Gamma-aminobutyric acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography.

We have developed a gamma-aminobutyric acid (GABA) production technique using his-tag mediated immobilization of Escherichia coli-derived glutamate decarboxylase (GAD), an enzyme that catalyzes the conversion of glutamate to GABA. The GAD was obtained at 1.43 g/L from GAD-overexpressed E. coli fermentation and consisted of 59.7% monomer, 29.2% dimer and 2.3% tetramer with a 97.6% soluble form of the total GAD. The harvested GAD was immobilized to metal affinity gel with an immobilization yield of 92%. Based on an investigation of specific enzyme activity and reaction characteristics, glutamic acid (GA) was chosen over monosodium glutamate (MSG) as a substrate for immobilized GAD, resulting in conversion of 2.17 M GABA in a 1 L reactor within 100 min. The immobilized enzymes retained 58.1% of their initial activities after ten consecutive uses. By using cation exchange chromatography followed by enzymatic conversion, GABA was separated from the residual substrate and leached GAD. As a consequence, the glutamic acid was mostly removed with no detectable GAD, while 91.2% of GABA was yielded in the purification step.

QoS improvement with an optimum controller selection for software-defined networks.

The software-defined networking (SDN) paradigm has simplified the management of computer networks by decoupling data and control planes. Moreover, the separation of the data and control planes has transitioned network complexity from traditional devices to controllers; therefore, controllers have become indispensable entities in SDN. Controllers have multiple features and direct the network from a central point and respond to updates to topological changes. However, the supportive capability of these features is strong in one controller but weak in another. Due to several controllers and each controller having a set of features, selecting an optimal SDN controller can be considered to be a multi-criteria decision-making (MCDM) problem. Herein, a two-step approach is proposed for SDN controller selection. First, the controllers are ranked with analytical network process (ANP) according to their qualitative features which influence the performance of these controllers and then a performance comparison is performed to check for the QoS improvement. The controller with a high-weight value from the feature-based comparison is quantitatively analysed by experimental analysis. The main contribution of this paper is checking the applicability of the ANP for controller selection in SDN considering its features and performance analysis in real-world Internet and Brite topologies. The simulation results show that the controller computed through the proposed approach outperforms the controller selected with existing approaches. The selection of an optimum controller with ANP results in a reduction of topology discovery time and delay in the normal and traffic load scenario. Similarly, an increase in throughput with a reasonable utilization of the central processing unit (CPU) is observed for the proposed controller.

High-level conversion of L-lysine into 5-aminovalerate that can be used for nylon 6,5 synthesis.

L-Lysine is a potential feedstock for the production of bio-based precursors for engineering plastics. In this study, we developed a microbial process for high-level conversion of L-lysine into 5-aminovalerate (5AVA) that can be used as a monomer in nylon 6,5 synthesis. Recombinant Escherichia coli WL3110 strain expressing Pseudomonas putida delta-aminovaleramidase (DavA) and lysine 2-monooxygenase (DavB) was grown to high density in fed-batch culture and used as a whole cell catalyst. High-density E. coli WL3110 expressing DavAB, grown to an optical density at 600 nm (OD600 ) of 30, yielded 36.51 g/L 5AVA from 60 g/L L-lysine in 24 h. Doubling the cell density of E. coli WL3110 improved the conversion yield to 47.96 g/L 5AVA from 60 g/L of L-lysine in 24 h. 5AVA production was further improved by doubling the L-lysine concentration from 60 to 120 g/L. The highest 5AVA titer (90.59 g/L; molar yield 0.942) was obtained from 120 g/L L-lysine by E. coli WL3110 cells grown to OD600 of 60. Finally, nylon 6,5 was synthesized by bulk polymerization of ϵ-caprolactam and δ-valerolactam prepared from microbially synthesized 5AVA. The hybrid system demonstrated here has promising possibilities for application in the development of industrial bio-nylon production processes.