Effects of C-terminal domain truncation on enzyme properties of Serratia marcescens chitinase C.

A chitinase gene (SmChiC) and its two C-terminal truncated mutants, SmChiCG426 and SmChiCG330 of Serratia marcescens, were constructed and cloned by employing specific polymerase chain reaction (PCR) techniques. SmChiCG426 is derived from SmChiC molecule without its C-terminal chitin-binding domain (ChBD) while SmChiCG330 is truncated from SmChiC by its C-terminal deletion of both ChBD and fibronectin type III domain (FnIII). To study the role of the C-terminal domains of SmChiC on the enzyme properties, SmChiC, SmChiCG426, and SmChiCG330 were expressed in Escherichia coli by using the pET-20b(+) expression system. The His-tag affinity-purified SmChiC, SmChiCG426, and SmChiCG330 enzymes had a calculated molecular mass of 51, 46, and 36 kDa, respectively. Certain biochemical characterizations indicated that the enzymes had similar physicochemical properties, such as the optimum pH (5), temperature (37 °C), thermostability (50 °C), and identical hydrolyzing product (chitobiose) from both the soluble and insoluble chitin substrates. The overall catalytic efficiency k cat /K M was higher for both truncated enzymes toward the insoluble α-chitin, whereas the binding abilities toward the insoluble α-chitin substrate were reduced moderately. The fluorescence and circular dichroism (CD) spectroscopy data suggested that both mutants retained a similar folding conformation as that of the full-length SmChiC enzyme. However, a CD-monitored melting study showed that the SmChiCG330 had no obvious transition temperature, unlike the SmChiC and SmChiCG426.

MRI-compatible ultrasound heating system with ring-shaped phased arrays for breast tumor thermal therapy.

Therapeutic ultrasound transducers can carry out precise and efficient power deposition for tumor thermal therapy under the guidance of magnetic resonance imaging. For a better heating, organ-specific ultrasound transducers with precision location control system should be developed for tumors located at various organs. It is feasible to perform a better heating for breast tumor thermal therapy with a ring-shaped ultrasound phased-array transducer. In this study, we developed ring-shaped phased-array ultrasound transducers with 1.0 and 2.5 MHz and a precision location control system to drive the transducers to the desired location to sonicate the designated region. Both thermo-sensitive hydrogel phantom and ex vivo fresh pork were used to evaluate the heating performance of the transducers. The results showed that the ring-shaped phased array ultrasound transducers were very promising for breast tumor heating with the variation of heating patterns and without overheating the ribs.