Evaluation of Kluyveromyces marxianus FII 510700 grown on a lactose-based medium as a source of a natural bioemulsifier

Mannoprotein with emulsification properties was extracted from the cell walls of Kluyveromyces marxianus grown on a lactose-based medium by autoclaving cells in a citrate buffer at pH 7.The purified product was evaluated for chemical and physical stability to establish its potential use as a natural emulsifier in processed foods. The yield of purified bioemulsifier from this strain of K. marxianus was 4–7% of the original dry cell weight. The purified product, at a concentration of 12 g l^–1, formed emulsions that were stable for 3 months when subjected to a range of pH (3–11) and NaCl concentrations (2–50 g l^–1). The composition of this mannoprotein was 90% carbohydrate (mannan) and 4–6% protein. These values are similar to mannoprotein extracted from cells of Saccharomyces cerevisiae, which is the traditional source. Consequently K. marxianus cultivated on a low-cost lactose-based medium such as whey, a lactose-rich clean waste of the dairy industry, could be developed as a source of bioemulsifier for use in the food industry.     

Application of centrifugal force to the extraction and separation of parasorboside and gerberin from Gerbera hybrida

A method for the separation of parasorboside and gerberin from the ornamental plant Gerbera hybrida (Asteraceae) has been developed. The two closely related glucosides were extracted using an Extrachrom instrument, a prototype multi-functional separation tool equipped with an extraction chamber. The rotation planar extraction procedure was compared with that of a medium pressure solid-liquid extraction system. The resulting extracts were pre-purified using rotation planar chromatography and the results compared with those obtained using medium pressure liquid chromatography with silica gel as the stationary phase and a mobile phase of methanol:ethyl acetate:tetrahydrofuran at selectivity point Ps = 111 with 1% formic acid as modifier. The title compounds were isolated from the purified extracts by TLC and their structures confirmed by 1H- and 13C-NMR spectroscopy.     

Photosensitizing effect of protoporphyrin IX in pigmented melanoma of mice

No fluorescence of protoporphyrin IX (PpIX) was measured using a fiber optic probe in pigmented B16F10 melanoma in mice after topical application of 5-aminolevulinic acid methylester (ALA-Me). However, chemical extraction of tissues excised from mice after intratumoral administration of ALA-Me or its parent compound ALA revealed that this tumor had the capability to produce PpIX. Small amounts of endogenous porphyrins, mainly PpIX, were found in the melanoma not treated with these drugs. Topical application of ALA-Me followed by exposure with laser light (633nm) delayed the growth of the tumors slightly. Light alone also had a significant effect on the tumor growth.     

植物细胞培养—分离耦合过程研究进展

本文评述了近10年来植物细胞培养-分离耦合过程的研究及应用,就如何深入开展植物细胞培养-分离耦合过程的研究提出了几点建议。     

用超临界流体技术萃取分离香茅油的研究

作者用超临界流体循环萃取装置对香茅油在超临界CO_2中的溶解度以及香茅油中主要成份香茅醛、香茅醇和香叶醇的分离特性进行研究。分别得出溶解度、分离特性与萃取压力、温度的关系及其相应的数学关联式。用这些关联式计算的结果符合工程需要,为单离香茅油工业化生产提供了理论基础。     

一种快速握取植物总DNA的良好材料与方法

本文介绍以种子植物幼嫩胚乳为材料,用改良的氯仿－异戊醇快速提取方法提取植物总ＤＮＡ。这种材料和方法,细胞磨碎容易,ＤＮＡ收取量大,操作简便,适用于遗传操作及教学实验等方面的ＤＮＡ快速制备。     

Quantitating adenylate nucleotides in diverse organisms

Quantitation of cellular adenylate levels (i.e., ATP, ADP, AMP) has widespread applications in physiological, metabolic and energetic studies. We have compared classical adenylate extraction procedures (i.e., perchloric acid, boiling) with a previously unreported proteinase K-based extraction technique. Our results suggest that all three techniques are comparable in soft animal tissue, but proteinase K-based extractions consistently generated higher adenylate yields from a broad range of organisms, particularly those containing a cell wall (e.g., alga, bacteria, fungi, plant).     

优美红游动菌类胡萝卜素的提取条件研究

研究了优美红游动菌(Rhodoplanes elegans)的生长以及类胡萝卜素的产生,通过比较4种广泛采用的细胞破碎方法,对优美红游动菌(Rhodoplanes elegans)类胡萝卜素的提取条件进行了研究,结果表明:类胡萝卜素的产生与菌种的生长曲线相符合,菌种活化后培养45h,类胡萝卜素的含量趋于稳定;取此时培养液,采用超声波法破碎细胞(破碎功率640W,时间10min),类胡萝卜素提取效果最好,初提液类胡萝卜素的提取率约为7.62mg·(g干菌体)^-1;酸热法对色素的破坏严重。这为进一步开发利用天然色素资源提供了一定的理论依据。     

A unique laboratory method for evaluating agro-ecosystem effects of an industrial waste product

Summary This paper describes the design, extraction, and maintenance of intact soil-core microcosms which accurately predicted biomass reduction, nutrient loss and trace element enrichment in field-grown crops amended with acidic precipitator fly ash. These agricultural microcosms sustain ecosystem-level interactions while offering a cost-effective, manageable, experimental unit which can be subjected to laboratory controls.     

Comparison of five commercial extraction kits for subsequent membrane protein profiling

Membrane proteins account for 70–80% of all pharmaceutical targets emphasizing their clinical relevance. Identification of new, differentially expressed membrane proteins reflecting distinct disease properties is thus of high importance. Unfortunately, isolation and analysis of membrane-bound proteins is hampered by their relative low abundance in total cell lysates, their frequently large size and their hydrophobic properties. We thus aimed to identify protocols that allow for highly efficient isolation and purification of membrane-bound proteins for subsequent protein profiling. We present a comparative study of different membrane protein extraction methods that vary in total protein yield between 0.02 and 4.8 mg using constant cell pellets of the colorectal carcinoma cell line SW620. We also demonstrate by means of polyacrylamide gel electrophoresis (SDS–PAGE) and Western blot analysis that the majority of commercial membrane extraction kits harbor a substantial cytosolic contamination of their membranous fraction. Based on purity of membranous fraction, protein yield, time and costs, we show superiority of two commercial extraction kits for downstream proteome analyses of membrane proteins.