CELL COUNTING AND CARBON UTILIZATION VELOCITIES VIA MICROBIAL CALORIMETRY

被引:18
作者
BOE, I [1 ]
LOVRIEN, R [1 ]
机构
[1] UNIV MINNESOTA,DEPT BIOCHEM,ST PAUL,MN 55108
关键词
D O I
10.1002/bit.260350102
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Bacteria rapidly metabolize sugars and produce heat accordingly (Escherichia coli, aerobic conditions, 25°C). Two kinds of heat output are gotten: (1) from excess cells and limiting carbon, 2 × 109 to 5 × 109 cells, 5‐50 nanomole glucose; (2) from limited cells and excess carbon, 0. 1 × 109‐1 × 109 bacteria and 200‐600 nmol glucose. The thermograms from heat conduction calorimetry under the first conditions measure velocities of sugar uptake and initial metabolic throughput in 1‐6‐min time spans before a growth cycle possibly can occur. Under the second conditions with limited cells, power output plateaus to a steady state proportional to cell biomass and number of cells. In order to evaluate the calorimetric means for measuring number of cells, six independent means including spectrophotometry (turbidity) were compared: microkjeldahl nitrogen, biuret protein, dry weight, microscopy direct counting in Petroff‐Hausser chambers, and viable colony counting. Using turbidity as a central standard, all methods including calorimetry under the second set of conditions agree within ±18% of one another. Spectrophotometry is the most rapid method but is seriously interfered with by pigments that absorb and foreign particles that also scatter. Calorimetry requires 10‐30 min but measures cell numbers in opaque samples impossible for optical means. Copyright © 1990 John Wiley & Sons, Inc.
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