酶箱模具设计和改讲

通过对酶箱模具设计和改进过程的分析,提出了企业在追求低投入高产出时,必须是在确保产品质量的前提下考虑效益最大化,否则是得不偿失的。提示设计人员在搞设计时,要把握住质量和投入的平衡。     

溶菌酶测定方法的改进

简化了溶菌酶测定底物的制备过程，确定了读数范围，改进传统溶菌酶活力检测方法，确定酶活力标准曲线，此方法的活力检测限灵敏。     

Two-parameter model for evaluating effectiveness factor for immobilized enzymes with reversible Michaelis-Menten kinetics

A two-parameter mathematical model was developed to calculate the effectiveness factor for immobilized enzymes in porous spherical particles. The model was resolved for reversible Michaelis-Menten kinetics, including simple Michaelis-Menten and product competitive inhibition kinetics. Since only two dimensionless moduli are involved in the model, the effectiveness factor for the three kinetic equations considered can be estimated by using only one generalized graph.     

Nanostructures for enzyme stabilization

Recent breakthroughs in nanotechnology have made various nanostructured materials more affordable for a broader range of applications. Although we are still at the beginning of exploring the use of these materials for biocatalysis, various nanostructures have been examined as hosts for enzyme immobilization via approaches including enzyme adsorption, covalent attachment, enzyme encapsulation, and sophisticated combinations of methods. This review discusses the stabilization mechanisms behind these diverse approaches; such as confinement, pore size and volume, charge interaction, hydrophobic interaction, and multipoint attachment. In particular, we will review recently reported approaches to improve the enzyme stability in various nanostructures such as nanoparticles, nanofibers, mesoporous materials, and single enzyme nanoparticles (SENs). In the form of SENs, each enzyme molecule is surrounded with a nanometer scale network, resulting in stabilization of enzyme activity without any serious limitation for the substrate transfer from solution to the active site. SENs can be further immobilized into mesoporous silica with a large surface area, providing a hierarchical approach for stable, immobilized enzyme systems for various applications, such as bioconversion, bioremediation, and biosensors.     

Effects of Additives on the Activity and Enantioselectivity of Candida rugosa Lipase in a Biphasic Medium

The effects of Li⁺, Na⁺, K⁺, Mg²⁺ and Zn²⁺ ions on the activity and enantioselectivity of Candida rugosa lipase (CRL) were investigated in a biphasic medium composed of phosphate buffer solution (containing a metal ion within a 50–500 mM concentration range) and isooctane. The hydrolytic activities of CRL towards p‐nitrophenyl acetate were measured after incubation of the enzyme in the presence of metal ions for 24 h, and they were compared to that obtained after incubation in the absence of any metal ion. The CRL activity was stimulated by the chloride salts of Li⁺, K⁺ and Mg²⁺ for all concentrations considered and the highest enhancement was achieved by Li⁺ with a 1.24–1.75 fold increase observed. The effects of metal ions on the enantioselectivity of CRL were investigated by performing the hydrolysis of racemic Naproxen methyl ester in the same biphasic medium containing Li⁺, Na⁺, K⁺, Mg²⁺ and Zn²⁺ ions. The addition of metal ions increased the hydrolysis rate by ca. 1.31–1.45 fold relative to the control, whereas the enantiomeric excess of product increased slightly in the presence of the metal ions. The effect of Triton X‐100 on the activity and enantioselectivity of the CRL was also investigated by employing 0.05, 0.1, 0.2, 0.3, 0.4, and 0.5 mM concentrations of it in phosphate buffer solution of the biphasic medium. High concentrations of Triton X‐100 stimulated the enzyme activity up to 1.66 fold after 24 h incubation. Triton X‐100 increased the hydrolysis rate almost independently of the concentration.     

Cytochemical applications of X-ray microanalysis

X-ray microanalysis (XRMA) has been applied to a wide variety of cytochemical problems, but the most valuable applications have been to the validation of cytochemical methods (by the qualitative or quantitative analysis of reaction products), and to the simultaneous localization of more than one substance, which cannot easily be achieved by using alternative methods. The latter applications involve stoichiometric studies (the quantitative relationships between reaction products and substrates), and distribution studies. Ultrastructural cytochemistry with XRMA is limited by the need to use high-brightness electron sources. Apart from the limited availability of such sources, they may cause unacceptable damage to the specimen. Preparation methods for cytochemistry using XRMA are reviewed; in principle these do not differ from those used for other cytochemical applications, but it is important not to introduce extraneous elements (from fixative, buffer, or embedding medium) into the specimen, where the additional X-ray peaks may interfere with the analysis. Quantification in XRMA of cytochemical preparations poses special problems, because the addition of the reaction product to the specimen alters the yield of continuum X rays, used for assessing the mass of the specimen, and also dilutes endogenous elements. However, measurement of ratios between characteristic elemental peaks is a useful method in X-ray microanalytical cytochemistry, and it is concluded that one of the most important attributes of XRMA for cytochemical purposes is the ease with which the substances of interest can be measured.     

NASICON-based solid-electrolyte cell as transducer for enzyme sensors

Enzyme sensors for glucose and uric acid have been developed based on a solid-electrolyte cell using NASICON (Na₃Zr₂Si₂PO₁₂). These potentiometric devices respond reversibly to glucose and uric acid over a concentration range of two orders of magnitude with sensitivities of −54 and −52 mV/decade, respectively. The sensors can be used for a batch-type as well as a flow-through-type measuring system. Primarily the sensors respond to the H₂O₂ that is produced by the enzymatic reactions. The role of the three-phase region, electrolyte solution, sensing electrode metal and NASICON has been investigated. The liquid electrolyte/sensing electrode metal interface is found to work as a potential-determining as well as a rate-determining interface for the enzyme sensor.     

Purification and biochemical characterization of chymotrypsin from the viscera of Monterey sardine (Sardinops sagax caeruleus)

Chymotrypsin was isolated from the viscera of Monterey sardine by ammonium sulphate fractionation, gel filtration, and ionic exchange chromatography. The approximate molecular weight was 26,000 and its isoelectric point was about 5. Identity as chymotrypsin was established by its catalytic specificity for amide or ester bonds on the synthetic substrates succinyl-l-ala-ala-pro-l-pheilalanine-p-nitroanilide and benzoyl-l-tyrosine-ethyl-ester, showing esterase activity 3.2-fold higher than amidase. It was inhibited by phenylmethylsulfonyl-fluoride and soybean trypsin inhibitor, partly inhibited by the specific chymotrypsin inhibitor N-toluenesulfonyl-l-phenylalanine chloromethyl-ketone, but not inhibited by EDTA or Benzamidine. Chymotrypsin showed its maximum activity at pH 8.0 and 50 °C for the hydrolysis of SAAPNA. The Michaelis–Menten constant was 0.074 mM with a catalysis constant of 18.6 seg⁻¹, and catalytic efficiency of 252 seg⁻¹ mM⁻¹. Results indicated that Monterey sardine chymotrypsin is a good catalyst and could be used as a biotechnological tool in food processing and using sardine industry wastes as a material for production of fine reagents.     

ENZYMATIC HYDROLYSIS OF STARCH BY USING A SONIFIER

In the present study, the ultrasonication method was used to investigate the effect of ultrasonic energy on the hydrolysis of corn, rice, and wheat starch by using the alpha-amylase enzymes produced by Bacillus species and Bacillus licheniformis. The effects of sonifier operation variables such as duty cycle and acoustic power rate on the stability of alpha-amylase enzymes and hydrolysis degrees of three types of starches were investigated at a temperature of 40°C and pH 6.5. To determine the effect of temperature with sonication on the hydrolysis process, wheat starch hydrolysis experiments were also carried out at a temperature of 50°C. Then, the relation between duty cycle and enzyme stability during hydrolysis for each enzyme at 50°C was expressed by a zero-order inactivation model.