高压电场对液化酶的活性影响及其机理探讨

研究在高压电场下不同的电场强度、温度、处理时间对液化酶活性影响。25℃时以22kV／cm的场强处理30s，酶活降低60％；21℃时以15kV／cm处理1min，酶活降低30．8％，处理10min，酶活降低45.17％；以15kV／cm的场强处理样品30s，21℃时酶活降低23．55％；75℃时酶活降低77．05％，并对各现象机理进行了探讨。     

Construction of covalently attached enzyme multilayer films based on the photoreaction of diazo-resins and glucose oxidase

A novel and facile approach to construct multilayered glucose oxidase (GOx) films on the surface of quartz or CaF₂ slides as well as gold electrodes for use as biosensing interfaces is described. Diazo-resins (DAR) as polycation and glucose oxidase as polyanion were alternately deposited into a multilayer structure using layer-by-layer self-assembly technique based on electrostatic interaction as driving force. Upon near UV irradiation, the adjacent interfaces of the multilayer reacted to form a crosslinking structure which greatly improved the stability of the enzyme films. These changes was monitored and confirmed by UV-vis and IR spectroscopy. Ellipsometric measurements reveal that the enzymes formed sub-molecule layers, and the thickness of the film shows a linear relationship with the number of assembled layers, demonstrating a spatially well-ordered manner in multilayer structure. The covalently attached enzyme multilayer film has a highly permeable structure, and can be used as biosensing interface. Electrochemical and analytical behavior of the enzyme electrodes was studied by cyclic voltammetry (CV) in the presence or absence of glucose. The sensitivity of the enzyme-modified electrodes was estimated through the analysis of voltammetric signals, which can be fine turned to the desired level by adjusting the number of attached bilayers.     

Synthesis of l-theanine using enzyme/mesoporous silica conjugates under high pH conditions

We present a new enzymatic process for synthesis of l-theanine using glutaminase combined with immobilization technique on a mesoporous silica (MPS). The MPS was firstly attempted to modify with zirconia in order to enhance the durability against the reaction under high pH conditions. The glutaminase on the MPS successfully catalyzed the reaction for the synthesis of l-theanine. The glutaminase/MPS conjugate was subsequently recovered and employed for the reaction again. The conjugate showed the corresponding activity to the first synthesis. This indicates that the conjugate functions as a catalyst for synthesis of l-theanine, having the operational stability sufficient for reuse.     

Biodegradation of anthracene by Aspergillus fumigatus

An anthracene-degrading strain, identified as Aspergillus fumigatus, showed a favorable ability in degradation of anthracene. The degradation efficiency could be maintained at about 60% after 5d with initial pH of the medium kept between 5 and 7.5, and the optimal temperature of 30 °C. The activity of this strain was not affected significantly by high salinity. Exploration on co-metabolism showed that the highest degradation efficiency was reached at equal concentration of lactose and anthracene. Excessive carbon source would actually hamper the degradation efficiency. Meanwhile, the strain could utilize some aromatic hydrocarbons such as benzene, toluene, phenol etc. as sole source of carbon and energy, indicating its degradation diversity. Experiments on enzymatic degradation indicated that extracellular enzymes secreted by A. fumigatus could metabolize anthracene effectively, in which the lignin peroxidase may be the most important constituent. Analysis of ion chromatography showed that the release of anions of A. fumigatus was not affected by addition of anthracene. GC-MS analysis revealed that the molecular structure of anthracene changed with the action of the microbe, generating a series of intermediate compounds such as phthalic anhydride, anthrone and anthraquinone by ring-cleavage reactions.     

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.     

Uniform culture in solid-state fermentation with fungi and its efficient enzyme production

Solid-state fermentation (SSF) has attracted a lot of interest for carrying out high-level protein production in filamentous fungi. However, it has problems such as the fermentation heat generated during the culture in addition to the reduced mobility of substances. These conditions lead to a nonuniform state in the culture substrate and result in low reproducibility. We constructed a non-airflow box (NAB) with a moisture permeable fluoropolymer membrane, thereby making it possible to control and maintain uniform and optimal conditions in the substrate. For the NAB culture in Aspergillus oryzae, temperature and water content on/in the whole substrate were more consistent than for a traditional tray box (TB) culture. Total weight after the culture remained constant and dry conditions could be achieved during the culture. These data demonstrate the possibility of growing a uniform culture of the whole substrate for SSF. The NAB is advantageous because it allows for the control of exact temperature and water content in the substrate during the culture by allowing vapor with latent heat to dissipate out of the box. In addition, several enzymes in the NAB culture exhibited higher production levels than in the TB culture. We believe that culturing in the constructed NAB could become a standard technique for commercial SSF.     

Nucleotides and their degradation products during processing of dry-cured ham, measured by HPLC and an enzyme sensor

The aim of this work was to study how nucleotide degradation during the processing of dry-cured ham is affected when using three types of salting (100% NaCl; 50% NaCl and 50% KCl; 55% NaCl, 25% KCl, 15% CaCl₂ and 5% MgCl₂). Divalent salts in the salting mixture depressed the breakdown rate from the beginning of the process (salting and post-salting) up to the ripening stage (7 months) when the inosine (Ino), hypoxanthine (Hx) and xanthine (X) concentrations matched for the three treatments. The evolution of Hx and Hx + X were analysed by HPLC and an enzyme sensor, respectively, during processing. Time and temperature conditions during the curing time did not affect Hx stability. The usefulness of the enzyme sensor was confirmed and it is a practical tool to determine Hx + X in dry-cured ham, as an index of minimum curing time. A good correlation between enzyme sensor and HPLC data was observed.     

自动控制脉冲强光杀菌装置的研制

采用单片机控制技术，研制了自动控制脉冲强度杀菌装置。每个光脉冲的输入能量可达６００～２２００Ｊ；光脉冲宽度经测定小于８００μＳ。３０个光脉冲能使大肠杆菌减少５个数量级，４０个光脉冲以使蛋白酶的活力钝化９０％，脉冲强光杀菌可事负透明塑料薄膜包装的切片面包的保存期延长１倍以上，脉冲强光杀菌是一种很有发展前途的冷杀菌技术。     

酶制剂在食品中的应用概况

本文综述了酶制剂在加工食品原料、改善品质、提高附加值、保鲜、检测、解毒等方面的应用概况 ,并展望其未来。