Screening,Molecular Cloning,and Biochemical Characterization of an Alcohol Dehydrogenase from Pichia pastoris Useful for the Kinetic Resolution of a Racemic β‐Hydroxy‐β‐trifluoromethyl Ketone

The stereoselective synthesis of chiral 1,3‐diols with the aid of biocatalysts is an attractive tool in organic chemistry. Besides the reduction of diketones, an alternative approach consists of the stereoselective reduction of β‐hydroxy ketones (aldols). Thus, we screened for an alcohol dehydrogenase (ADH) that would selectively reduce a β‐hydroxy‐β‐trifluoromethyl ketone. One potential starting material for this process is readily available by aldol addition of acetone to 2,2,2‐trifluoroacetophenone. Over 200 strains were screened, and only a few yeast strains showed stereoselective reduction activities. The enzyme responsible for the reduction of the β‐hydroxy‐β‐trifluoromethyl ketone was identified after purification and subsequent MALDI‐TOF mass spectrometric analysis. As a result, a new NADP⁺‐dependent ADH from Pichia pastoris (PPADH) was identified and confirmed to be capable of stereospecific and diastereoselective reduction of the β‐hydroxy‐β‐trifluoromethyl ketone to its corresponding 1,3‐diol. The gene encoding PPADH was cloned and heterologously expressed in Escherichia coli BL21(DE3). To determine the influence of an N‐ or C‐terminal His‐tag fusion, three different recombinant plasmids were constructed. Interestingly, the variant with the N‐terminal His‐tag showed the highest activity; consequently, this variant was purified and characterized. Kinetic parameters and the dependency of activity on pH and temperature were determined. PPADH shows a substrate preference for the reduction of linear and branched aliphatic aldehydes. Surprisingly, the enzyme shows no comparable activity towards ketones other than the β‐hydroxy‐β‐trifluoromethyl ketone.     

Comparison of Soybean and Cottonseed Oils upon Hydrogenation with Nickel,Palladium and Platinum Catalysts

There is current interest in reducing the trans fatty acids (TFA) in hydrogenated vegetable oils because consumption of foods high in TFA has been linked to increased serum cholesterol content. In the interest of understanding the TFA levels, hydrogenation was carried out in this work on soybean oil and cottonseed oil at two pressures (2 and 5 bar) and 100 °C using commercially available Ni, Pd, and Pt catalysts. The TFA levels and the fatty acid profiles were analyzed by gas chromatography. The iodine value of interest is ~70 for all-purpose shortening and 95–110 for pourable oil applications. In all cases, higher hydrogen pressures produced lower levels of TFA. In the range of 70–95 iodine values for the hydrogenated products, the Pt catalyst gave the least TFA, followed closely by Ni, and then Pd, for both oils. For all three catalysts at 2- and 5-bar pressures and 70–95 iodine values, cottonseed oil contained noticeably less TFA than soybean oil; this is probably because cottonseed oil contains a lower total amount of olefin-containing fatty acids relative to soybean oil. Approximate kinetic modeling was also done on the hydrogenation data that provided additional confirmation of data consistency.     

Diazoverbindungen. 72. (Diazoalkyl)phosphane – Synthese durch elektrophile Diazoalkansubstitution und oxidative Additionsreaktionen am Phosphor

Diazo Compounds. 72. Diazoalkylphosphanes – Synthesis by Electrophilic Diazoalkane Substitution and Oxidative Addition Reactions at Phosphorus Electrophilic diazoalkane substitution of the diazomethyl compounds 1a,b with the chloro phosphanes 2a-o in the presence of lithium diethylamide yields the diazoalkyl phosphanes 3a-z . Oxidative addition of oxygen, sulfur and selenium at phosphorus leads into the series of oxo, thioxo and selenoxo phosphanes having diazoalkyl substituents ( 4a-d, 5a-m and 7a-d ). The silyl group of 5n,o is cleaved by chromatography on aluminium oxide to yield the (diazomethyl)phosphane sulfides 6a,b .     

Starch-Derived Products for Detergents

Recent developments in the formulation of detergents have been driven by a strong consumer demand for natural and biodegradable products. Detergent manufacturers responded to this demand with corresponding products and advertising slogans such as “fully biodegradable”, “natural” or even “double natural” to oust their competitors. In a detergent formulation, starch- derived products can in principle be used for the following purposes: as the hydrophilic head group in surfactants, as the starting material for (poly)carboxylate co-builders and as the backbone of bleaching activators. Non-ionic classical surfactants can be replaced by alkylpolyglucosides (APGs), a class of products completely based on renewable resources such as glucose and fatty alcohols derived from natural fatty acids. Sodium tripolyphosphate (STPP), the product responsible for the eutrophication of surface waters, can be substituted by a combination of an inorganic zeolite and highly oxidised starch (dicarboxylic starch) or by citrate. Acetylated polyols derived from hydrogenated carbohydrates such as sorbitol can take over the function of the petrochemically-based tetraacetyl ethylene diamine (TAED) used as activator to allow perborate bleaching at lower washing temperatures.     

Erythritol,a New Raw Material for Food and Non-food Applications

In response to an increasing demand from consumers for healthier and calorie controlled foods, Cerestar has developed a new food ingredient, erythritol. Erythritol can be produced from starch by a full biotechnological process, combining enzymatic and fermentative conversions. The use of an osmophilic yeast allows the fermentation step to be performed at high dry substance, giving an economic advantage. An extremely pure end product is then easily obtained by final crystallisation. Erythritol is structurally a polyol and shares the health properties of other polyols such as being toothfriendly and safe for diabetics. However, it offers in addition two very important nutritional advantages: a lower calorific value (0.3 Kcal/g) and a good tolerance. This is due to its low molecular weight, which allows erythritol to be rapidly absorbed from the small intestine, with subsequent excretion in the urine. Fermentation in the colon is therefore excluded and any resulting gastro-intestinal discomfort avoided. The combination of these properties makes erythritol a unique low calorie bulk sweetener. From a functionality point of view, erythritol is a moderately sweet bulking agent with a cooling taste: workability is similar to other polyols. It has a taste profile close to that of sucrose and may therefore improve the taste quality of a blend with intense sweeteners. Its low solubility and ease of crystallisation, make erythritol very suitable for applications which require a crystalline sweetener, such as chocolate. Other potential application areas are bakery, table-top and confectionery. Presently erythritol is under evaluation to establish beyond any doubt its safety and to obtain food approval as a new, low calorie, bulk sweetener. Potential non-food applications of erythritol are in polymers, fine chemicals and pharmaceutical intermediates.     

卡车司机对于车载技术提供反馈的认知研究

利用车载技术为卡车司机提供反馈是帮助卡车司机提高安全性的一种方法.使用焦点群体访谈的方法对大量卡车司机及运输行业相关人员(管理人员和交警)进行了调研,以探索卡车司机对于车载技术提供反馈的认知.访谈涉及驾驶中的安全和不安全行为、车载技术提供反馈的最好方式、使用车载技术提供反馈的优缺点等.为开发相应的技术提供了有益的建议,并有利于进一步探索卡车司机对于车载技术提供反馈的认知.