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Die Hu,Jianfang Li,Qin Wu,Jia Zang,Jianqing Cheng,Minchen Wu 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.6
To improve the temperature characteristics of a mesophilic glycoside hydrolase family (GHF) 11 xylanase AoXyn11A from Aspergillus oryzae, both introduction of a disulfide bridge and the substitution of a specific amino acid were carried out by in silico design and site-directed mutagenesis. Based on the analysis of a known crystal structure of thermophilic xylanase TlXynA from Thermomyces lanuginosus, and the alignment of primary structures between AoXyn11A and TlXynA, one mutant AoXyn11AM with a disulfide bridge (Cys108–Cys152) was designed by replacing the Ser108 and Asn152 of AoXyn11A with Cys residues, respectively. Additionally, based on the analysis of amino acid B-factor values, another mutant AoXyn11AM-G22A was predicted by substituting Gly22 of AoXyn11AM (having the maximum B-factor value of 69.25 Å, with the corresponding Ala23 of TlXynA. Thereafter, two mutant xylanase-encoding genes, Aoxyn11AM and Aoxyn11AM-G22A, were constructed by site-directed mutagenesis. Aoxyn11A and two mutant genes were expressed in E. coli BL21(DE3) respectively, and three expressed recombinant xylanases, reAoXyn11A, reAoXyn11AM and reAoXyn11AM-G22A, were purified to homogeneity. The temperature optima of reAoXyn11AM and reAoXyn11AM-G22A were 60 and 65°C, respectively, being 5 and 10°C higher than that of reAoXyn11A. Their thermal inactivation half-lives at 50°C were 1.8- and 8.4-folds longer than that of reAoXyn11A. There were no obvious alterations after mutations in specific activity and enzymatic properties, except for the temperature characteristics.
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The Design and Implementation of HSP Real-time Prediction Technology Suitable for TBM Construction
( Song Lu ),( Cangsong Li ),( Xu Wang ),( Jianfang Ding ) 대한지질공학회 2019 대한지질공학회 학술발표회논문집 Vol.2019 No.2
In order to realize the real-time geological prediction that suitable for TBM construction tunnel, this paper take HSP geological prediction technology which can directly use rock-breaking vibrations generated by TBM cutters as foundation, and optimized the hardware and software of HSP system by analyzing the TBM fuselage and tunneling characteristics, so as to make it miniaturized, automated and intellectualized. The system has been installed on TBM for intelligent control and real-time prediction. During the TBM tunneling process: firstly, seismic reflection signals are collected continuously or frequently by geophones located in tunnel contours; secondly, the reflection characteristic parameters of stratum ahead of the tunnel face are acquired in real time through automatic processing; next, the effective pick-up and identification of unfavorable geological reflection interfaces are completed by intelligent recognition technology, which finally realized the prediction of unfavorable geological conditions. Therefore, through the optimization design of HSP method that suitable for TBM tunnel construction, real-time detection of the location, scale and properties of the unfavorable geological body can be achieved. This proposed technology can plays a certain role in promoting the efficient construction of TBM, which is of great significance.
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