Flow cytometry for the intracellular pH measurement of glutamate producing Corynebacterium glutamicum

Leyval, Damien; Debay, Florence; Engasser, Jean‐Marc; Goergen, Jean‐Louis

doi:10.1016/s0167-7012(97)00031-6

Cited by 22 publications

(10 citation statements)

References 20 publications

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“…The Xuorescence was proportional to the pH i of the cells grown in fermenters between 6.6 and 8.3 [52]. In this study, pH i was found to be a good indicator of the physiological state of the cells, and FCM a sensitive and rapid tool for measuring cytoplasmic pH.…”

Section: Intracellular Phmentioning

confidence: 58%

Flow cytometry applications in the food industry

Comas-Riu

Rius

2009

J Ind Microbiol Biotechnol

101

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Flow cytometry has become a valuable tool in food microbiology. By analysing large numbers of cells individually using light-scattering and fluorescence measurements, this technique reveals both cellular characteristics and the levels of cellular components. Flow cytometry has been developed to rapidly enumerate microorganisms; to distinguish between viable, metabolically active and dead cells, which is of great importance in food development and food spoilage; and to detect specific pathogenic microorganisms by conjugating antibodies with fluorochromes, which is of great use in the food industry. In addition, high-speed multiparametric data acquisition, analysis and cell sorting, which allow other characteristics of individual cells to be studied, have increased the interest of food microbiologists in this technique. This mini-review gives an overview of the principles of flow cytometry and examples of the application of this technique in the food industry.

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Section: Intracellular Phmentioning

confidence: 58%

Flow cytometry applications in the food industry

Comas-Riu

Rius

2009

J Ind Microbiol Biotechnol

101

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“…The pH optimum for the conversion of L-lactate to pyruvate is higher (between 8.0 and 8.5), but there is still considerable activity at neutral pH owing to a shoulder in the profile. The intracellular pH of C. glutamicum has been measured to vary between pH 7.7-8.3 during batch-and fed-batch culture (Leyval et al 1997), a pH range compatible with a CgLDH reverse activity.…”

Section: Discussionmentioning

confidence: 99%

Kinetic characterisation of recombinant Corynebacterium glutamicum NAD+-dependent LDH over-expressed in E. coli and its rescue of an lldD − phenotype in C. glutamicum: the issue of reversibility re-examined

et al. 2011

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The ldh gene of Corynebacterium glutamicum ATCC 13032 (gene symbol cg3219, encoding a 314 residue NAD+-dependent L-(+)-lactate dehydrogenase, EC 1.1.1.27) was cloned into the expression vector pKK388-1 and over-expressed in an ldhA-null E. coli TG1 strain upon isopropyl-β-D-thiogalactopyranoside (IPTG) induction. The recombinant protein (referred to here as CgLDH) was purified by a combination of dye-ligand and ion-exchange chromatography. Though active in its absence, CgLDH activity is enhanced 17- to 20-fold in the presence of the allosteric activator D-fructose-1,6-bisphosphate (Fru-1,6-P2). Contrary to a previous report, CgLDH has readily measurable reaction rates in both directions, with Vmax for the reduction of pyruvate being approximately tenfold that of the value for L-lactate oxidation at pH 7.5. No deviation from Michaelis-Menten kinetics was observed in the presence of Fru-1,6-P2, while a sigmoidal response (indicative of positive cooperativity) was seen towards L-lactate without Fru-1,6-P2. Strikingly, when introduced into an lldD- strain of C. glutamicum, constitutively expressed CgLDH enables the organism to grow on L-lactate as the sole carbon source.

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“…Recently, flow cytometry has been used to quantify the response of C. glutamicum to antibiotics [191] and to measure intracellular pH as well as transmembrane potential [192]. This kind of monitoring of single cell properties might, in future, improve process monitoring and control especially because changes in the composition of a bacterial population might even be detected at an early stage of fermentation so that early decisions to counteract them can be made.…”

Section: Single Cell Analysis By Flow Cytometrymentioning

confidence: 99%

Microbial Production of l-Amino Acids

Faurie¹,

Thommel²,

Bathe³

2002

Advances in Biochemical Engineering/Biotechnology

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Advances in Biochemical Engineering/Biotechnology reviews actual trends in modern biotechnology. Its aim is to cover all aspects of this interdisciplinary technology where knowledge, methods and expertise are required for chemistry, biochemistry, microbiology, genetics, chemical engineering and computer science. Special volumes are dedicated to selected topics which focus on new biotechnological products and new processes for their synthesis and purification. They give the state-of-the-art of a topic in a comprehensive way thus being a valuable source for the next 3-5 years. It also discusses new discoveries and applications.In general, special volumes are edited by well known guest editors. The managing editor and publisher wilt however always be pleased to receive suggestions and supplementary information. Manuscripts are accepted in English.In references Advances in Biochemical Engineering/Biotechnology is abbreviated asAdv Biochem Engin/Biotechnot as a journal.A complete list of all enzymes entries either as an alphabetical Name Index or as the EC-Number Index is available at the above mentioned URL. You can download and print them free of charge.A complete list of all synonyms (more than 25,000 entries) used for the enyzmes is available in print form, ISBN 3-540-41830-X. Save 15 %We recommend a standing order for the series to ensure you automatically receive all volumes and all supplements and save 15% on the list price.Preface XI Due to the comparably high prices, L-tryptophan, which is an important pharmaceutical active ingredient produced by Ajinomoto and Amino GmbH, has been launched in the feed market in smaller quantities so far.Ajinomoto however announced in 2001 the intention to increase the annual rate to more than 1,000 tons. This is a very good example of the close interdependence between the cost of production, the target price and the amount of amino acid consumed in order to reduce primary feed stuff for animal farming.Roughly estimated, there is an inverse proportional linear relationship between the logarithm of the market price and the annual production of each amino acid. This observation forces each amino acid manufacturer to construct more efficient bacterial strains and to optimize corresponding fermentation protocols allowing a maximal specific product-and space-time-yield in order to minimize variable and fixed costs of production. This has become possible by application of the modern techniques offered by modern molecular biology and biotechnology, including the use of recombination combined with metabolic flux analysis and functional genomics.The authors of this special volume of "Advances in Biochemical Engineering/Biotechnology" have contributed significantly to the progress of amino acid biotechnology in the last decades and earn our special gratitude and admiration for their expert review articles.In order to understand the impact of the economical aspect of amino acid production we have rounded off this volume with an article dealing with this topic.Frellstedt,

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Flow cytometry for the intracellular pH measurement of glutamate producing Corynebacterium glutamicum

Cited by 22 publications

References 20 publications

Flow cytometry applications in the food industry

Flow cytometry applications in the food industry

Kinetic characterisation of recombinant Corynebacterium glutamicum NAD+-dependent LDH over-expressed in E. coli and its rescue of an lldD − phenotype in C. glutamicum: the issue of reversibility re-examined

Microbial Production of l-Amino Acids

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