https://www.massey.ac.nz/~ychisti/CellDRev.pdf

Disruption of microbial cells for intracellular products

http://enali.persiangig.com/document/PROCESS-SCALE%20DISRUPTION%20OF%20MICROORGANISMS.pdf

Process-scale disruption of microorganisms.

Formaldehyde hydrogenase and formate dehydrogenase were purified 130-fold and 19-fold respectively from Candida boidinii grown on methanol. The final enzyme preparations were homogenous as judged by acrylamide gel electrophoresis and by sedimentation in an ultracentrifuge. The molecular weights of the enzymes were determined by sedimentation equilibrium studies and calculated as 80000 and 74000 respectively. Dissociation into subunits was observed by treatment with sodium dodecylsulfate. The molecular weights of the polypeptide chains were estimated to be 40000 and 36000 respectively. The NAD-linked formaldehyde dehydrogenase specifically requires reduced glutathione for activity. Besides formaldehyde only methylglyoxal served as a substrate but no other aldehyde tested. The Km values were found to be 0.25 mM for formaldehyde, 1.2 mM for methylglyoxal, 0.09 mM for NAD and 0.13 mM for glutathione. Evidence is presented which demonstrates that the reaction product of the formaldehyde-dehydrogenase-catalyzed oxidation of formaldehyde is S-formylglutathione rather than formate. The NAD-linked formate dehydrogenase catalyzes specifically the oxidation of formate to carbon dioxide. The Km values were found to be 13 mM for formate and 0.09 mM for NAD.

https://febs.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1432-1033.1976.tb10108.x

Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii.

Mild disintegration of the green microalgae Chlorella vulgaris using bead milling

Intracellular proteins with catalytic or biological activity are of growing importance for developments in enzyme technology, as well as for the production of mammalian proteins by recombinant-DNA technology. The release of these proteins from microorganisms is an important unit operation, as it is the first step in their isolation.



Gram-scale disruption of microorganisms can be performed by a variety of established methods based on chemical, enzymatic, physical, or mechanical principles. For the large scale disruption of microorganisms, mechanical disintegrators, such as high-speed agitator bead mills or high-pressure industrial homogenizers, are commonly employed. Both types of equipment were designed originally for other tasks; in the paint industry or in the milk industry, respectively. Therefore, it appeared necessary and possible to improve design and performance for the application in cell disintegration. The goal is a uniform exposure of the microbial cells and a minimal exposure of solubilized protein to high shear forces, in order to obtain high yields and to avoid the generation of too small cell wall fragments, which are difficult to separate.



Both types of machines have been investigated for the disintegration of different microbial cells and the influence of the operating parameters analyzed on protein solubilization and enzyme yield. We will summarize the state of the art and discuss new data to illustrate trends in process development.

Purification of Proteins and the Disruption of Microbial Cells

Maltase, phosphatase, malate dehydrogenase, glucose‐6‐phosphate dehydrogenase, and 6‐phosphogluconate dehydrogenase activities were determined in the supernatant of brewer's yeast cells from samples taken at various intervals during a 90 min treatment in a Dyno‐Mill disintegrator.

Ference Marffy

Papers

Enzyme yields from cells of brewer's yeast disrupted by treatment in a horizontal disintegrator