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The chemical similarity of cellulose and chitin supports the idea that their corresponding hydrolytic enzymes would bind β-1,4-linked glucose residues in a similar manner. A structural and mutational analysis was performed for the plant cellulolytic enzyme BGlu1 from Oryza sativa and the insect chitinolytic enzyme OfHex1 from Ostrinia furnacalis. Although BGlu1 shows little amino-acid sequence or topological similarity with OfHex1, three residues (Trp490, Glu328, Val327 in OfHex1, and Trp358, Tyr131 and Ile179 in BGlu1) were identified as being conserved in the +1 sugar binding site. OfHex1 Glu328 together with Trp490 was confirmed to be necessary for substrate binding. The mutant E328A exhibited a 8-fold increment in K m for (GlcNAc)2 and a 42-fold increment in K i for TMG-chitotriomycin. A crystal structure of E328A in complex with TMG-chitotriomycin was resolved at 2.5 Å, revealing the obvious conformational changes of the catalytic residues (Glu368 and Asp367) and the absence of the hydrogen bond between E328A and the C3-OH of the +1 sugar. V327G exhibited the same activity as the wild-type, but acquired the ability to efficiently hydrolyse β-1,2-linked GlcNAc in contrast to the wild-type. Thus, Glu328 and Val327 were identified as important for substrate-binding and as glycosidic-bond determinants. A structure-based sequence alignment confirmed the spatial conservation of these three residues in most plant cellulolytic, insect and bacterial chitinolytic enzymes.

To obtain both high ionization and high deposition rate, a modified global model for a continuous high-power DC magnetron sputtering (C-HPMS) is established by considering the continuous generation of the hot electrons and the high temperature caused by continuous high-power discharge. The results show that the plasma density is on the order of 1019 m−3 for power densities of only 183 W cm−2 (Al) and 117 W cm−2 (Cu). The ionization rate exceeds 90% of high-power impulse magnetron sputtering (HiPIMS) (peak power density of 564 W cm−2) for a DC power density of 180 W cm−2, and the total diffusion fluxes of the two targets are 26 (Al) and 30 (Cu) times that of conventional HiPIMS, leading to very high deposition rates. The work provides a theoretical basis for the realization of C-HPMS and gives an enlightenment to the development of deposition equipment for continuous high-power discharges.

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