Combined effect of attrition and ultrasound on the disruption of Pseudomonas putida for the efficient release of arginine deiminase

Abstract

Disruption of Pseudomonas putida KT2440 by ultrasound treatment in a bath sonicator, in presence of the glass beads, was carried out for the release of arginine deiminase (ADI) and the results were compared with that of by Dyno-mill. The release of ADI depended mainly on the bead size and cellmass concentration being disrupted in bead mill. Nearly 23 U mL ADI was released when slurry with a cell-mass concentration of 250 g L was disintegrated for 9 min with 80% bead loading (0.25 mm) in Dyno-mill. Marginally higher amount of ADI (24.1 U mL ) was released by the bath sonication of 250 g L cellmass slurry for 30 min with the beads (0.1 mm) and a sonication power of 170 W. The glass beads, suspended along with the cellmass slurry in bath sonicator, efficiently disrupted the microbial cells to release ADI. Variation in the kinetic constants for the performance parameters implied that ADI release and cell disruption kinetics is a function of disruption technique used and the process variables thereof. Estimation of location factor suggested that selective release of ADI can be achieved. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1185-1194, 2018.

Oxidoreductase-Catalyzed Synthesis of Chiral Amines

“…As enzymes are produced in cells, keeping them there poses obvious advantages for biotransformations. Disruption of microbial cells and subsequent purification of the desired protein is a laborious and costly task. − The use of whole cells for biotransformations costs ca. 10% of the purified enzymes by avoiding the need for cell lysis and protein purifications. − Furthermore, implementing biocatalytic syntheses in the form of whole-cell biotransformations is advantageous in regard to in situ regeneration of the redox cofactors .…”

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

“…Despite the idea that purified protein is the most desirable form of biocatalyst, the higher costs of downstream processing limit its application on a larger scale. 510,511 Using cell free extracts that overexpress the target protein is also another popular form of biocatalysts; however, their long-term storage stability can be poor. Therefore, new techniques such as enzyme immobilization have been developed to stabilize enzymes.…”

“…Despite the idea that purified protein is the most desirable form of biocatalyst, the higher costs of downstream processing limit its application on a larger scale. , Using cell free extracts that overexpress the target protein is also another popular form of biocatalysts; however, their long-term storage stability can be poor. Therefore, new techniques such as enzyme immobilization have been developed to stabilize enzymes. − Conventional enzyme immobilization has been mediated by reactions between the reactive cAAs (Lys and Cys) of the enzyme and an activated support structure with cognate functional groups. ,, However, conventional methods usually result in randomly oriented conjugates, further reducing enzyme activity and increasing heterogeneity. − The efficiency of biocatalysis is governed by the accessibility to an intact active site, and thus the ability to construct a homogeneous array of enzymes displayed in a controlled fashion is of great interest.…”

Kinetic Resolution of Racemic Amines to Enantiopure (S)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase

“…As enzymes are intracellularly produced, keeping them in their natural environment creates apparent advantages for biocatalytic reactions [37]. The mechanical disruption of microbial cells and subsequent downstream processing for the purification of desired protein in a homogeneous form is very laborious and costly [38][39][40][41][42]. It has been reported that use of purified enzyme system for biocatalytic purposes costs approximately ten times more than that by whole-cell biotransformations [43,44].…”