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首页> 外文期刊>Journal of Biotechnology >Experimental pulse technique for the study of microbial kinetics incontinuous culture
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Experimental pulse technique for the study of microbial kinetics incontinuous culture

机译：用于研究微生物动力学连续培养的实验脉冲技术

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摘要

A novel technique was developed for studying the growth kinetics of microorganisms in continuous culture. The method is based on following small perturbations of a chemostat culture by on-line measurement of the dynamic response in oxygen consumption rates. A mathematical model, incorporating microbial kinetics and mass transfer between gas and liquid phases, was applied to interpret the data. Facilitating the use of very small disturbances, the technique is non-disruptive as well as fast and accurate. The technique was used to study the growth kinetics of two cultures, Methylosinus trichosporium OB3b growing on methane, both in the presence and in the absence of copper, and Burkholderia (Pseudomonas) cepacia G4 growing on phenol. Using headspace flushes, gas blocks and liquid substrate pulse experiments, estimates for limiting substrate concentrations, maximum conversion rates V-max and half saturation constants K-s could rapidly be obtained. For M. trichosporium OB3b it was found that it had a far higher affinity for methane when particulate methane monooxygenase (pMMO) was expressed than when the soluble form (sMMO) was expressed under copper limitation. While for B. cepacia G4 the oxygen consumption pattern during a phenol pulse in the chemostat indicated that phenol was transiently converted to an intermediate (4-hydroxy-2-oxovalerate), so that initially less oxygen was used per mole of phenol.
机译：开发了一种用于研究连续培养中微生物生长动力学的新技术。该方法是基于在线测量耗氧率的动态响应，跟踪化学恒温器培养物的微小扰动。结合微生物动力学和气相和液相之间的质量传递的数学模型被用来解释数据。为了便于使用很小的干扰，该技术无干扰且快速准确。该技术用于研究两种培养物的生长动力学，即在有铜和无铜下均在甲烷上生长的甲基毛孢子虫OB3b，以及在苯酚上生长的伯克霍尔德氏菌（假单胞菌）棕褐色G4。使用顶空冲洗，气体阻塞和液体基质脉冲实验，可以快速获得极限基质浓度，最大转化率V-max和半饱和常数K-s的估计值。对于曲霉孢子OB3b，发现当表达颗粒甲烷单加氧酶（pMMO）时其对甲烷的亲和力远高于在铜限制下表达可溶性形式（sMMO）时。而对于洋葱伯克霍尔德菌G4，在化学恒温器中苯酚脉冲期间的耗氧模式表明苯酚被瞬时转化为中间体（4-羟基-2-氧杂戊酸），因此最初每摩尔苯酚使用的氧气较少。

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