Gene Cloning, Heterologous Expression and Biochemical Characterization of A Novel Extracellular Lipase from Rhizopus Oryzae KU45.

BACKGROUND: Lipases secreted from various Rhizopus oryzae strains were previously expressed in Escherichia coli, Pichia pastoris, and Saccharomyces cerevisiae and was shown to have distinct activities in response to different temperatures, metal ions, organic solvents, and specific substrates. However, until now, no other research biochemically characterized the functions of extracellular pro-lipase in a novel Rhizopus oryzae KU45. OBJECTIVES: Characterization of a novel extracellular lipase from fungus R. oryzae KU45 after heterologous expression in E. coli BL21 (DE3) strain. MATERIALS AND METHODS: An extracellular lipase producing fungus was isolated from a soil sample and identified as a strain of R. oryzae by partial 18S rRNA gene sequencing. It was named as R. oryzae KU45. The lipase gene of KU45 was cloned into pET-28a expression vector and expressed in E. coli as inclusion bodies. The recombinant lipase was purified, refolded and characterized. RESULTS: The lipase exhibited maximum activity at 45ºC, at slightly alkaline pH. It showed a broad substrate specificity acting on p-nitrophenyl esters with C8-C16 acyl groups as substrates and, many of the organic solvents and metal ions tested did not have any adverse effects on the enzyme activity. CONCLUSIONS: High stability, broad substrate specificity and activity at mesophilic temperatures in the presence of organic solvents, and metal ions make the extracellular lipase of KU45 a candidate for various biotechnological applications.

Humidity effects on anisotropic nanofriction behaviors of aligned carbon nanotube carpets.

We evaluated the interfacial properties of vertically and transversely aligned multiwalled carbon nanotubes (CNTs) carpets using atomic force microscopy (AFM) under ambient (26%-52% relative humidity (RH)), mild (10% RH), and dry conditions (<3.3% RH). The frictional forces on a transversely aligned CNT (TAMWCNT) surface are lower than those on a vertically aligned CNT (VAMWCNT) surface, and they decrease as the relative humidity decreases for both TAMWCNT and VAMWCNT surfaces. Similar trends are found for adhesion forces on both surfaces. Contact mechanics theories are applied and compared in an attempt to better understand these results. The tribological behavior of CNT carpets unveiled in this paper is expected to inspire tunable friction surface engineering strategies.

Continuous carbon nanotube reinforced composites.

Carbon nanotubes are considered short fibers, and polymer composites with nanotube fillers are always analogues of random, short fiber composites. The real structural carbon fiber composites, on the other hand, always contain carbon fiber reinforcements where fibers run continuously through the composite matrix. With the recent optimization in aligned nanotube growth, samples of nanotubes in macroscopic lengths have become available, and this allows the creation of composites that are similar to the continuous fiber composites with individual nanotubes running continuously through the composite body. This allows the proper utilization of the extreme high modulus and strength predicted for nanotubes in structural composites. Here, we fabricate such continuous nanotube polymer composites with continuous nanotube reinforcements and report that under compressive loadings, the nanotube composites can generate more than an order of magnitude improvement in the longitudinal modulus (up to 3,300%) as well as damping capability (up to 2,100%). It is also observed that composites with a random distribution of nanotubes of same length and similar filler fraction provide three times less effective reinforcement in composites.

Time and temperature dependence of multi-walled carbon nanotube growth on Inconel 600.

The growth kinetics of vertically aligned multi-walled carbon nanotubes (MWNTs) on conductive substrates is investigated by synthesizing MWNTs at different growth temperatures and measuring ex situ the length of the nanotubes as a function of growth duration. A typical 'root growth' mechanism (saturation of MWNT lengths with increasing growth duration) was observed. The value of the activation energy (E(a)≈136 ± 5 kJ mol(-1)) of the MWNT growth reaction is suggestive of carbon diffusion through the bulk of the iron catalyst particle versus diffusion on its surface. These findings will help in optimizing MWNT growth on conductive substrates for various applications.

Fatigue resistance of aligned carbon nanotube arrays under cyclic compression.

Structural components subject to cyclic stress can succumb to fatigue, causing them to fail at stress levels much lower than if they were under static mechanical loading. However, despite extensive research into the mechanical properties of carbon nanotube structures for more than a decade, data on the fatigue behaviour of such devices have never been reported. We show that under repeated high compressive strains, long, vertically aligned multiwalled nanotubes exhibit viscoelastic behaviour similar to that observed in soft-tissue membranes. Under compressive cyclic loading, the mechanical response of the nanotube arrays shows preconditioning, characteristic viscoelasticity-induced hysteresis, nonlinear elasticity and stress relaxation, and large deformations. Furthermore, no fatigue failure is observed at high strain amplitudes up to half a million cycles. This combination of soft-tissue-like behaviour and outstanding fatigue resistance suggests that properly engineered nanotube structures could mimic artificial tissues, and that their good electrical conductivity could lead to their use as compliant electrical contacts in a variety of applications.

Direct growth of aligned carbon nanotubes on bulk metals.

There are several advantages of growing carbon nanotubes (CNTs) directly on bulk metals, for example in the formation of robust CNT-metal contacts during growth. Usually, aligned CNTs are grown either by using thin catalyst layers predeposited on substrates or through vapour-phase catalyst delivery. The latter method, although flexible, is unsuitable for growing CNTs directly on metallic substrates. Here we report on the growth of aligned multiwalled CNTs on a metallic alloy, Inconel 600 (Inconel), using vapour-phase catalyst delivery. The CNTs are well anchored to the substrate and show excellent electrical contact with it. These CNT-metal structures were then used to fabricate double-layer capacitors and field-emitter devices, which demonstrated improved performance over previously designed CNT structures. Inconel coatings can also be used to grow CNTs on other metallic substrates. This finding overcomes the substrate limitation for nanotube growth which should assist the development of future CNT-related technologies.

Changes in forest biomass carbon storage in China between 1949 and 1998.

The location and mechanisms responsible for the carbon sink in northern mid-latitude lands are uncertain. Here, we used an improved estimation method of forest biomass and a 50-year national forest resource inventory in China to estimate changes in the storage of living biomass between 1949 and 1998. Our results suggest that Chinese forests released about 0.68 petagram of carbon between 1949 and 1980, for an annual emission rate of 0.022 petagram of carbon. Carbon storage increased significantly after the late 1970s from 4.38 to 4.75 petagram of carbon by 1998, for a mean accumulation rate of 0.021 petagram of carbon per year, mainly due to forest expansion and regrowth. Since the mid-1970s, planted forests (afforestation and reforestation) have sequestered 0.45 petagram of carbon, and their average carbon density increased from 15.3 to 31.1 megagrams per hectare, while natural forests have lost an additional 0.14 petagram of carbon, suggesting that carbon sequestration through forest management practices addressed in the Kyoto Protocol could help offset industrial carbon dioxide emissions.

[Comparison analysis between potential and actual pattern of artificial oases in arid region].

Based on theoretical analysis and demonstration research, the conception of potential pattern in the agriculture landscape of artificial oases in Xinjiang arid region and its analysis unit were discussed. The potential landscape pattern was defined as the one composed by spatial units with basic characteristics and properties which had no change or less change with the time. In agriculture landscape, soil was found to be a relatively stable element, and hence, different soil classification unit could be used to analyze the potential landscape pattern. A case study was carried out to analyze the potential and actual pattern of the artificial cases in Shihezi reclamation area by using the indexes of diversity, evenness, aggregation, mean patch elongation, patch shape fragmentation and mean patch fractal dimension. The result showed that the landscape pattern changed orderly from the potential to actual pattern, and the potential pattern could be used as the absolute criterion for researches on pattern changes in agriculture landscape.