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171.
Xuejun C. Zhang Yan Zhao Jie Heng Daohua Jiang 《Protein science : a publication of the Protein Society》2015,24(10):1560-1579
Major facilitator superfamily (MFS) is a large class of secondary active transporters widely expressed across all life kingdoms. Although a common 12‐transmembrane helix‐bundle architecture is found in most MFS crystal structures available, a common mechanism of energy coupling remains to be elucidated. Here, we discuss several models for energy‐coupling in the transport process of the transporters, largely based on currently available structures and the results of their biochemical analyses. Special attention is paid to the interaction between protonation and the negative‐inside membrane potential. Also, functional roles of the conserved sequence motifs are discussed in the context of the 3D structures. We anticipate that in the near future, a unified picture of the functions of MFS transporters will emerge from the insights gained from studies of the common architectures and conserved motifs. 相似文献
172.
While our understanding of gene-based biology has greatly improved, it is clear that the function of the genome and most diseases cannot be fully explained by genes and other regulatory elements. Genes and the genome represent distinct levels of genetic organization with their own coding systems; Genes code parts like protein and RNA, but the genome codes the structure of genetic networks, which are defined by the whole set of genes, chromosomes and their topological interactions within a cell. Accordingly, the genetic code of DNA offers limited understanding of genome functions. In this perspective, we introduce the genome theory which calls for the departure of gene-centric genomic research. To make this transition for the next phase of genomic research, it is essential to acknowledge the importance of new genome-based biological concepts and to establish new technology platforms to decode the genome beyond sequencing. 相似文献
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175.
Boon Chin Heng Yun Xia Xiaobo Shang Peter Rainer Preiser S. K. Alex Law Freddy Yin-Chiang Boey Subbu S. Venkatraman 《Biotechnology and Bioprocess Engineering》2011,16(1):127-135
Endothelial cell coverage of blood-contacting devices is crucial to their eventual success in the clinic. Two established
human cell lines derived from HUVEC (human umbilical vascular endothelial cells), CRL 2922 and CRL 2873, have been widely
utilized to study and model endothelial cell biology. However, it is not clear if these two cell lines would be useful for
modeling primary endothelial cell interaction with newly-formulated biomaterials in tissue engineering applications. Hence,
this study was conducted to compare the adhesion and proliferation characteristics of HUVEC grown on seven different substrata,
tissue culture polystyrene (TCPS), gelatin, chitosan, poly-L-lysine, hyaluronan, poly-L-lactic acid (PLLA), and polylactic-co-glycolic
acid (PLGA). The short-term adhesive behavior (2 h) of HUVEC on the various substrata was not closely-replicated by either
CRL 2873 or CRL 2922. This was likely because the 2 h timeframe is too short for identification of differences in the interaction
among the three cell types grown on various substrata. There was much faster proliferation of CRL 2922 on all seven substrata
when compared to HUVEC and CRL 2873. Moreover, the proliferation rates of CRL 2922 on the various substrata showed little
variation. In contrast, HUVEC and CRL 2873 displayed similar trends in proliferation rates, with gelatin and TCPS yielding
the highest rates, and PLLA and PLGA yielding the lowest rates. Hence, CRL 2873 is better suited for modeling primary endothelial
cell interaction with newly-formulated biomaterials than CRL 2922. The advantage of using CRL 2873 over HUVEC for biomaterial
screening is that it is immortalized and displays much less inter-batch variability than primary culture. 相似文献
176.
177.
Jordan DB Wagschal K Fan Z Yuan L Braker JD Heng C 《Journal of industrial microbiology & biotechnology》2011,38(11):1821-1835
β-d-Xylosidase/α-l-arabinofuranosidase from Selenomonas ruminantium is the most active enzyme reported for catalyzing hydrolysis of 1,4-β-d-xylooligosaccharides to d-xylose. One property that could use improvement is its relatively high affinities for d-glucose and d-xylose (K
i ~ 10 mM), which would impede its performance as a catalyst in the saccharification of lignocellulosic biomass for the production
of biofuels and other value-added products. Previously, we discovered that the W145G variant expresses K
i
d-glucose and K
i
d-xylose twofold and threefold those of the wild-type enzyme. However, in comparison to the wild type, the variant expresses 11% lower
k
cat
d-xylobiose and much lower stabilities to temperature and pH. Here, we performed saturation mutagenesis of W145 and discovered that the
variants express K
i values that are 1.5–2.7-fold (d-glucose) and 1.9–4.6-fold (d-xylose) those of wild-type enzyme. W145F, W145L, and W145Y express good stability and, respectively, 11, 6, and 1% higher
k
cat
d-xylobiose than that of the wild type. At 0.1 M d-xylobiose and 0.1 M d-xylose, kinetic parameters indicate that W145F, W145L, and W145Y catalytic activities are respectively 46, 71, and 48% greater
than that of the wild-type enzyme. 相似文献
178.
A highly selective sucrose isomerase (SIase) was purified to homogeneity from the cell-free extract of Erwinia rhapontici NX-5 with a recovery of 27.7% and a fold purification of 213.6. The purified SIase showed a high specific activity of 427.1 U mg−1 with molecular weight of 65.6 kDa. The K
m for sucrose was 222 mM while V
max was 546 U mg−1. The optimum pH and temperature for SIase activity were 6.0 and 30 °C, respectively. The purified SIase was stable in the
temperature range of 10–40 °C and retained 65% of the enzyme activity after 2 weeks’ storage at 30 °C. The SIase activity
was enhanced by Mg2+ and Mn2+, inhibited by Ca2+, Cu2+, Zn2+, and Co2+, completely inhibited by Hg2+ and Ag2+. The purified SIase was strongly inhibited by SDS, while partially inhibited by dimethylformamide, tetrahydrofuran, and PMSF.
Additionally, glucose and fructose acted as competitive inhibitors for purified SIase. 相似文献
179.
180.