ABSTRACT
BACKGROUND: LXYL-P1-2 is the first reported glycoside hydrolase that can catalyze the transformation of 7-b-xylosyl-10-deacetyltaxol (XDT) to 10-deacetyltaxol (DT) by removing the D-xylosyl group at the C7 position. Successful synthesis of paclitaxel by one-pot method combining the LXYL-P1-2 and 10- deacetylbaccatin III-10-b-O-acetyltransferase (DBAT) using XDT as a precursor, making LXYL-P1-2 a highly promising enzyme for the industrial production of paclitaxel. The aim of this study was to investigate the catalytic potential of LXYL-P1-2 stabilized on magnetic nanoparticles, the surface of which was modified by Ni2+-immobilized cross-linked Fe3O4@Histidine. RESULTS: The diameter of matrix was 2040 nm. The Km value of the immobilized LXYL-P1-2 catalyzing XDT (0.145 mM) was lower than that of the free enzyme (0.452 mM), and the kcat/Km value of immobilized enzyme (12.952 mM s 1 ) was higher than the free form (8.622 mM s 1 ). The immobilized form maintained 50% of its original activity after 15 cycles of reuse. In addition, the stability of immobilized LXYL-P1-2, maintained 84.67% of its initial activity, improved in comparison with free form after 30 d storage at 4 C. CONCLUSIONS: This investigation not only provides an effective procedure for biocatalytic production of DT, but also gives an insight into the application of magnetic material immobilization technology.
Subject(s)
Paclitaxel/biosynthesis , Glycoside Hydrolases/metabolism , Kinetics , Enzymes, Immobilized , Nanoparticles , MagnetsABSTRACT
With the rapid development of biotechnology such as NGS and proteomics , bioinformatics has seen an explo-sion in diversity and complexity in terms of data , tools and demands .The traditional computing environment , including ded-icated workstations and virtual machines , are no longer suitable under such circumstances .As a rising container technolo-gy, Docker, which is characterized by light weight , openness and security ,has provided an innovative solution to analysis and processing of biological big data and attracted increasing attention from bioinformatics developers and users .Consider-ing the demands and features of development , deployment and application of bioinformatics tools in the age of big data , this paper analyzes the advantages of Docker in this field , introduces some actual cases and discusses current deficiencies and future improvement .
ABSTRACT
The adsorption kinetics and activities of endoglucanase enzyme from Rhizopus oryzae were evaluated using activated commercial charcoal as adsorbent. The effect of various experimental parameters on adsorption of endoglucanase such as initial enzyme concentration, amount of adsorbent, contact time and temperature were investigated. The pseudo-first-order and pseudo second-order kinetic models were used to describe the kinetic data which showed that the adsorption of the enzyme followed the pseudo-first order rate expression. The adsorbed enzyme was subjected to saccharification in presence of commercially available carboxy methyl cellulose and the reusability of the adsorbed enzyme was also tested.
ABSTRACT
This study evaluated the biological importance of immobilized urease enzyme over the free urease. The support material used for urease immobilization was alginate. Generally, the immobilization of urease in alginate gel showed a marked increase in Km and Vmax. However, the immobilized urease showed higher thermal stability than that of free enzyme. The rate of thermal inactivation of the immobilized enzyme decreased due to entrapment in gel matrix. Also, the activity of the immobilized urease was more stable in retention than that of the free enzyme during the storage in solution, although the activity of the immobilized enzyme was lower in comparison with the free enzyme. A stable immobilized system and long storage life are convenient for applications that would not be feasible with a soluble enzyme system. These results highlighted the technical and biochemical benefits of immobilized urease over the free enzyme.