Analyse der biochemischen Synthesekette für Carotinoide mit Hilfe von Chlorella-Mutanten

1954 ◽  
Vol 9 (7) ◽  
pp. 461-469 ◽  
Author(s):  
Hedwig Claes

Four mutants of Chlorella vulgaris are described, which have been isolated after x-ray irradiation of the normal strain. They show blocks in the synthetic pathway of the carotenoids, along with a more or less completely blocked chlorophyll synthesis. The mixture of carotenoids formed by wild type chlorella either in light or in darkness contains mostly α- and /¿-carotene and xanthophylls (Fig. 1 a and b), whereas the mutants contain these pigments only in reduced amount or not at all. There appear instead several carotenoids of higher saturation, which are not found in wild type. Among them phytoene (Fig. 6), phytofluene (Fig. 7) and ζ-carotene (Fig. 8) were identified. Apparently they are piled up because they cannot be transformed into the ordinary end products of the synthetic chain.Mutant strain 5/871 synthesizes phytoene only. Neither phytofluene which is less saturated nor yellow pigments can be detected.The carotenoid mixture of strain 5/515 (Fig. 2) consists of phytoene, phytofluene and ζ- carotene. There is a complete block of xanthophyll synthesis.In strain 9 a xanthophylls are found (Fig. 5) but no α- or β-carotene or at least in minute amounts only. Instead the carotenoid mixture consists mostly of phytoene, phytofluene, ζ-carotene and one more carotenoid which was not yet identified. It has a maximum of absorption at 426 mμ.All mutants described so far have to be grown with glucose in darkness. They are killed by light. Another mutant strain (5/520) however was isolated which can be grown hetero- and autotrophically and shows striking differences in pigment formation depending on which condition was chosen (Fig. 3 a and 3 b). In the dark almost no chlorophyll and no xanthophylls, but phytoene, phytofluene, ζ-carotene and several unidentified carotenoids — among them possibly tetrahydrolycopene and lycopene — are produced. In the light strain 5/520 behaves like wild type with respect to chlorophyll and carotenoid synthesis. This may indicate that an enzymatically catalyzed step characteristic of wild type and missing in the mutant can be replaced here by the action of light.

1957 ◽  
Vol 12 (6) ◽  
pp. 401-407 ◽  
Author(s):  
Hedwig Claes

One of the recently described mutant strains of chlorella (strain 5/520) accumulates -in the dark phytoene, phytofluene, ζ-carotene, protetrahydrolycopene and prolycopene and forms α- and β-carotene, xanthophylls and appreciable amounts of chlorophyll in light only 1, 2.The experiments described below indicate that1. O2 and light are indispensable for xanthophyll and appreciable chlorophyll synthesis. Both syntheses appear to be linked somehow.2. Carotene synthesis depends upon light but not on O2 and is not linked to xanthophyll synthesis.3. Carotenoid synthesis in light is always accompanied by a decrease of polyenes of the Porter-Lincoln sequence, which have been accumulated in the cells during previous cultivation in the dark. Evidence seems strong that these polyenes are direct precursors of the normal carotenoids.4. An as yet unidentified carotene is formed in appreciable amounts along with the normal carotenes in the light, probably as byproduct, not a precursor.


1959 ◽  
Vol 14 (1) ◽  
pp. 4-7 ◽  
Author(s):  
Hedwig Claes

The mutant strain of Chlorella (strain 5/520) recently described synthesizes wild type carotenoids in light only. When the dark grown mutant is transferred into light, synthesis of α- and β-carotene proceeds as well in the absence as in the presence of O2, whereas synthesis of xanthophylls is dependent on light and molecular O22, 3.The experiments described below indicate that1. the final oxydative steps of the light dependent xanthophyll synthesis of Chlorella are in fact dark reactions following light reactions, the latter occurring also in N2;2. immediate precursors of xanthophylls are accumulated anaerobically in light which in the presence of molecular O2 can be oxydised to xanthophylls in the dark;3. Xanthophyll synthesis in the dark is always accompanied by a strictly corresponding decrease of α- and β-carotene accumulated in the previous light period. This would indicate that α- and β-carotene are the immediate precursors of xanthophylls in green cells.


2000 ◽  
Vol 66 (1) ◽  
pp. 64-72 ◽  
Author(s):  
Delphine Lagarde ◽  
Laurent Beuf ◽  
Wim Vermaas

ABSTRACT The psbAII locus was used as an integration platform to overexpress genes involved in carotenoid biosynthesis inSynechocystis sp. strain PCC 6803 under the control of the strong psbAII promoter. The sequences of the genes encoding the yeast isopentenyl diphosphate isomerase (ipi) and theSynechocystis β-carotene hydroxylase (crtR) and the linked Synechocystis genes coding for phytoene desaturase and phytoene synthase (crtP andcrtB, respectively) were introduced intoSynechocystis, replacing the psbAII coding sequence. Expression of ipi, crtR, andcrtP and crtB led to a large increase in the corresponding transcript levels in the mutant strains, showing that the psbAII promoter can be used to drive transcription and to overexpress various genes in Synechocystis. Overexpression of crtP and crtB led to a 50% increase in the myxoxanthophyll and zeaxanthin contents in the mutant strain, whereas the β-carotene and echinenone contents remained unchanged. Overexpression of crtR induced a 2.5-fold increase in zeaxanthin accumulation in the corresponding overexpressing mutant compared to that in the wild-type strain. In this mutant strain, zeaxanthin becomes the major pigment (more than half the total amount of carotenoid) and the β-carotene and echinenone amounts are reduced by a factor of 2. However, overexpression of ipi did not result in a change in the carotenoid content of the mutant. To further alter the carotenoid content of Synechocystis, the crtOgene, encoding β-carotene ketolase, which converts β-carotene to echinenone, was disrupted in the wild type and in the overexpressing strains so that they no longer produced echinenone. In this way, by a combination of overexpression and deletion of particular genes, the carotenoid content of cyanobacteria can be altered significantly.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Nayeong Kim ◽  
Hyo Jeong Kim ◽  
Man Hwan Oh ◽  
Se Yeon Kim ◽  
Mi Hyun Kim ◽  
...  

Abstract Background Zinc uptake-regulator (Zur)-regulated lipoprotein A (ZrlA) plays a role in bacterial fitness and overcoming antimicrobial exposure in Acinetobacter baumannii. This study further characterized the zrlA gene and its encoded protein and investigated the roles of the zrlA gene in bacterial morphology, antimicrobial susceptibility, and production of outer membrane vesicles (OMVs) in A. baumannii ATCC 17978. Results In silico and polymerase chain reaction analyses showed that the zrlA gene was conserved among A. baumannii strains with 97–100% sequence homology. Recombinant ZrlA protein exhibited a specific enzymatic activity of D-alanine-D-alanine carboxypeptidase. Wild-type A. baumannii exhibited more morphological heterogeneity than a ΔzrlA mutant strain during stationary phase. The ΔzrlA mutant strain was more susceptible to gentamicin than the wild-type strain. Sizes and protein profiles of OMVs were similar between the wild-type and ΔzrlA mutant strains, but the ΔzrlA mutant strain produced 9.7 times more OMV particles than the wild-type strain. OMVs from the ΔzrlA mutant were more cytotoxic in cultured epithelial cells than OMVs from the wild-type strain. Conclusions The present study demonstrated that A. baumannii ZrlA contributes to bacterial morphogenesis and antimicrobial resistance, but its deletion increases OMV production and OMV-mediated host cell cytotoxicity.


Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 643
Author(s):  
Konstantin Chekanov ◽  
Daniil Litvinov ◽  
Tatiana Fedorenko ◽  
Olga Chivkunova ◽  
Elena Lobakova

Carotenoids astaxanthin and β-carotene are widely used natural antioxidants. They are key components of functional food, cosmetics, drugs and animal feeding. They hold leader positions on the world carotenoid market. In current work, we characterize the new strain of the green microalga Bracteacoccus aggregatus BM5/15 and propose the method of its culturing in a bubble-column photobioreactor for simultaneous production of astaxanthin and β-carotene. Culture was monitored by light microscopy and pigment kinetics. Fatty acid profile was evaluated by tandem gas-chromatography–mass spectrometry. Pigments were obtained by the classical two-stage scheme of autotrophic cultivation. At the first, vegetative, stage biomass accumulation occurred. Maximum specific growth rate and culture productivity at this stage were 100–200 mg∙L−1∙day−1, and 0.33 day−1, respectively. At the second, inductive, stage carotenoid synthesis was promoted. Maximal carotenoid fraction in the biomass was 2.2–2.4%. Based on chromatography data, astaxanthin and β-carotene constituted 48 and 13% of total carotenoid mass, respectively. Possible pathways of astaxanthin synthesis are proposed based on carotenoid composition. Collectively, a new strain B. aggregatus BM5/15 is a potential biotechnological source of two natural antioxidants, astaxanthin and β-carotene. The results give the rise for further works on optimization of B. aggregatus cultivation on an industrial scale.


2003 ◽  
Vol 69 (10) ◽  
pp. 6114-6120 ◽  
Author(s):  
A. Hülsmann ◽  
T. M. Rosche ◽  
I.-S. Kong ◽  
H. M. Hassan ◽  
D. M. Beam ◽  
...  

ABSTRACT Vibrio vulnificus is an estuarine bacterium capable of causing rapidly fatal infections through both ingestion and wound infection. Like other opportunistic pathogens, V. vulnificus must adapt to potentially stressful environmental changes while living freely in seawater, upon colonization of the oyster gut, and upon infection of such diverse hosts as humans and eels. In order to begin to understand the ability of V. vulnificus to respond to such stresses, we examined the role of the alternate sigma factor RpoS, which is important in stress response and virulence in many pathogens. An rpoS mutant of V. vulnificus strain C7184o was constructed by homologous recombination. The mutant strain exhibited a decreased ability to survive diverse environmental stresses, including exposure to hydrogen peroxide, hyperosmolarity, and acidic conditions. The most striking difference was a high sensitivity of the mutant to hydrogen peroxide. Albuminase, caseinase, and elastase activity were detected in the wild type but not in the mutant strain, and an additional two hydrolytic activities (collagenase and gelatinase) were reduced in the mutant strain compared to the wild type. Additionally, the motility of the rpoS mutant was severely diminished. Overall, these studies suggest that rpoS in V. vulnificus is important for adaptation to environmental changes and may have a role in virulence.


2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Mamta Singh ◽  
Prabhakar Tiwari ◽  
Garima Arora ◽  
Sakshi Agarwal ◽  
Saqib Kidwai ◽  
...  

Abstract Inorganic polyphosphate (PolyP) plays an essential role in microbial stress adaptation, virulence and drug tolerance. The genome of Mycobacterium tuberculosis encodes for two polyphosphate kinases (PPK-1, Rv2984 and PPK-2, Rv3232c) and polyphosphatases (ppx-1, Rv0496 and ppx-2, Rv1026) for maintenance of intracellular PolyP levels. Microbial polyphosphate kinases constitute a molecular mechanism, whereby microorganisms utilize PolyP as phosphate donor for synthesis of ATP. In the present study we have constructed ppk-2 mutant strain of M. tuberculosis and demonstrate that PPK-2 enzyme contributes to its ability to cause disease in guinea pigs. We observed that ppk-2 mutant strain infected guinea pigs had significantly reduced bacterial loads and tissue pathology in comparison to wild type infected guinea pigs at later stages of infection. We also report that in comparison to the wild type strain, ppk-2 mutant strain was more tolerant to isoniazid and impaired for survival in THP-1 macrophages. In the present study we have standardized a luciferase based assay system to identify chemical scaffolds that are non-cytotoxic and inhibit M. tuberculosis PPK-2 enzyme. To the best of our knowledge this is the first study demonstrating feasibility of high throughput screening to obtain small molecule PPK-2 inhibitors.


1988 ◽  
Vol 47 (3) ◽  
pp. 425-431 ◽  
Author(s):  
GERALD F. DEITZER ◽  
BENJAMIN A. Horwitz ◽  
Jonathan Gressel

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