By Mary E. Lidstrom (Eds.)
This quantity is the 1st complete compilation of equipment for the assay and purification of enzymes fascinated with the usage of diminished one-carbon compounds and better hydrocarbons. within the final decade there was an explosion of curiosity within the organisms that develop on one-carbon compounds and in those who develop on larger hydrocarbons. this can be as a result of the advertisement curiosity within the certain enzymes enthusiastic about those area of expertise metabolic pathways and to a starting to be knowing of the $64000 position those organisms play in carbon biking in nature. more moderen curiosity within the use of those organisms and their enzymes for detoxing of detrimental waste has once more positioned the highlight on those detailed microorganisms. This quantity will end up to be a worthwhile resource for researchers in academia, executive, and alike
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A. DUINE Alcohol + dye ~ aldehyde + dye "H2 Pseudomonas species involved in the degradation of alkanes contain NAP(P)+-independent alcohol dehydrogenases, but the nature of the cofactor is unknown. 2 P. putida a,4 (the organism originally supposed to be Acinetobacter calcoaceticus 4 is now classified as a P. putida strain), and Comamonas testosteroni (P. testosteroniS). Although these are quinoproteins [enzymes with pyrroloquinoline quinone (PQQ) as a cofactor], two quite different types of enzymes appear to exist, namely, quinoprotein and quinohemoprotein alcohol dehydrogenase.
Protein concentrations are determined by the method of Lowry et al. 7 Purification Procedure Growth of the Organism. 53 is grown at 35 ° on a mineral medium. 5 g MgC12" 6H20, and 3 g (NH4)2SO4. After autoclaving, 1 ml calcium solution, 1 ml iron solution, and 1 ml trace element solution (all sterilized by autoclaving) are added per liter of basic medium. 5% (v/v) ethanol (sterilized by filtration). Cultivation is carried out in a 2-liter flask with reciprocal shaking (250 rpm). 0. The cell paste is stored at - 4 0 °' Purification Steps.
Acetone or acetol monooxygenase activity, corrected for endogenous activity, is expressed as nanomoles 02 consumed per minute per milligram protein. The dissolved oxygen concentration of air-saturated buffer is calculated using the method of Robinson and Cooper. 6 It is worth 5 S. Hartmans and J. A. M. deBont, FEMSMicrobiolLett. 36, 151 (1986). 6 j. Robinson and R. M. Cooper, Anal. Biochem. 33, 390 (1970).  PROPANE UTILIZATION ENZYME ASSAYS 31 noting that no acetone monooxygenase activity can be detected in Rhodococcus rhodochrous PNKbl, despite the use of different buffer systems and substitution of NADPH with a variety of other electron donors.