Isolation,cloning, and characterization of the 2S albumin: A new allergen from hazelnut

Scope: 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity. Methods: 2S albumin from hazelnut extract was purified using size exclusion chromatography and RP‐HPLC. After N‐terminal sequencing, degenerated and poly‐d(T) primers were used to clone the 2S albumin sequence from hazelnut cDNA. After expression in Escherichia coli and affinity purification, IgE reactivity was evaluated by Immunoblot/ImmunoCAP (inhibition) analyses using sera of nut‐allergic patients. Results: N‐terminal sequencing of a ～10 kDa peak from size exclusion chromatography/RP‐HPLC gave two sequences highly homologous to pecan 2S albumin, an 11 amino acid (aa) N‐terminal and a 10aa internal peptide. The obtained clone (441 bp) encoded a 147aa hazelnut 2S albumin consisting of a putative signal peptide (22 aa), a linker peptide (20 aa), and the mature protein sequence (105 aa). The latter was successfully expressed in E. coli. Both recombinant and natural 2S albumin demonstrated similar IgE reactivity in Immunoblot/ImmunoCAP (inhibition) analyses. Conclusion: We confirmed the postulated role of hazelnut 2S albumin as an allergen. The availability of recombinant molecules will allow establishing the importance of hazelnut 2S albumin for hazelnut allergy.     

The effect of multiple yeasts co-inoculations on Sauvignon Blanc wine aroma composition,sensory properties and consumer preference

Consumer acceptance testing has been only recently applied in wine research, to assess wine sensory attributes that affect hedonic liking. The aim of this study was to investigate the effect of Saccharomyces yeast co-inoculations on wine volatile composition and sensory profiles, and to determine if differences were sufficient enough to affect consumer acceptance. Fermentations were conducted using two- and three-yeast co-inoculations, and single strains. Yeast inocula differed substantially in volatile thiols and other flavour compounds, and in their sensory properties. Wines from four yeast inocula which showed large sensory differences were subjected to consumer testing by 120 consumers, with differences in overall liking found. Four clusters of consumers were identified, with one group strongly preferring the two-yeast co-inoculated wine with an intermediate sensory profile, while another group favoured the wine made using the three-yeast co-inoculation. This study has demonstrated that the yeast inoculum used to conduct fermentation affects consumer acceptance.     

CLONING OF A YEAST GENE WHICH CAUSES PHENOLIC OFF-FLAVOURS IN BEER

A gene (POF1) has been cloned, which confers upon yeast (Saccharomyces cerevisiae) the ability to decarboxylate phenolic acids such as ferulic and trans-cinnamic acid. This property was previously shown to be a cause of phenolic off-flavour production in wort fermentations. The identity of the cloned gene was confirmed as POF1 by gene disruption techniques. Southern blotting of total genomic DNA revealed that sequences homologous to POF1 are conserved in Pof^? brewing strains of Sacch. cerevisiae. The transformation of a Pof^? lager strain with the cloned POF1 gene led to the production of an aroma characteristic of a phenolic off-flavour, when the transformed strain was used in wort fermentations. This latter observation suggests that the Pof^? phenotype of brewers' yeast is specifically due to the absence of a functional POF1 gene.     

甘蔗糖蜜生产核苷酸的分离方法研究

本文介绍了利用离子交换层析技术分离甘蔗糖蜜生产的核苷酸之现实性与可行性。探讨了分离核苷酸的最佳工艺条件。     

Family M42 aminopeptidase from the syntrophic bacterium Symbiobacterium thermophilum: characterization using recombinant protein

The chromosomal DNA of the syntrophic thermophile Symbiobacterium thermophilum contains open reading frames of the genes encoding family M42 aminopeptidases, Pep1079, Pep1080, and Pep1081. To characterize these peptidases, the genes were cloned into Escherichia coli and overexpressed. Our experiments using the recombinant proteins confirmed that Pep1079, Pep1080, and Pep1081 are components of arginyl or lysinyl aminopeptidases that require Co2+ for enzymatic activity. Coexistence of Pep1079 and Pep1080 is necessary for expressing high peptidase activity. Pep1081 enhances the activity of Pep1079 and Pep1080.     

Effect of post-induction substrate oscillation on recombinant alkaline phosphatase production expressed in Escherichia coli

Microorganisms are exposed to fast changes in microenvironment in large scale bioreactors. Because of their fast response to the changes, overall performance of biological system in small scale differs from large scale. Hence the variations in the environment that microorganisms are living in are mimicked in small scale. For this purpose one way is to feed substrate into the bioreactor in an oscillatory profile. In this work two different types of triangular oscillatory feeding profiles were applied as the post induction feeding strategy in intracellular recombinant alkaline phosphatase production expressed in Escherichia coli to find out if this biological system behaves in inhomogeneous environment differently. On line and offline measurements provide evaluation of product quality and quantity. Then the results of the experiments were compared with those of the control run at which constant feeding rate was executed. The results showed that oscillatory feeding at which cells were not starved led to higher yield of protein per substrate (0.027 C-mol/C-mol) and higher activity per protein (0.79 U/mg) when compared to a constant feeding rate (0.011 C-mol/C-mol and 0.11 U/mg).     

酵母生物制剂对果蔬采后病害形成的影响

近年来的研究表明,酵母菌株的生物制剂对果蔬采后病害具有良好的抑制作用,如用其替代杀菌剂可减少农药使用量,降低产品的农药残留,保护生态环境,保证食品安全。本文介绍了酵母对病原微生物的拮抗作用机制,并对影响拮抗剂使用效果的一些重要因素,以及对如何提高拮抗效果的措施进行了深入的讨论。该领域具有很好的研究和应用前景。     

鲜奶蛋糕卫生标准的研究

对４４份市售鲜奶蛋糕样品的卫生指标进行了检测,结合鲜奶蛋糕的生产销售及检测结果,对其酸价、过氧化值、菌落总数及大肠菌群等卫生指标提出了建议值,并建议在ＧＢ７０９９中增加酵母菌数的卫生指标,从而为鲜奶蛋糕的监督管理提供法律依据。     

New type of pressurized cultivation method providing oxygen for piezotolerant yeast

For efficient oxygen supply to pressurized culture, we developed a method using a highly pressurized membrane reactor with an air-saturated medium circulation system. The new method increased the cell growth of aerobic yeast approximately 20 folds larger than that in the case of using a conventional method.