Construction of a plasmid carrying both CTP synthetase and a fused gene formed from cholinephosphate cytidylyltransferase and choline kinase genes and its application to industrial CDP-choline production: enzymatic production of CDP-choline from orotic acid (Part II)

A new method for enzymatic production of cytidine diphosphate choline (CDP-choline) from orotic acid and choline chloride was developed. To establish an industrial manufacturing process, we constructed a plasmid, pCKG55, which simultaneously expressed in Escherichia coli the three following enzymes; CTP synthetase (encoded by the pyrG gene from E. coli), cholinephosphate cytidylyltransferase (encoded by the CCT gene from Saccharomyces cerevisiae), and choline kinase (encoded by the CKI gene from S. cerevisiae). CCT and CKI genes on pCKG55 were designed to be expressed as a single CCT/CKI fused protein. This CCT/CKI fused protein retained both activities and the thermal stability of its cholinephosphate cytidylyltransferase activity was nearly the same as the native CCT enzyme. Corynebacterium ammoniagenes KY13505 and E. coli MM294/pCKG55 were cultured in 5-liter jar fermentor independently. Equal volumes of each broth were mixed in a 2-liter jar fermentor, and then the enzymatic reaction was done using 47 mM orotic acid and 60 mM choline chloride as substrates. After 23 h of the reaction at 32 degrees C, 21.5 mM (11 g/liter) of CDP-choline was accumulated.     

A 160-kDa Protein Is Essential for Hemocyanin-derived Melanosis of Prawn

ABSTRACT A purified 160-kDa protein, isolated from the cuticle of kurama prawn and named melanosis collaborating factor (MCF), was a key factor in promoting melanosis by a cooperative reaction with hemocyanin-derived phenoloxidase but hemocyanin itself was incapable of producing black pigment. The enzymatic activity of MCF in the body of the prawn was very stable against the process of freezing and thawing; the enzyme maintained more than 80% of its activity after 3 mo of storage at −2 5 °C. These results indicated that MCF, acting in conjunction with hemocyanin-derived phenoloxidase, played a crucial role in postharvest melanogenesis in prawn.     

Selective hydrogenation of oleic acid to 9-octadecen-1-ol: Catalyst preparation and optimum reaction conditions

A new catalyst, ruthenium-tin-alumina is found to selectively hydrogenate oleic acid to 9-octadecen-1-ol (oleyl + elaidyl alcohol) at low pressure with high yield. Catalyst preparation methods, catalyst raw materials and activation conditions have a significant effect on the activity of the catalyst. The optimum atomic ratio of ruthenium to tin is about 1:2. Catalyst prepared by an improved sol-gel method shows higher activity and selectivity than catalysts prepared by impregnation and coprecipitation methods. Chloride is found to have a negative effect on catalytic activity. The best catalyst is prepared from chloride-free ruthenium and tin raw materials. Under the optimum reaction conditions of 250°C and 5.6 MPa, the selectivities for 9-octadecen-1-ol and total alcohol (9-octadecen-1-ol + stearyl alcohol) formation are 80.9% and 97%, respectively, at a conversion of 81.3%.     

Applications of the 4×4 determinant method and the POLYGON ENGINE

In the first half of the paper, various types of processing pertaining to a polygon, using the 4×4 determinant theories are explained along with a new containment test algorithm of a point in a polygon. In the latter half of the paper, a general-purpose geometric processor, the POLYGON ENGINE, is presented which can deal with various types of interference problems, such as Boolean operations in solid modelling, hidden line and surface eliminations, ray tracing and so on. It is, a successor of the TRIANGLE PROCESSOR and is also based upon the 4×4 determinant theories [4–6]. While the TRIANGLE PROCESSOR processes a triangulated polygon on a triangle-by-triangle basis, the POLYGON ENGINE can treat a polygon without triangulation. The latter is expected to be more functional, more efficient and easier to use.     

N，O，O′─三（对甲苯磺酰基）双（2─羟乙基）胺的合成

二乙醇胺和对甲苯磺酰氯在三乙胺存在下，反应生成Ｎ，０，Ｏ′－三（对甲苯磺酰基）双（２－羟乙基）胺，产率９３％。从反应混合物中分离得副产物Ｎ，Ｏ－二（对甲苯磺酰基）双（２－羟乙基）胺。     

Transient analysis of the release and reduction of NOx using a Pt/Ba/Al2O3 catalyst

The release and reduction of NO_x in a NO_x storage-reduction (NSR) catalyst were studied with a transient reaction analysis in the millisecond range, which was made possible by the combination of pulsed injection of gases and time resolved time-of-flight mass spectrometry. After an O₂ pulse and a subsequent NO pulse were injected into a pellet of the Pt/Ba/Al₂O₃ catalyst, the time profiles of several gas products, NO, N₂, NH₃ and H₂O, were obtained as a result of the release and reduction of NO_x caused by H₂ injection. Comparing the time profiles in another analysis, which were obtained using a model catalyst consisting of a flat 5 nmPt/Ba(NO₃)₂/cordierite plate, the release and reduction of NO_x on Pt/Ba/Al₂O₃ catalyst that stored NO_x took the following two steps; in the first step NO molecules were released from Ba and in the second step the released NO was reduced into N₂ by H₂ pulse injection. When this H₂ pulse was injected in a large amount, NO was reduced to NH₃ instead of N₂.

A only small amount of H₂O was detected because of the strong affinity for alumina support. We can analyze the NO_x regeneration process to separate two steps of the NO_x release and reduction by a detailed analysis of the time profiles using a two-step reaction model. From the result of the analysis, it is found that the rate constant for NO_x release increased as temperature increase.     

A pilot survey of 39 Victorian WWTP effluents using a high speed luminescent umu test in conjunction with a novel GC-MS-database technique for automatic identification of micropollutants

In 2007, samples of treated effluent were collected at point of discharge to the environment from 39 wastewater treatment plants (WWTPs) located across Victoria, Australia grouped by treatment type. Sample genotoxicity was assessed with a high-throughput luminescent umu test method using Salmonella typhimurium TL210 strain, with and without addition of a commercially available metabolic activation system. Samples were also screened using a gas chromatographic-mass spectrometric mass-structure database recognition method. A genotoxic response was observed in half of the samples tested without metabolic activation system (相似文献   

Molecular Depolymerization of Moist Sweet Potato Starch Melt by Shearing Force under an Elevated Temperature

Sweet potato starch (SPS) with 20 or 25% moisture was heated and then fused at 150°C for 15 min under pressurization at 30–500 Kg/cm². At the same time, the heated SPS melt was extruded through a capillary tube under specified pressure in order to give shearing force. Thus, intact SPS, heated SPS and heated sheared SPS specimen were prepared and examined. The heated and heated sheared SPS did not contain any pyrolysis products because their spectrum profiles were the same as that of intact SPS. By heating and shearing, the intact SPS molecule was depolymerized into small molecules including a few amount of glucose. This finding was revealed by means of molecular weight estimation by viscometry and by gel filtration. The shearing toward SPS melt markedly stimulated the depolymerization while the effect of only heating on depolymerization of SPS was modulate in comparison with heated shearing. Water solubilities of intact SPS, heated SPS and heated sheared SPS with 20% moisture were 0.07, 3.5 and 51.5% respectively, and also the degrees of gelatinization of 1.8% for intact starch increased markedly to 29% for heated SPS and 80–95% (depending on the pressure at extrusion) for heated sheared SPS.