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Carbon as a Parameter in Bacterial Systems Growth Limitation and Substrate Utilization Studies M. D. RICKARD, Research Engineer Chemicals Laboratory W. H. RILEY, Biochemist Biochemical Research Laboratory 1701 Building The Dow Chemical Company Midland, Michigan INTRODUCTION The need for more definitive parameters for following bacterial growth and substrate utilization has led the authors to the application of infrared carbon analysis as a parameter for the study of the behavior of bacterial systems. The method is based on the rapid combustion of a micro sample in oxygen and measurement of the CO2 produced with a nondispersive infrared analyzer (1). Although designed primarily for the analysis of the carbon content of waste waters (2), the technique has application to the study of bacterial systems since it makes possible a rapid, accurate determination of tne organic carbon content of biological samples in solution or suspension. The object of this paper is to survey the utility of carbon analysis in following the metabolism of organic compounds during the growth of a bacterial culture in a defined medium. The relationships obtained among cellular carbon, exogenous soluble carbon, and viable count during bacterial growth are shown along with the rates of synthesis of cellular material. MATERIALS AND METHODS Culture Methods Aerobacter aerogenes ATCC No. 8308 was used throughout these studies. The organism was cultured in a glucose-mineral salts medium in an incubated shaker at 30 C. The composition of the medium used is shown in Table I. The stock culture was maintained on nutrient agar slants and was passed through three serial transfers in the medium before the cells were used. Experimental Treatment Cells from a 24-hr culture were harvested by centrifugation and washed with sterile phosphate buffer (M/15 - pH 7.0). These cells were then suspended in fresh sterile medium for use as an inoculant. The experiments were performed in a 16-liter fermenter which was maintained in a 30 C water bath. Aeration and agitation of the contents were accomplished through the use of an air diffuser placed in the bottom of the vessel. Filtered, compressed air was passed through the diffuser at a sufficient rate to insure adequate aeration and mixing of the culture during respiration. The fermenter's contents were sampled through the use of a positive siphon. - 98 -
Object Description
Purdue Identification Number | ETRIWC196509 |
Title | Carbon as a parameter in bacterial systems growth limitation and substrate utilization studies |
Author |
Rickard, M. D. Riley, W. H. |
Date of Original | 1965 |
Conference Title | Proceedings of the twentieth Industrial Waste Conference |
Conference Front Matter (copy and paste) | http://earchives.lib.purdue.edu/u?/engext,12162 |
Extent of Original | p. 98-109 |
Series |
Engineering extension series no. 118 Engineering bulletin v. 49, no. 4 |
Collection Title | Engineering Technical Reports Collection, Purdue University |
Repository | Purdue University Libraries |
Rights Statement | Digital object copyright Purdue University. All rights reserved. |
Language | eng |
Type (DCMI) | text |
Format | JP2 |
Date Digitized | 2009-05-19 |
Capture Device | Fujitsu fi-5650C |
Capture Details | ScandAll 21 |
Resolution | 300 ppi |
Color Depth | 8 bit |
Description
Title | page 98 |
Collection Title | Engineering Technical Reports Collection, Purdue University |
Repository | Purdue University Libraries |
Rights Statement | Digital object copyright Purdue University. All rights reserved. |
Language | eng |
Type (DCMI) | text |
Format | JP2 |
Capture Device | Fujitsu fi-5650C |
Capture Details | ScandAll 21 |
Transcript | Carbon as a Parameter in Bacterial Systems Growth Limitation and Substrate Utilization Studies M. D. RICKARD, Research Engineer Chemicals Laboratory W. H. RILEY, Biochemist Biochemical Research Laboratory 1701 Building The Dow Chemical Company Midland, Michigan INTRODUCTION The need for more definitive parameters for following bacterial growth and substrate utilization has led the authors to the application of infrared carbon analysis as a parameter for the study of the behavior of bacterial systems. The method is based on the rapid combustion of a micro sample in oxygen and measurement of the CO2 produced with a nondispersive infrared analyzer (1). Although designed primarily for the analysis of the carbon content of waste waters (2), the technique has application to the study of bacterial systems since it makes possible a rapid, accurate determination of tne organic carbon content of biological samples in solution or suspension. The object of this paper is to survey the utility of carbon analysis in following the metabolism of organic compounds during the growth of a bacterial culture in a defined medium. The relationships obtained among cellular carbon, exogenous soluble carbon, and viable count during bacterial growth are shown along with the rates of synthesis of cellular material. MATERIALS AND METHODS Culture Methods Aerobacter aerogenes ATCC No. 8308 was used throughout these studies. The organism was cultured in a glucose-mineral salts medium in an incubated shaker at 30 C. The composition of the medium used is shown in Table I. The stock culture was maintained on nutrient agar slants and was passed through three serial transfers in the medium before the cells were used. Experimental Treatment Cells from a 24-hr culture were harvested by centrifugation and washed with sterile phosphate buffer (M/15 - pH 7.0). These cells were then suspended in fresh sterile medium for use as an inoculant. The experiments were performed in a 16-liter fermenter which was maintained in a 30 C water bath. Aeration and agitation of the contents were accomplished through the use of an air diffuser placed in the bottom of the vessel. Filtered, compressed air was passed through the diffuser at a sufficient rate to insure adequate aeration and mixing of the culture during respiration. The fermenter's contents were sampled through the use of a positive siphon. - 98 - |
Resolution | 300 ppi |
Color Depth | 8 bit |
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