Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method

Imada, Chiaki; Ikemoto, Yuko; Kobayashi, Takeshi; Hamada, Naoko; Watanabe, Etsuo

doi:10.1046/j.1444-2906.2002.00438.x

Cited by 7 publications

(2 citation statements)

References 10 publications

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“…But, as is already known, ethanol is easy to evaporate. According to the literature, the liquid method of protoplast regeneration is an attractive process to greatly increase the frequency of protoplast regeneration compared with the agar method [ 20 ]. To facilitate the precision of genome shuffling procedures, in our experiment, we used liquid RM to regenerate protoplast to prevent ethanol volatilization.…”

Section: Discussionmentioning

confidence: 99%

Direct Ethanol Production from Lignocellulosic Sugars and Sugarcane Bagasse by a RecombinantTrichoderma reeseiStrain HJ48

Chen

Wei

Wang

et al. 2014

The Scientific World Journal

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Trichoderma reesei can be considered as a candidate for consolidated bioprocessing (CBP) microorganism. However, its ethanol yield needs to be improved significantly. Here the ethanol production of T. reesei CICC 40360 was improved by genome shuffling while simultaneously enhancing the ethanol resistance. The initial mutant population was generated by nitrosoguanidine treatment of the spores, and an improved population producing more than fivefold ethanol than wild type was obtained by genome shuffling. The results show that the shuffled strain HJ48 can efficiently convert lignocellulosic sugars to ethanol under aerobic conditions. Furthermore, it was able to produce ethanol directly from sugarcane bagasse, demonstrating that the shuffled strain HJ48 is a suitable microorganism for consolidated bioprocessing.

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Section: Discussionmentioning

confidence: 99%

Direct Ethanol Production from Lignocellulosic Sugars and Sugarcane Bagasse by a RecombinantTrichoderma reeseiStrain HJ48

Chen

Wei

Wang

et al. 2014

The Scientific World Journal

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“…They studied amylase activity of fusants and found 2.5 % higher than that of parental strains. Imada et al (2002) proposed a simple and quicker procedure for protoplast fusion.…”

Section: Protoplast Fusionmentioning

confidence: 99%

Preparation and fusion of protoplasts from the phytopathogenic fungus Sclerotium rolfsii (Sacc.)

Sikandar¹

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1.11. Epidemiology 1.12. Control of Disease 1.13. Aims of Thesis 2. Materials and Methods 2.1. Organisms and Growth conditions 2.2. Preparation of semi-permeable cellophane membrane 2.3. Pretreatment of mycelia 2.4. Digestion mixtures 2.4.1. Osmotic stabilizers 2.4.2. Lytic enzymes 2.5. Protoplast formation 2.6. Determination of protoplast number 2.7. Purification of protoplast 2.8. Determination of protoplast size 2.9. Regeneration of protoplasts 2.9.1. Regeneration on solid medium 2.9.2. Regeneration in liquid medium 2.10. Fusion of protoplast 2.11. Study of stability of fusants 2.12. Growth rate of fusants 2.13. Pathogenic properties of fusants in green house pot experiment 2.13.1. Inoculum preparation 2.13.2. Soil preparation 2.13.3. Selecetion of host crops 66 2.13.4. Seed soaking and planting 66 2.13.5. Study of growth parameters of tomato, carrot and beans 67 2.13.5.1 Growth parameters studied 67 2.13.6. Disease incidence and mortality 68 2.13.7. Chlorophyll fluorescence 68 2.13.8. Estimation of chlorophyll a, b and carotenoids 69 2.14. Statistical analysis 70 3. Results 71 3.1.Strains A and R of Sclerotium rolfsii 71 3.2. Optimization of protoplast yields from Sclerotium rolfsii 71 3.2.1. Effect of pretreatment of mycelia 71 3.2.2. Effect of the osmotic stabilizers on protoplast yields and morphology74 3.2.2.1. Effect of different molarities of MgSO 4 on protoplast yields 76 3.2.3. Effect of different lytic enzymes on protoplast yields 76 3.3. Measurement of size of protoplast 3.4. Mechanism of protoplast release from the hyphae of Sclerotium rolfsii 3.5. Regeneration of protoplast into hyphae 3.6. Fusion of protoplasts of strains A and R 3.6.1. Recovery of the fusants 88 3.6.2. Morphological characters of fusants 3.6.3. Stability of the fusants 92 3.6.4. Growth rate of fusants 92 3.7. Assessment of pathogenicity of fusants and parent strains A and R of Sclerotium roflii 94 3.8. Physiological parameters 3.8.1.Chlorophyll fluorescence 3.8.2. Estimation of chlorophyll a, b and carotenoids 4. Discussion 4.1. Investigation into optimization for protoplast yields 4.3. Protoplast release mechanism 4.4. Protoplast regeneration 4.5. Protoplast fusion 4.6. Properties of fusants 4.7. Pathogenicity of fusants 5. References

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Evolution of Streptomyces pristinaespiralis for resistance and production of pristinamycin by genome shuffling

Jin

Wang

et al. 2008

Appl Microbiol Biotechnol

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Improvement of pristinamycin production by Streptomyces pristinaespiralis was performed by using recursive protoplast fusion and selection for improved resistance to the product antibiotic in a genome shuffling format. A 100-microg/ml pristinamycin resistant recombinant, G 4-17, was obtained after four rounds of protoplast fusion, and its production of pristinamycin reached 0.89 g/l, which was increased by 89.4% and 145.9% in comparison with that of the highest parent strain M-156 and the original strain CGMCC 0957, respectively. The subculture experiments indicated that the hereditary character of high producing S. pristinaespiralis G 4-17 was stable. It is concluded that genome shuffling improves the production of pristinamycin by enhancing product-resistance in a stepwise manner. Pristinamycin fermentation experiments by recombinant G 4-17 were carried out in a 5-l fermentor, and its production of pristinamycin reached 0.90 g/l after 60 h of fermentation.

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Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method

Cited by 7 publications

References 10 publications

Direct Ethanol Production from Lignocellulosic Sugars and Sugarcane Bagasse by a RecombinantTrichoderma reeseiStrain HJ48

Direct Ethanol Production from Lignocellulosic Sugars and Sugarcane Bagasse by a RecombinantTrichoderma reeseiStrain HJ48

Preparation and fusion of protoplasts from the phytopathogenic fungus Sclerotium rolfsii (Sacc.)

Evolution of Streptomyces pristinaespiralis for resistance and production of pristinamycin by genome shuffling

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