Mutational analysis to identify the residues essential for the inhibition of N-acetyl glutamate kinase of Corynebacterium glutamicum.

N-acetyl glutamate kinase (NAGK) is a key enzyme in the synthesis of L-arginine that is inhibited by its end product L-arginine in Corynebacterium glutamicum (C. glutamicum). In this study, the potential binding sites of arginine and the residues essential for its inhibition were identified by homology modeling, inhibitor docking, and site-directed mutagenesis. The allosteric inhibition of NAGK was successfully alleviated by a mutation, as determined through analysis of mutant enzymes, which were overexpressed in vivo in C. glutamicum ATCC14067. Analysis of the mutant enzymes and docking analysis demonstrated that residue W23 positions an arginine molecule, and the interaction between arginine and residues L282, L283, and T284 may play an important role in the remote inhibitory process. Based on the results of the docking analysis of the effective mutants, we propose a linkage mechanism for the remote allosteric regulation of NAGK activity, in which residue R209 may play an essential role. In this study, the structure of the arginine-binding site of C. glutamicum NAGK (CgNAGK) was successfully predicted and the roles of the relevant residues were identified, providing new insight into the allosteric regulation of CgNAGK activity and a solid platform for the future construction of an optimized L-arginine producing strain.

Experimental and numerical study of biomass flash pyrolysis in an entrained flow reactor.

Experiments on flash pyrolysis of biomass (rice husk and sawdust) were performed in an entrained flow reactor at different temperatures (700-1000 degrees C). Results show that temperature has great impact on the biomass flash pyrolysis reactions. With the increase of temperature, the gas yield increased, while the yields of char and liquid decreased. After the early stage of pyrolysis, the gas yield increased gradually which could be attributed to the increase of CO and CH(4) yields at low temperature, while to the increase of CO and H(2) yields at high temperature. A first order kinetic model was employed to describe biomass flash pyrolysis characteristics and the relevant kinetic parameters were determined. Numerical simulation of pyrolysis process was carried out, the predicted yields of produced gases agreed reasonably with the experimental data.

Experimental study on cyclone air gasification of wood powder.

In this paper, effects of the equivalence ratio (ER) and the secondary air on the gasification system were studied. The results indicate that as the ER varies in the range of 0.20-0.26, the low heating value (LHV) of the producer gas is in the range of 3.64-5.76 MJ/Nm(3), the carbon conversion is 55%-67% and the cold gas efficiency of the gasification system is 33%-47%. In contrast to the gasification without the secondary air, air staged process is a gasification method capable of increasing the LHV of the producer gas from 4.63 to 5.67 MJ/Nm(3), the carbon conversion from 65.5% to 81.2%, and the cold gas efficiency of the gasifier from 42.5% to 56.87%, while the tar content of the producer gas decreases from 13.96 to 5.6g/Nm(3). There exists an optimum ratio of the secondary air.

Characteristics of rice husk gasification in an entrained flow reactor.

Experiments were performed in an entrained flow reactor to better understand the characteristics of biomass gasification. Rice husk was used in this study. Effects of the gasification temperature (700, 800, 900 and 1000 degrees C) and the equivalence ratio in the range of 0.22-0.34 on the biomass gasification and the axial gas distribution in the reactor were studied. The results showed that reactions of CnHm were less important in the gasification process except cracking reactions which occurred at higher temperature. In the oxidization zone, reactions between char and oxygen had a more prevailing role. The optimal gasification temperature of the rice husk could be above 900 degrees C, and the optimal value of ER was 0.25. The gasification process was finished in 1.42 s when the gasification temperature was above 800 degrees C. A first order kinetic model was developed for describing rice husk air gasification characteristics and the relevant kinetic parameters were determined.