两相系统在疏水性有机污染物高效降解菌富集培养中的应用研究进展

环境中疏水性有机污染物(Hydrophobic organic pollutants,HOPs)的浓度日益增加,获取HOPs高效降解功能微生物能有效提高HOPs污染治理效率。近年来,利用两相系统促进HOPs微生物降解转化的研究已取得一定进展。本文重点从两相系统的结构特点及其富集HOPs降解功能微生物的原理、主要影响因素和应用情况等方面进行综述,并在此基础上对两相系统在毒害性HOPs微生物加速降解脱毒中所存在的主要问题及其应用前景进行讨论和展望。     

Recombinant carbazole-degrading strains for enhanced petroleum processing

Biotechnological upgrading of fossil fuels is of increasing interest as remaining stocks of petroleum show increasing levels of contaminants such as heavy metals, sulfur and nitrogen-containing heteroaromatic compounds. Carbazole is of particular interest as a major petroleum component known to reduce refining yields through catalyst poisoning. In this study, the biotransformation of carbazole was successfully demonstrated in a liquid two-phase system, when solubilized in either 1-methylnaphthalene or in diesel fuel. The effects of solvent toxicity were investigated by expressing the carbazole-transformation genes from MB1332, a rifampicin-resistant derivative of Pseudomonas sp. LD2, in a solvent-resistant heterologous host, P. putida Idaho [1]. This solvent-resistant strain successfully degraded carbazole solubilized in 1-methylnaphthalene and in the presence of 10 vol% xylenes similar to the non-recombinant strain Pseudomonas sp. LD2. Identification of a suitable recombinant host, however, was essential for further investigations of partial pathway transformations. Recombinant P. putida Idaho expressing only the initial dioxygenase enzymes transformed carbazole to an intermediate well retained in the oil phase. Partial carbazole transformation converts carbazole to non-aromatic species; their effect is unknown on refinery catalyst poisoning, but would allow almost complete retention of carbon content and fuel value. Electronic Publication     

Optimization of two-phase thermophilic anaerobic digestion of biowaste for hydrogen and methane production through reject water recirculation

The optimization of a two-phase thermophilic anaerobic process treating biowaste for hydrogen and methane production was carried out at pilot scale using two stirred reactors (CSTRs) and without any physical/chemical pre-treatment of inoculum. During the experiment the hydrogen production at low hydraulic retention time (3d) was tested, both with and without reject water recirculation and at two organic loading rate (16 and 21 kgTVS/m³d). The better yields were obtained with recirculation where the pH reached an optimal value (5.5) thanks to the buffering capacity of the recycle stream. The specific gas production of the first reactor was 51 l/kgVS_fed and H₂ content in biogas 37%. The mixture of gas obtained from the two reactors met the standards for the biohythane mix only when lower loading rate were applied to the first reactor, with a composition of 6.7% H₂, 40.1% CO₂ and 52.3% CH₄ the overall SGP being 0.78 m³/kgVS_fed.     

On a supplemented case-control design

The supplemented case-control design consists of a case-control sample and of an additional sample of disease-free subjects who arise from a given stratum of one of the measured exposures in the case-control study. The supplemental data might, for example, arise from a population survey conducted independently of the case-control study. This design improves precision of estimates of main effects and especially of joint exposures, particularly when joint exposures are uncommon and the prevalence of one of the exposures is low. We first present a pseudo-likelihood estimator (PLE) that is easy to compute. We further adapt two-phase design methods to find maximum likelihood estimates (MLEs) for the log odds ratios for this design and derive asymptotic variance estimators that appropriately account for the differences in sampling schemes of this design from that of the traditional two-phase design. As an illustration of our design we present a study that was conducted to assess the influence to joint exposure of hepatitis-B virus (HBV) and hepatitis-C virus (HCV) infection on the risk of hepatocellular carcinoma in data from Qidong County, Jiangsu Province, China.     

Improved (<Emphasis Type="Italic">R</Emphasis>)-phenylacetylcarbinol production with <Emphasis Type="Italic">Candida utilis</Emphasis> pyruvate decarboxylase at decreased organic to aqueous phase volume ratios

The effect of decreasing the organic (octanol) to aqueous phase volume ratio was evaluated in a two-phase enzymatic process for (R)-phenylacetylcarbinol (PAC) production. Decreasing the ratio from 1:1 to 0.43:1 at 4°C increased PAC in the organic phase from 112 g/l to 183 g/l with a 10% improvement in overall productivity. Interestingly, the rate of enzyme (pyruvate decarboxylase) activity loss was unaffected by the reduced phase ratio over the reaction period (48 h). At 20°C and 0.43:1 phase ratio the organic phase PAC concentration increased to 212 g/l and the overall productivity increased by 30% although the PAC yield (based on pyruvate) declined by about 10% due to greater byproduct acetoin formation at the higher temperature. Product recovery in such a system is facilitated both by the higher PAC concentration and the reduced organic phase volume.     

Optimization of lipase-catalyzed glucose fatty acid ester synthesis in a two-phase system containing ionic liquids and t-BuOH

Selective lipase-catalyzed synthesis of glucose fatty acid esters in two-phase systems consisting of an ionic liquid (1-butyl-3-methyl imidazolium tetrafluoroborate [BMIM][BF₄] or 1-butyl-3-methyl imidazolium hexafluorophosphate [BMIM][PF₆]) and t-butanol as organic solvent was investigated. The best enzyme was commercially available lipase B from Candida antarctica (CAL-B), but also lipase from Thermomyces lanuginosa (TLL) gave good conversion. After thorough optimization of several reaction conditions (chain-length and type of acyl donor, temperature, reaction time, percentage of co-solvent) conversions up to 60% could be achieved using fatty acid vinyl ester as acyl donors in [BMIM][PF₆] in the presence of 40% t-BuOH with CAL-B at 60 °C.     

Whole cell microbial transformation in cloud point system

Cloud point system, consisting of nonionic surfactant in an aqueous solution, has been developed as a novel medium for whole cell microbial transformation. The basic properties of cloud point system including phase separation and solubilization are introduced. The application of cloud point system for extractive microbial transformation is different from that of water-organic solvent two-phase partitioning system or aqueous two-phase system are discussed, which mainly focus on the biocompatibility of microorganism in a cloud point system and a downstream process of microbial transformation in cloud point system with oil-water-surfactant microemulsion liquid-liquid extraction for surfactant recovery and product separation. Finally, examples of whole cell microbial transformation in cloud point systems, especially in situ extraction of moderate polar substrate/product, are also presented.     

Modeling non-homologous end joining

Non-homologous end joining (NHEJ) is an important DNA repair pathway for DNA double-strand breaks. Several proteins, including Ku, DNA-PKcs, Artemis, XRCC4/Ligase IV and XLF, are involved in the NHEJ for the DNA damage detection, DNA free end processing and ligation. The classical model of NHEJ is a sequential model in which DNA-PKcs is first recruited by the Ku bound DNA prior to any other repair proteins. Recent experimental study ( [McElhinny et al., 2000], [Costantini et al., 2007], [17] and [Yano and Chen, 2008]) suggested that the recruitment ordering is not crucial. In this work, by proposing a mathematical model in terms of biochemical reaction network and performing stability and related analysis, we demonstrate theoretically that if DSB repair pathway independent of DNA-PKcs exists, then the classical sequential model and new two-phase model are essentially indistinguishable in the sense that DSB can be repaired thoroughly in both models when the repair proteins are sufficient.     

Reaction Medium Selection for An Enzymatic Peptide Synthesis in An Aqueous-Organic Two-Phase System

Efficient development of enzymatic synthesis in two-phase systems is closely related with appropriate selection of the reaction medium (especially the solvent and phase ratio). A selection procedure based on the calculation of the theoretically allowable conversion and product concentration is presented and applied to a peptide synthesis using papain. For the synthesis of the dipeptide Boc-Gly-Phe-OMe, the operating conditions have been determined, and the two-phase system to be used has been successfully selected (with trichloroethylene being the best solvent).     

Whole cell yeast biotransformations in two-phase systems: Effect of solvent on product formation and cell structure

Summary Biotransformation of benzaldehyde and pyruvate to (R)-phenylacetyl carbinol bySaccharomyces cerevisiae was investigated in two-phase aqueous-organic reaction media. With hexane as organic solvent, maximum biotransformation activity was observed with a moisture content of 10%. Of the organic solvents tested, highest biotransformation activities were observed with hexane and hexadecane, and lowest activities occurred with chloroform and toluene. Biocatalyst samples from biphasic media containing hexane, decane and toluene manifested no apparent cell structural damage when examined using scanning electron microscopy. In contrast, cellular biocatalyst recovered from two-phase systems containing chloroform, butylacetate and ethylacetate exhibited damage in the form of cell puncturing after different incubation periods. Phospholipids were detected in reaction media from biocatalytic systems which exhibited cell damage in electron micrographs. Phospholipid release was much lower in the two-phase systems containing toluene or hexane or in 100% aqueous biocatalytic system.