules, and we discuss recent progress related to soluble protein expression, mRNA folding, fusion tags, post-translational modification and production of membrane proteins. Moreover, we address the ongoing efforts to overcome various challenges faced in protein expression in E. coli, which could lead to an improvement of the current system from trial and error to a predictable and rational design.

To isolate bacteria responsible for the biodegradation of naphthalene, BTEX (benzene, toluene, ethylbenzene, and o-, m-, and p-xylene), and aliphatic hydrocarbons in petroleum-contaminated soil, three enrichment cultures were established using soil extract as the medium supplemented with naphthalene, BTEX, or n-hexadecane. Community analyses showed that Paraburkholderia species were predominant in naphthalene and BTEX, but relatively minor in n-hexadecane. Paraburkholderia aromaticivorans BN5 was able to degrade naphthalene and all BTEX compounds, but not n-hexadecane. The genome of strain BN5 harbors genes encoding 29 monooxygenases including two alkane 1-monooxygenases and 54 dioxygenases, indicating that strain BN5 has versatile metabolic capabilities, for diverse organic compounds: the ability of strain BN5 to degrade short chain aliphatic hydrocarbons was verified experimentally. The biodegradation pathways of naphthalene and BTEX compounds were bioinformatically predicted and verified experimentally through the analysis of their metabolic intermediates. Some genomic features including the encoding of the biodegradation genes on a plasmid and the low sequence homologies of biodegradation-related genes suggest that biodegradation potentials of strain BN5 may have been acquired via horizontal gene transfers and/or gene duplication, resulting in enhanced ecological fitness by enabling strain BN5 to degrade all compounds including naphthalene, BTEX, and short aliphatic hydrocarbons in contaminated soil.

In this study, two experiments were conducted to evaluate the total mixed ration with fermented feed (TMRF) and total mixed ration (TMR) by rumen in vitro fermentation and their effects on the growth performance and blood characteristics of Hanwoo steers. In experiment 1, three Hanwoo steers (600±47 kg), each permanently fitted with a ruminal cannula were used. In this experiment, three diets designated as T1, TMRF (18.4% fermented feed, tall fescue, mammoth wild rye forage and whole crop barley); T2, TMRF (17.7% fermented feed, rice straw and whole crop barley); and T3, TMR (rice straw, whole crop barley and probiotics, but no fermented feed), which were subjected to rumen in vitro fermentation for 48 h. The results demonstrated that DM disappearance rate gradually increased with advancing fermentation time, but T1 and T2 were higher than the T3 (p<0.05) from 3 h to 12 h, but insignificant (p>0.05) at 24 and 48 h. None of the specific VFAs were affected except for acetic and non volatile lactic acids, which were produced more in T2 than in T1 and T3 at 24 h and 48 h of incubation. A/P was lower in T1 and T2 than inT3 at 24 h (p<0.05) and 48 h (p>0.05) of incubation. These results confirmed that TMRF-related treatment shows a superior performance to that of TMR during the ruminal fermentation period. In experiment 2, the three diets in experiment 1 plus 1 more control diet (concentrates, probiotics and 2% rice straw of body weight) were fed to the 48 Hanwoo steers (160±10 kg) for a period of 168 d. The results demonstrated that the daily and total live weight gain and feed efficiency were higher (p<0.05) in the TMRF and TMR groups than in the control group. SGOT, SGPT and BUN (p<0.05) were reduced in TMRF relative to the control and TMR groups by 168 d which confirmed that TMRF shows better blood profiles than the TMR and control groups. Overall, these results appear to show that TMRF has better in vitro ruminal characteristics than those of TMR; growth performance and blood profiles were also found to be superior in TMRF than in the TMR and control groups. Thus, our findings suggest that TMRF-based feed supplies are favorable for Hanwoo cattle.

Ionic liquids (ILs) are low melting organic salts that potentially comprise wide application due to their fascinating properties and have emerged as promising "green" replacements for volatile organic solvents. Despite their nonmeasurable vapor pressure, some quantities of ILs will soon be present in effluent discharges since they do have significant solubility in water. Recently, the toxic effects of ILs toward aquatic communities have been intensively investigated, but little information is available concerning the biodegradable properties of these compounds. The objective of this study was to identify the metabolites generated during the biotransformation of 1-butyl-3-methylpyridinium by microorganisms in aerobic activated sludge. The obtained results revealed that the alkylpyridinium salt was metabolized through the sequential oxidization in different positions of the alkyl side chains. High-performance liquid chromatography and mass-spectrometry analyses demonstrated that this biodegradation led to the formation of 1-hydroxybutyl-3-methylpyridinium, 1-(2-hydroxybutal)-3-methylpyridinium, 1-(2-hydroxyethyl)-3-methylpyridinium, and methylpyridine. On the basis of these intermediate products, biodegradation pathways were also suggested. These findings provide the basic information that might be useful for assessing the factors related to the environmental fate and behavior of this commonly used pyridinium IL.