Optimization of alkaline protease production from Shewanella oneidensis MR‐1 by response surface methodology

BACKGROUND: The proteases are among the most important groups of enzymes. Therefore, it is important to produce inexpensive and optimized media for large‐scale commercial production. In the present work, three different Shewanella species were screened on skim milk agar medium for their ability to produce alkaline protease. The effects of different culture conditions were optimized for alkaline protease production by S. oneidensis MR‐1 using a Box–Behnken design combined with response surface methodology (RSM). RESULTS: Highest yield (112.90 U mL^?1) of protease production was obtained at pH 9.0, a temperature of 30 °C, glucose (12.5 g L^?1), tryptone (12.5 g L^?1) and an incubation period of 36 h. A second‐order polynomial regression model was used for analysis of the experiment. The experimental values were in good agreement with predicted values, with correlation coefficient 0.9996. CONCLUSION: Carbon and nitrogen, pH, temperature and incubation period were chosen as the main factors to be used in an experimental design for optimization to produce low‐cost enzymes, potentially for use on an industrial scale. A 60% increase in enzyme activity was achieved in the optimized medium compared with the original medium. Copyright © 2008 Society of Chemical Industry     

A Multi-service Group Key Management Scheme for Stateless Receivers in Wireless Mesh Networks

Wireless mesh networks facilitate the development of the many group oriented applications by extending the coverage area of the group communication. Group communication in a wireless mesh network is complicated due to dynamic intermediate mesh points, access control for communications between different administrative domains, and the absence of a centralized network controller. In this study, we propose a topology-matching decentralized multi-service group key management scheme for wireless mesh networks. It allows service providers to update and deliver their group keys to valid members in a distributed manner using the identity-based encryption scheme. The analysis result indicates that the proposed scheme has advantages with regard to the rekeying cost and storage overhead for a member and a mesh point in multi-sender group communication environments. The stateless property is also achieved such that a stateless member, who could not be constantly online, can easily decrypt the rekeying messages without recording the past history of transmission.     

Spectroscopic and Theoretical Study on Electronically Modified Chromophores in LOV Domains: 8‐Bromo‐ and 8‐Trifluoromethyl‐Substituted Flavins

Two chemically synthesized flavin derivatives, 8‐trifluoromethyl‐ and 8‐bromoriboflavin (8‐CF₃RF and 8‐BrRF), were photochemically characterized in H₂O and studied spectroscopically after incorporation into the LOV domain of the blue light photoreceptor YtvA from Bacillus subtilis. The spectroscopic studies were paralleled by high‐level quantum chemical calculations. In solution, 8‐BrRF showed a remarkably high triplet quantum yield (0.97, parent compound riboflavin, RF: 0.6) and a small fluorescence quantum yield (0.07, RF: 0.27). For 8‐CF₃RF, the triplet yield was 0.12, and the fluorescence quantum yield was 0.7. The high triplet yield of 8‐BrRF is due to the bromine heavy atom effect causing a stronger spin–orbit coupling. Theoretical calculations reveal that the decreased triplet yield of 8‐CF₃RF is due to a smaller charge transfer and a less favorable energetic position of T₂, required for intersystem crossing from S₁ to T₁, as an effect of the electron‐withdrawing CF₃ group. The reconstitution of the LOV domain with the new flavins resulted in the typical LOV photochemistry, consisting of triplet state formation and covalent binding of the chromophore, followed by a thermal recovery of the parent state, albeit with different kinetics and photophysical properties.     

A study on the development of a substitution process by powder metallurgy in automobile parts

Powder metallurgy processes using sintering are able to form net-shaped products and have been used widely in the production of automobile parts to improve productivity. However, the toughness of powder products is generally poor because they contain pores. Therefore, forged products are used in parts subjected to severe fatigue loads, but in the case of powder products having high toughness, they could substitute for forged products. In this study, the choice of powder materials and production processes, including mixing, compaction, sintering, and heat treatment are studied to produce the clutch disc spline hub of an automobile. For this, three types of materials are selected and processed and the mechanical properties and microstructure of the sample are investigated, along with the performance of a dynamic test carried out under real conditions.     

Predicting metamorphic relations for testing scientific software: a machine learning approach using graph kernels

Comprehensive, automated software testing requires an oracle to check whether the output produced by a test case matches the expected behaviour of the programme. But the challenges in creating suitable oracles limit the ability to perform automated testing in some programmes, and especially in scientific software. Metamorphic testing is a method for automating the testing process for programmes without test oracles. This technique operates by checking whether the programme behaves according to properties called metamorphic relations. A metamorphic relation describes the change in output when the input is changed in a prescribed way. Unfortunately, finding the metamorphic relations satisfied by a programme or function remains a labour‐intensive task, which is generally performed by a domain expert or a programmer. In this work, we propose a machine learning approach for predicting metamorphic relations that uses a graph‐based representation of a programme to represent control flow and data dependency information. In earlier work, we found that simple features derived from such graphs provide good performance. An analysis of the features used in this earlier work led us to explore the effectiveness of several representations of those graphs using the machine learning framework of graph kernels, which provide various ways of measuring similarity between graphs. Our results show that a graph kernel that evaluates the contribution of all paths in the graph has the best accuracy and that control flow information is more useful than data dependency information. The data used in this study are available for download at http://www.cs.colostate.edu/saxs/MRpred/functions.tar.gz to help researchers in further development of metamorphic relation prediction methods. Copyright © 2015 John Wiley & Sons, Ltd.     

A preliminary study on the development of an easy method for beef freshness using a cyclic voltammetric system

This study was carried out to develop an easy method for beef freshness using a cyclic voltammetric system. Prime graded (USDA) beef loin at 48 h postmortem was cut into pieces 3 cm thick, packaged into commercial plastic zipper bag, and then stored at 4 °C for 0, 3, 6 and 9 days. A commercial cyclic voltammetry and data processing software were used for this study and the interdigitated electrode was fabricated according to semiconductor manufacturing. The homogenated beef was filtered through Whatman No. 1 filter paper and 250 μL of the filtrate was applied on the interdigitated electrode sensor chip for electrochemical detection. Electrochemical signals changed as storage periods increased. The shape of the cyclic voltammogram changed from a roundish curve to a shapely curve. Redox values (V) also increased during storage. Electrochemical signals matched well with lipid oxidation values (Thiobarbituric acid reactive substances: TBARS). A linear relationship between redox value and TBARS was obtained with a correlation factor of 0.957. Thus, the electrochemical signal of a cyclic voltammetric system can be used as an indicator of beef freshness.     

Relationship of Carcass Weight to Muscle Fiber Characteristics and Pork Quality of Crossbred (Korean Native Black Pig × Landrace) F2 Pigs

A total of 168 carcasses of crossbred (Korean native black pig × Landrace) F₂ pigs were divided into four carcass weight groups (A, 70–79 kg; B, 80–89 kg; C, 90–99 kg; D, 100–109 kg). The influence of carcass weight on muscle fiber characteristics and meat quality traits of pork, from the longissimus dorsi, were investigated. Carcass traits such as loin-eye area and back-fat thickness were found to increase with increasing carcass weight. Fat content, sarcomere length, drip loss, and meat color values (L*, a*, and b*) also increased with increasing carcass weight. However, moisture content and Warner–Bratzler shear force decreased with increasing carcass weight. A decrease in the compositions of type I and IIA fibers and an increase in the composition of type IIB fibers were observed with increasing carcass weight. The heavier carcasses showed type I and IIB fibers of bigger size and a smaller size of type IIA fibers. Fiber number percentage and the diameter of type IIB showed a significant positive correlation with carcass weight, fat content, and lightness (p?<?0.05). Fiber diameter of type IIB was positively correlated with loin-eye area, but was negatively correlated with moisture content. However, muscle fiber characteristics had no correlation with back-fat thickness, ultimate pH, Warner–Bratzler shear force, and b* value. In conclusion, crossbred pig which has of high composition and size of type IIB has high lightness (L* value) and water-holding capacity (drip loss). Therefore, excessively high weight of carcass has influence on pale and exudative properties in pig because of muscle fiber characteristics, especially type IIB fiber.